PRODUCTION OF 50 MTPA POLYHYDROXYBUTYRATE FROM

JATROPHA OIL

by

NURUL AIN BT IBRAHIM

QASTALANI BT GHAZALI
SHOBANA A/P SINNIAH
NUR FATIN NADIAH BT FAUZI

KE12004
KE11004
KE11058
KE11042

A design project submitted to the Faculty of Chemical and Natural Resources

Engineering in partial fulfillment of the requirements for the degree of
Bachelor of Chemical Engineering (Biotechnology)

Faculty of Chemical and Natural Resources Engineering

Universiti Malaysia Pahang

DECEMBER 2014

Universiti Malaysia Pahang

Faculty of Chemical and Natural Resources Engineering

The undersigned certify that they have read, and recommend to the Faculty of
Chemical and Natural Resources Engineering for acceptance, a design project
entitled Production of 50 MTPA Polyhydroxybutyrate from Jatropha Oil
submitted by
NURUL AIN BT IBRAHIM
QASTALANI BT GHAZALI
SHOBANA A/P SINNIAH
NUR FATIN NADIAH BT FAUZI

KE12004
KE11004
KE11058
KE11042

in partial fulfillment of the requirements for the degree of Bachelor of

Chemical Engineering in Biotechnology.

Dr. Nur Hidayah Bt Mat Yasin

Date:

ACKNOWLEDGEMENT

First and foremost, we would like to express our greatest gratitude and
sincere appreciation to our plant design supervisor, Dr. Nur Hidayah Binti Mat
Yassin for his exemplary guidance, monitoring and constant encouragement
throughout the process to complete this Plant Design Project. We appreciated
all efforts of supervisor in advising and be available at right time besides
providing valuable insights leading to the successful completion of our plant
design project. Without her guidance and help of overseeing the whole
progress of the team works until the end, we would not be able to accomplish
this design project successfully.
Besides, we would like to take this opportunity to express a deep sense
of gratitude to Dr Mior Ahmad Khusairi Bin Mohd Zahari and Mr. Rozaimi
Abu Samah for their cordial support, valuable information and guidance,
which helped us in completing this task through various stages especially in
simulation.
Furthermore, we are obliged to thank all panels during 3 stages of plant
design presentations for the valuable comments and information provided by
them in their respective fields. All these useful comments help a lot in
improving our plant design project. In addition, sincere thankful is also
extended to our lecturers who had provided us with assistance and
encouragements at any occasions. In addition, we would like to thank our
parents for their unconditional love in giving us support and motivation which
enable us to be determined and without giving up in completing the plant
design project.
Last but not least, to our beloved course mates and acquaintance,
constant encouragement and exchange of knowledge throughout our struggles
in completing this design project. May this report will benefits all readers not
only us in designing new plant for production of Polyhydroxybutyrate (PHB)
for overall stages.

ii
EXECUTIVE SUMMARY

Litter is a problem with a very negative social and environmental

impact. One way to tackle this problem is to use biodegradable plastics as an
environmentally-friendly solution for things such as plastic
bags. Biodegradable plastics are plastics that can be broken down by
microorganisms (bacteria or fungi) into water, carbon dioxide (CO2) and some
bio-material. Polyhydroxybutyrate is a polymer belonging to the polyesters
class that are of interest as bio-derived and biodegradable plastics.
Therefore the objective of this plant and process design is to develop a
new (PHB) plant using Jatropha oil as the main carbon source and
Cupriavidus necator H16 as the biomass or the microorganism. Urea is
selected as nitrogen source as it could produce high PHB content. According
to a new study by World Bioplastics from The Freedonia Group. Inc, it stated
that global demand for biodegradable and bio-plastics will be more than triple
to more than 1.1 million tons in 2015, valued at $2.9 billion. Demand for
biodegradable polyesters is said to be growing by about 27.9% for a five years.
This is due to customer demand for more environmentally-sustainable
products, development of bio-based feed stocks for commodity plastic resins,
increasing restrictions on the use of non-degradable plastic products and high
rise of crude oil and natural gas prices. Frost & Sullivan have examined
current and future of the bioplastics market in Southeast Asia for the period
2004 to 2014. It stated that the bioplastics market is at a developing stage. The
total market for engineering plastics in Southeast Asia in 2007 was 12 tons.
These units are forecast to grow at a rate of about 129.8 percent per year and
reach about 4063 tons by 2014 (Sullivan, 2008). Malaysias first fully
automated PHA Bioplastics Pilot Plant was launched and scaled-up to 2,000 L,
the bioreactor facilities and integrated manufacturing process of the plant are
able to produce various options of PHA materials from crude palm kernel oil
and palm oil mill effluent. Bioplastics based on PHA in 2013 has been
projected to reach 0.5 billion kg. According to observation of market survey, it
is proposed to produce 50 metric ton PHB per annum. The location is decided
to produce 50 MTPA of PHB which is at Sungai Bako area Kuching, Sarawak.

iii
Plant layout which consists of administration building, operational building,
waste treatment plant, laboratory and research center, and other ancillary
buildings has been sketched.
Based on the calculation, the total power usage in plant is 719.008
kWh. Average industrial tariff for electricity from Sarawak is 33.70 sen /kWh.
By applying the industrial tariff of electricity 33.70 sen /kWh, the total
electricity cost per year is equal to RM 4797652.781/year with operation hours
of 8000 per year. All the calculation is based on CEPCI 2014. The total steam
usages for main equipment are 1,186.67kg/h. Based on calculation using the
standard steam charges, the total steam cost is about RM 9140191.175 /year
with operation hours of 8000 per year. By conversion, the total steam cost is
RM 9140191.175 /year. The total water consumption for bioreactors and seed
fermenters is 12,849.51 kg/batch. Through calculation the total cost of water is
RM 7454515.37/year with operation hours 8000 per year. RM
7454515.37/year is needed for water cost. By addition of total cost by
electricity, steam and water cost, the total cost of utilities is RM 4797652.781
+ RM 9140191.175 + RM 7454515.37 = RM 21392359.33 /year.
This plant consists of five major equipment. There are seed fermenter,
fermenter, blending storage, disk stack centrifuge and spray dryer. Each of the
equipment has their own hazard. Hazard identification procedure is used to
identify the types of adverse health effects that can be caused by exposure to
some agent in question, and to characterize the quality and weight of evidence
supporting this identification. Risk assessment includes determination of the
events that can produce an accident, the probability of those events, and
consequences that could include human injury or loss of life, damage to the
environment, or loss of production and capital equipment. Hazard
identification can be performed independent of risk assessment, but it would
obtain best result if they are done together.
Economic and profitability analysis in the form of discounted cash
flow will be evaluated in this report as an effort to estimate profit or loss of
this PHB plant. Grass root capital (GRC) is the cost of equipment installation
in a plant and it costs major portion of total fixed capital cost. From

iv
calculation, it is determined that GRC for this PHB plant is approximately
RM3, 570,000.00. While as for the total capital investment (TCI) for this PHB
plant is approximately RM5, 378, 000.00. Profitability analysis will be
determined in this report by evaluating operating margin. Operating margin is
the ratio of operating profit to sales and it indicates how much of each
Malaysian Ringgit is left after operating expenses. A high operating margin
means that the plant has good cost control and that sales are increasing faster
than costs.

TABLE OF CONTENTS
ACKNOWLEDGEMENT .................................................................................. i
EXECUTIVE SUMMARY ............................................................................... ii
TABLE OF CONTENTS ................................................................................... v
LIST OF FIGURES ........................................................................................viii
LIST OF TABLES ............................................................................................ ix
CHAPTER 1 ...................................................................................................... 1
INTRODUCTION ............................................................................................. 1
1.1
Background ......................................................................................... 1
1.1.1
Plastics ......................................................................................... 1
1.1.2
Biodegradable plastics ................................................................. 2
1.1.3
Poly-(3-hydroxybutyrate), PHB ................................................... 2
1.1.4
Physical and chemical properties of PHB .................................... 3
1.1.5
Biodegradability of PHB.............................................................. 4
1.1.6
Storage and Handling ................................................................... 5
1.2
Applications of PHB ........................................................................... 5
1.2.1
Medical ........................................................................................ 6
1.2.2
Aquaculture .................................................................................. 6
1.2.3
Pharmaceutical ............................................................................. 7
1.3
Market Survey ..................................................................................... 7
1.3.1
Global Market Demand................................................................ 7
1.3.2
Asian market demand .................................................................. 8
1.3.3
Malaysia market demand ............................................................. 8
1.3.4
Global production ........................................................................ 9
1.3.5
Future Prospect of PHB ............................................................. 10
1.3.6
Prices of Products, Raw Materials and Chemicals .................... 11
1.3.7
Jatropha Oil ................................................................................ 12
1.4
Screening of Synthesis Routes .......................................................... 15
1.4.1
Synthesis routes for PHB production ......................................... 15
1.4.2
Selected synthesis route ............................................................. 24
1.4.3
Utilization of Jatropha oil .......................................................... 24
1.4.4
Type of Microbial Production Strain ......................................... 24
1.4.5
Feeding source of nutrient supply .............................................. 26
1.4.6
PHB synthesis ............................................................................ 26
1.4.7
Downstream Process .................................................................. 27
CHAPTER 2 .................................................................................................... 28
PROCESS FLOW SHEETING ....................................................................... 28
2.1
Selection of raw material and impurities management ..................... 28
2.2
Input and Output Flow Sheeting ....................................................... 28
2.2.1
Mechanical Equipment Description ........................................... 33
2.3
Material and Energy Balances ........................................................... 35
2.3.1
Material Balance ........................................................................ 36
2.3.2
Energy Balance .......................................................................... 64
2.4
Economic Potential ........................................................................... 71
2.4.1
Economic Potential 2 Based On Input and Output Structure .... 71
2.4.2
Economic Potential 3 Based On Recycle Structure .................. 75
2.5
Comparison of Simulation (SuperPro) and Manual Calculation
Results .......................................................................................................... 78

vi
CHAPTER 3.................................................................................................. 79
UTILITIES & HEAT INTERGRATION ........................................................ 79
3.1
Introduction ....................................................................................... 79
3.2
Utilities .............................................................................................. 79
3.2.1
Electricity ................................................................................... 79
3.2.2
Steam.......................................................................................... 80
3.3
Heat Integration ................................................................................. 81
3.4
Economic Potential Level 5: Heat Integration System ..................... 84
CHAPTER 4 .................................................................................................... 85
EQUIPMENT SIZING .................................................................................... 85
4.1
Introduction ....................................................................................... 85
4.2
Heat Sterilizer (ST-101 & ST-102) ................................................... 85
4.3
Media Preparation Tank (P-09) ......................................................... 86
4.4
Splitter (FSP-101 & FSP-102) .......................................................... 86
4.5
Gas Compressor (G-101)................................................................... 87
4.6
Air Filter (AF-101 & AF-102) .......................................................... 87
4.7
Seed Fermenter (V-101) .................................................................... 87
4.8
Main Fermenter (V-103) ................................................................... 89
4.9
Storage Tank (V-104)........................................................................ 90
4.10 Centrifuge (DS-101, DS-102 & DS-103) .......................................... 90
4.11 Pumps ................................................................................................ 91
4.12 Spray Dryer (SDR-101) .................................................................... 92
4.13 Economic Potential Level 4 (EP4): Separation System .................... 92
4.13.1 General Structure of the Separation System .............................. 92
CHAPTER 5 .................................................................................................... 94
PROCESS CONTROL & SAFETY ................................................................ 94
5.1
Introduction ....................................................................................... 94
5.2
Identification of Hazard..................................................................... 94
5.2.1
Material Safety Data Sheet ........................................................ 95
5.2.2
DOW Fire and Explosion Index ................................................ 97
5.2.2
Toxicity .................................................................................... 104
5.3
Hazard and Operability Studies (HAZOP) of Major Equipment .... 106
5.4
Major Equipment Control ............................................................... 108
5.4.1
Objectives of Control System .................................................. 109
5.4.2
Process Control of Major Equipment....................................... 110
5.5
Piping and Instrumentation Diagram .............................................. 113
CHAPTER 6 .................................................................................................. 114
WASTE MANAGEMENT AND POLUTION CONTROL ......................... 114
6.1
Introduction ..................................................................................... 114
6.1.1
Higher Up the Hierarchy .......................................................... 115
6.1.2
Waste Minimization ................................................................. 116
6.1.3
Objective of Waste Minimization ............................................ 117
6.1.4
Waste Sources and Effect to Human and Environment ........... 117
6.1.5
Waste Management Option for Each Waste Produced ............ 118
6.2
JABATAN ALAM SEKITAR (JAS) Schedule B and EQA
ENVIRONMETAL QUALITY ACT, 1974 .............................................. 122
6.2.1
Gaseous Emission .................................................................... 122
6.2.2
Sewage, Industrial Effluent and Leachate Discharge .............. 125
6.3
Waste Treatment Option ................................................................. 128
6.3.1
Biological Method ................................................................... 128

vii
6.3.2
Chemical Method ..................................................................... 129
6.3.3
Physical Method....................................................................... 130
6.3.4
Selection of Method ................................................................. 130
6.4
Process Description ......................................................................... 131
6.5
Waste in Polyhydroxybutyrate (PHB) Plant ................................... 132
CHAPTER 7 .................................................................................................. 135
SITE SELECTION AND PLANT LAYOUT ............................................... 135
7.1
Introduction ......................................................................................... 135
7.2
General Consideration of Plant Location ........................................ 135
7.3
Type of Industry Preferred and Location ........................................ 136
7.3.1
Availability of Raw Material ................................................... 136
7.3.2
Utilities ..................................................................................... 137
7.3.3
Water Supply ........................................................................... 137
7.3.4
Electricity Supply..................................................................... 138
7.3.5
Land Selling Price and Area Still Available ............................ 138
7.3.6
Transportation System ............................................................. 139
7.3.7
Availability of Manpower ........................................................ 140
7.3.8
Research and Development Organization ................................ 140
7.3.9
Geography, Climate and Environment .................................... 140
7.3.10 Government Incentive .............................................................. 141
7.3.11 Waste and Effluent Disposal Facilities .................................... 141
7.4
Site Selection analysis ..................................................................... 141
7.5
Plant Layout .................................................................................... 142
7.5.1
Introduction .............................................................................. 142
7.5.2
Definition ................................................................................. 142
7.5.3
Objectives of Plant Layout....................................................... 143
7.5.4
Factors Affecting the Plant Layout .......................................... 144
CHAPTER 8 .................................................................................................. 150
ECONOMIC ANALYSIS ............................................................................. 150
8.1
Introduction ..................................................................................... 150
8.2
Grass Root Capital .......................................................................... 150
8.3
Capital Investment........................................................................... 152
8.4
Manufacturing Cost ......................................................................... 153
8.5
Cash Flow Analysis ......................................................................... 159
8.5.1
Payback Period Analysis.......................................................... 159
8.6
Profitability Analysis....................................................................... 165
8.7
Conclusion ....................................................................................... 166
CHAPTER 9 .................................................................................................. 167
9.1
Conclusion ....................................................................................... 167
9.2
Recommendation ............................................................................. 168
REFERENCES ................................................................................................. vi
APPENDICES .................................................................................................. vi

viii

LIST OF FIGURES
Figure 1. 1: Chemical Structure of Polyhydroxybutyrate .................................. 3
Figure 1. 2: Global Production capacities of bioplastics in 2012 (by region) ... 9
Figure 1. 3: World Biodiesel Production, 2005-2017 (Millions of gallons) in
Indonesia, Argentina, Brazil, U.S, and Europe. ............................................... 10
Figure 1. 4: TEM of Cupriavidus necator showing PHB inclusion bodies ..... 14
Figure 1. 5: Total acreage of jatropha oil plantations in selected countries
(Extracted from: http://www.jatropha-alliance.com) ....................................... 20
Figure 2. 1: Block Flow Diagram of PHB Production..................................... 30
Figure 2. 2: Block Flow Diagram of Upstream Process .................................. 31
Figure 2. 3: Block Flow Diagram from Downstream Process ......................... 32
Figure 2. 4: Process Flow Diagram of PHB plant ........................................... 33
Figure 2. 5: Process Flow Diagram Simulation in SuperPro ........................... 34
Figure 2. 6: Input-output structure of PHB production process....................... 71
Figure 2. 7: Graph of concentration versus conversion of Jatropha oil ........... 73
Figure 2. 8: Diagram of Recycle ...................................................................... 75
Figure 2. 9: Graph of product, biomass, recycled biomass, Jatropha oil, and
urea concentration versus Jatropha oil conversion. ......................................... 76
Figure 2. 10: Graph of economic potential at the second level (EP2), economic
potential at the third level with recycle and economic potential at the third
level without recycle. ....................................................................................... 78
Figure 5. 1: Procedure of hazard identification and risk assessment. (Source:
Guidelines for Hazards Evaluation Procedures: American Institute of
Chemical Engineers, 1985) .............................................................................. 95
Figure 5. 2: General steps in determining DOW Fire and Explosion Index .... 99
Figure 5. 3: Form used in DOW Fire and Explosion Index ........................... 100
Figure 5. 4: Section of P & ID of Seed Fermenter......................................... 111
Figure 5. 5: Section of P & ID of Main Fermenter ........................................ 112
Figure 5. 6: Section of P & ID of Disc Stack Centrifuges ............................. 112
Figure 5. 7: Section of P & ID of Spray Dryer .............................................. 113
Figure 6. 1: Waste management hierarchy .................................................... 114
Figure 6. 2: Conceptual Flow Diagram for Activated Sludge Wastewater
Treatment System .......................................................................................... 131
Figure 7. 1: Plant Layout of PHB plant ......................................................... 146
Figure 8. 1: Undiscounted Cash Flow............................................................ 161
Figure 8. 2: Discounted Cash Flow................................................................ 163

ix

LIST OF TABLES
Table 1. 1: Chemical properties of PHB ............................................................ 4
Table 1. 2: World Bioplastic Demand for 2005 2015 ..................................... 7
Table 1. 3: Media for the production of PHB .................................................. 11
Table 1. 4: The prices of the material components .......................................... 11
Table 1. 5: Properties of Jatropha oil from Bionas Sdn. Bhd. ......................... 13
Table 1. 6: Characteristics of urea ................................................................... 14
Table 1. 7: Raw materials and their prices for production of 50 MTPA PHB
using soybean oil as carbon source. ................................................................. 17
Table 1. 8: Raw materials and their prices for production of 50 MTPA PHB
using crude palm kernel oil (CPKO) as carbon source. ................................... 18
Table 1. 9: Raw materials and their prices for production of 50 MTPA PHB
using Jatropha oil as carbon source. ................................................................ 21
Table 1. 10: Comparison between soybean oil, jatropha oil, and crude palm
kernel oil based on its availability of raw materials, yield of PHB, concerns,
cost and operation mode. ................................................................................. 22
Table 1. 11: comparison between promising microorganisms in PHB
cultivation, an analysis from Choi and Lee (1997). ......................................... 25
Table 2. 1: Input and Output of Heat Sterilizer (ST-101&ST-102) ................. 36
Table 2. 2: Density of Each Components ........................................................ 38
Table 2. 3: Summary of materials used in blending tank................................. 42
Table 2. 4: Amount of input of Seed Fermenter and Main Fermenter ............ 43
Table 2. 5: Input and Output of Compressor (G-101) ..................................... 43
Table 2. 6: Input and Output of Air Filter (AF-101)........................................ 44
Table 2. 7: Summary of material used in seed fermenter ................................ 47
Table 2. 8: Summary of materials used in main fermenter .............................. 48
Table 2. 9: Summary amount of input into Seed Fermenter and Main
Fermenter ......................................................................................................... 54
Table 2. 10: Summary amount of output from fermenter ................................ 55
Table 2. 11: Overall material balance of Seed Fermenter................................ 55
Table 2. 12: Summary of Overall Material Balance of main fermenter .......... 56
Table 2. 13: Input and output of Air Filter (AF-102) ...................................... 57
Table 2. 14: Input and Output of Flat Bottom Tank (V-104) .......................... 57
Table 2. 15: Input and output of Centrifugal (C-101) ...................................... 58
Table 2. 16: Input and Output of Blending Tank (C-101) ............................... 59
Table 2. 17: Input and output streams of mixer (MX-101).............................. 59
Table 2. 18: Input and Output Stream of Disc-stack Centrifuge (P-13/DS-102)
.......................................................................................................................... 61
Table 2. 19: Input and Output Stream of Blending Tank (P-14/V-103) .......... 62
Table 2. 20: Input and output of Disc-stack Centrifugal (C-03) ...................... 62
Table 2. 21: Summary Input Stream of Spray Dryer (P-16/SDR-101)............ 63
Table 2. 22: Summary Output of Spray Dryer (P-16/SDR-101) ..................... 63
Table 2. 23: Summary of energy balance of each stream ................................ 65
Table 2. 24: Heat of formation ......................................................................... 68
Table 2. 25: Heat duty for each equipment ...................................................... 70
Table 2. 26: Values for EP2 calculation .......................................................... 74

x
Table 2. 27: Graph of EP2 versus Jatropha oil conversion .............................. 74
Table 2.27: Table 2. 28: Data for EP2 and EP3 at both with recycle and
without recycle. ................................................................................................ 77
Table 2. 29: Comparison between Simulation and Manual Balance ............... 78
Table 3. 1: Total Power consumption of equipment used in plant design ....... 79
Table 3. 2: Total steam consumption of equipment used in plant design ........ 80
Table 3. 3: Total water consumption of equipment used in plant design ........ 81
Table 4. 1: Sizing Summary of Heat Sterilizer ................................................ 85
Table 4. 2: Sizing Summary of Media Preparation Tank ................................ 86
Table 4. 3: Sizing Summary of Splitter............................................................ 86
Table 4. 4: Sizing Summary of Gas Compressor ............................................. 87
Table 4. 5: Sizing Summary of Air Filter ........................................................ 87
Table 4. 6: Bare Module Cost (CBM) for Centrifuges .................................... 93
Table 5. 1: Degree of Hazard based on DOW Fire and Explosion Index (FEI)
........................................................................................................................ 103
Table 5. 2: Toxicity level ............................................................................... 104
Table 5. 3: Toxicity rating system ................................................................. 105
Table 5. 4: General Guide Words for HAZOP procedures (Crowl and Louvar,
2002) .............................................................................................................. 106
Table 6. 1: Source and Waste Generated in PHB plant ................................. 117
Table 6. 2: Waste Management Options by Our Company ........................... 118
Table 6. 3: Malaysian Standard Guidelines for Air Gaseous Pollutants ........ 119
Table 6. 4: Malaysia, Canada and USA Ambient Air Quality Guidelines .... 120
Table 6. 5: Characterization of Waste Type According to MIDA ................. 121
Table 6. 6: Comparison of Aerobic and Anaerobic Treatment (Mittal, 2011)
........................................................................................................................ 128
Table 6. 7: Total Gaseous Waste ................................................................... 133
Table 6. 8: Total Waste Summary ................................................................. 133
Table 6. 9: Costing for Waste Treatment Option Employed in Our Company
........................................................................................................................ 134
Table 7. 1: Water Provider Based on Location .............................................. 137
Table 7. 2: Electricity Provider Based On Location ...................................... 138
Table 7. 3: Building and Location in the Plant Layout .................................. 147
Table 8. 1: Bare Module Cost of Equipment in PHB Plant ........................... 150
Table 8. 2: Estimation of Grass Root Capital, GRC. ..................................... 152
Table 8. 3: Fixed and Total Capital Investment ............................................. 153
Table 8. 4: Estimation of Operating Labor Cost ............................................ 155
Table 8. 5: Summary of Manufacturing Cost ................................................ 156
Table 8. 6: Cash Flow Analysis for Undiscounted Rate, I% ......................... 160
Table 8. 7: Discounted Cash Flow Summary ................................................ 162

xi
Table 8. 8: Net Present Value for Discounted Rate ....................................... 164
Table 8. 9: Discounted Cash Flow at DCFRR=28.35% ................................ 165

CHAPTER 1

INTRODUCTION

1.1

Background
This chapter will provides overview of Polyhydroxybutyrate (PHB) as

well as other components involved in the plant. The demand and supply of
PHB also was discussed.

1.1.1

Plastics
Plastics are man- made long chain polymeric molecules similar in

many ways to natural resins found in trees and other plants (Scott, 1999). On
the other hand, plastics are uniquely exible materials that have seen them
occupy a huge range of functions, from simple packing materials to complex
engineering components (Jim and Alexandre et al., 2013). The history of
plastic begins from 1862 by Alexander Parkes. The main raw material in
plastic production is petroleum. The properties of plastic which is high
molecular weight and tightly bonded together make the plastic not degradable,
their disposal become difficult and give negative impact on the environment
(Sharmila et al., 2011). During the 1980s, the solid waste problem emerged as
a potential crisis in many areas of the US because of increasing amounts of
municipal solid waste (MSW), shrinking landfill capacity, rising costs and
strong public opposition to new solid waste facility sittings (Regan et al.,
1990). In 1960 plastics made waste less than half a percent of US MSW
generation. By 2010 they made up to 12.4% and only 8.2% is recovered (US
EPA, 2011).

2
1.1.2

Biodegradable plastics
Biodegradable plastics were introduced in the 1980s to find ways to

produce non-petroleum based plastics as well as to reduce the environmental

effects because of the increased landfill (Gironi and Piemonte, 2010).
According to European Bioplastics, a plastic material is defined as a bioplastic
if it is either biobased, biodegradable, or features both properties. The term
biobased means that the part of material or product is derived from biomass.
Meanwhile, biodegradation is a chemical process which could be degraded by
the microorganism in the environment when proper conditions such as the
sunlight, moisture, oxygen and so forth are available convert materials into
natural substances such as water, carbon dioxide, and composition (Abe and
Doi, 2002).

1.1.3

Poly-(3-hydroxybutyrate), PHB
Polyhydroxybutyrate (PHB) is a polyhydroxyalkanoate (PHA), a

polymer belong to the polyesters class that was first isolated and characterized
in 1925 by French microbiologist Maurice Lemoigne. PHB is produced by
microorganisms (like Ralstonia eutropha or recombinant Escherichia coli)
apparently in response to conditions of physiological stress. The polymer is
primarily a product of carbon assimilation (from glucose or starch) and is
employed by microorganisms as a form of energy storage molecule to be
metabolized when other common energy sources are not available. Microbial
biosynthesis of PHB starts with the condensation of two molecules of acetylCoA

to

give

acetoacetyl-CoA

which

is

subsequently

reduced

to

hydroxybutyryl-CoA. This latter compound is then used as a monomer to

polymerize PHB (Lemoigne, 2009).
Since 1925, PHB has been produced through bacterial fermentation
(Rosa, 2004), being synthesized under limited culture conditions, and it is
usually produced through the use of microorganisms that belong to genres
Alcaligenes, Azobacter, Bacillus, and Pseudomonas (Ugur, 2002).

3
The poly-3-hydroxybutyrate (P3HB) form of PHB is probably the most
common type of polyhydroxyalkanoate, but many other polymers of this
class are produced by a variety of organisms: these include poly-4hydroxybutyrate (P4HB), polyhydroxyvalerate (PHV), polyhydroxyhexanoate
(PHH), polyhydroxyoctanoate (PHO) and their copolymers (Lemoigne, 2009).
Poly-(3-hydroxybutyrate), PHB is one of the most important members
of PHAs. According to Li et al. (1999), PHB is an intracellular carbon and
energy storage material produced by many microorganisms under unfavorable
growth condition such as limitation of (NH4)2SO4, PO32-, Mg2+ and oxygen.
PHB is synthesized from acetyl-CoA using three enzymatic steps (Paramjit
and Nitika, 2011). It is a biodegradable thermoplastic polyester which can be
used in various ways like the conventional non-degradable plastics (Li et al.,
1999). The chemical structure of PHB is shown as in Figure 1.1.

Figure 1. 1: Chemical Structure of Polyhydroxybutyrate

1.1.4

Physical and chemical properties of PHB

The physical properties of PHB are elastomeric, insoluble in water,

nontoxic, biocompatible, and piezoelectric, with high degree of polymerization

(Samantary et al, 2011). Besides, PHB is also resistant to water and ultraviolet
radiation and impermeable to oxygen. In addition, PHB is a partially
crystalline material with high melting temperature and high degree of
crystallinity. PHB is stiff and brittle. PHB does not contain any residues of
catalyst and is perfectly isotactic and does not include any chain branching. It
is not water soluble but is 100% biodegradable. PHB has low permeable for
O2, H2O and CO2 (Samantary et al, 2011). Chemical properties of PHB is
summarized in Table 1.1.

4
Table 1. 1: Chemical properties of PHB
Parameter

Value

Melting point (oC)

171-182

Glass transition temperature (oC)

5-10

Crystallinity (%)

65-80

Density (g cm-3)

1.23 - 1.25

Molecular weight (g/mol)

6600000

Molecular weight distribution

2.2 3

Heat capacity (kJ/kg.K)

1.465

Tensile strength [MPa]

40

Extension to break [%]

68

UV resistance

Good

Solvent resistance

Poor

Oxygen permeability [cm3m-2atm-1d-1]

45

Biodegradability

Good

1.1.5

Biodegradability of PHB
Biodegradation of PHB is dependent upon a number of factors such as

the microbial activity of the environment and the exposed surface area. In
addition, temperature, pH, molecular weight and crystallinity are important
factors. Biodegradation starts when microorganisms begin growing on the
surface of the plastic and secrete enzymes that break down the polymer into its
molecular building blocks, called hydroxyacids. The hydroxyacids are then
taken up by the microorganisms and used as carbon sources for growth. In
aerobic environments the polymers are degraded to carbon dioxide and water.
The environmental degradation behavior of PHB-g-VAc films (Xg:
0%, 5% and 15%) before and after saponification assessed by the BOD
method in environmental water. Many kinds of PVA-utilizing microorganisms have been found in the water of major rivers (Matsumura et al.,
1994), and it was confirmed that PVA could be degraded in environmental
water from the lake at the Takasaki Advanced Radiation Research Institute.

5
Biodegradation of PHA has also been tested in various aquatic
environments. In one study in Lake Lugano, Switzerland, items were placed at
different depths of water as well as on the sediment surface. A life span of 510 years was calculated for bottles under these conditions (assuming no
increase in surface area), while PHA films were completely degraded in the
top 20 cm of sediment within 254 days at temperatures not exceeding 6C.

1.1.6

Storage and Handling

PHB is non-toxic biopolymer. Therefore, it is biocompatible and hence

is suitable for medical applications. It is important to minimize premature

PHB degradation during fabrication and storage. This is because PHB are
biodegradable polymer and its biodegradation is dependent upon a number of
factors such as the microbial activity of the environment and the exposed
surface area. In addition temperature, pH, molecular weight and crystallinity
are also play an important role. In one report, the maximum biodegradation
rates were observed at moisture level of 55% and temperatures of around
60C. Therefore, it is well advised to packed PHB in airtight, aluminumbacked, or plastic foil pouches and kept it in the refrigerator.

1.2

Applications of PHB
There are many applications of PHB besides it is been used in the

production of biodegradable plastic. PHB have been chosen as petroleum

derived plastic replacement because of its properties that possess high
durability and endurance similar like regular plastics but unlike regular
plastics, it can be decomposed to water and carbon dioxide aerobic
microorganisms existing from sewage, sea or soil without forming any toxic
products. PHB can be used as wrapping materials like bags, containers and
throwaway items such as cup, plates and diapers.

6
1.2.1

Medical
Since biodegradability and biocompatibility are properties of PHB, the

combination of PHB with hydroxyapatite (HA) were used as scaffolding

material in tissue engineering (Brigham, 2012). For instance, the medical
practitioners use PHB scaffolding material to treat bone defects. While the
combinations of copolymer of polyglycolic acid (PGA) and PHB was used to
produce pulmonary valve leaflets and pulmonary artery scaffolds in sheep.
PHB also used in medical devices such as for dental and skin surgery
(Bonartsev, 2007). The efficiency of these devices in term of biocompatibility,
biodegradation and therapeutic is still in progress.

1.2.2

Aquaculture
PHB was used as a food add-on to aquaculture animals in order to

control the enormous deaths caused by pathogenic contaminations. Larvae of

the brine shrimp Artemia franciscana serve as important feed in fish and
shellfish larviculture however, they are subject to bacterial diseases that
devastate entire populations and consequently hinder their use in aquaculture.
It was found that PHB might shield the fish meal which is gnotobiotic brine
shrimp Artemia franciscana against pathogenic vibriosis (Schryver, 2010).
The release of the PHB monomer -hydroxybutyric acid was suggested to
inhibit the growth and/or the activity of the pathogens (Schryver, 2010). By
integrating the accumulation of PHB in bio-flocs, this technique can possibly
decrease the rate of death during larval and young stages of aquaculture
animals and can therefore become all the more cost effective. No adverse
effects were observed when the feed is introduced for about 10% to the diet of
the fish.

7
1.2.3

Pharmaceutical
In pharmaceutical, PHB is applied into slow-released carrier for lasting

drug delivery due to their biocompatibility and biodegradability properties. It

is also used as cell and tablet packaging material. PHB, 3-hydroxyhexanoate
(PHBHHx), and polylactic acid (PLA) were used to study drug sustained
release. The results showed that over a period of at least 20 days for PHB and
PHBHHx nanoparticles, while PLA nanoparticles and free drug lasted only 15
days and a week, respectively (Xiong, 2010).

1.3

Market Survey

1.3.1

Global Market Demand

Polyhydroxybutyrate (PHB) and similar polymers have obtained

worldwide interest because of their biodegradability in addition to their

durability and plasticity. Industrial production of PHA and other
biodegradable plastics is shown in Table 1.2.
Table 1. 2: World Bioplastic Demand for 2005 2015
WORLD BIOPLASTICS DEMAND
(thousand metric tons)
Item

2005 2010 2015

%Annual Growth
2005 - 2010

2010-2015

Bioplastics Demand

130

300

1025

18.2

27.9

North America

34

80

242

18.7

24.8

Western Europe

60

125

347

15.8

22.7

Asia/Pacific

33

83

320

20.3

31.0

Other Regions

12

116

32.0

57.4

According to a new study, World Bioplastics, from The Freedonia

Group, Inc; it stated that global demand for biodegradable and bio-plastics will

8
more than triple to more than 1 million metric tons (1.1 million tons) in 2015,
valued at $2.9 billion. Demand for biodegradable polyesters is said to be
growing by about 27.9% for a five years, and North America is belatedly
catching up with other regions for about 24.8 % of annual growth for 2010 to
2015. This is due to customer demand for more environmentally-sustainable
products, development of bio-based feed stocks for commodity plastic resins,
increasing restrictions on the use of non-degradable plastic products and high
rise of crude oil and natural gas prices.

1.3.2

Asian market demand

Frost & Sullivan have examined the bioplastics markets in Southeast

Asia. The research service presents current and future of the bioplastics market
in Southeast Asia for the period 2004 to 2014. It stated that the bioplastics
market is at a developing stage. The total market for engineering plastics in
Southeast Asia in 2007 was 12 tons. These units are forecast to grow at a rate
of about 129.8 percent per year and reach about 4063 tons by 2014 (Sullivan,
2008).

1.3.3

Malaysia market demand

Malaysias first fully automated PHA Bioplastics Pilot Plant was

launched by Science, Technology and Innovation Minister Datuk Seri Dr.

Maximus Johnity Ongkili at Jalan Beremban. Scaled-up to 2,000 L, the
bioreactor facilities and integrated manufacturing process of the plant are able
to produce various options of PHA materials from crude palm kernel oil and
palm oil mill effluent. Bioplastics based on PHA in 2013 has been projected to
reach 0.5 billion kg (First-Of-Its-Kind Sirim Bioplastics Pilot Plant Launched
in 2011).

9
1.3.4

Global production

Figure 1. 2: Global Production capacities of bioplastics in 2012 (by region)

Bioplastics production capacities are growing fastest outside of

Europe. In 2012 production capacities amounted to approximately 1.4 million
tons. Market data of European Bioplastics forecasts production capacities will
multiply by 2017 to more than 6 million tons. Based on the figure above, it
has shown that Asia has dominated the production of bioplastics which is 36.2
percent. It is about 0.5 million tons per year (Bioplastics, 2014).

10

Figure 1. 3: World Biodiesel Production, 2005-2017 (Millions of gallons)

in Indonesia, Argentina, Brazil, U.S, and Europe.

The market of around 1.2 million tons in 2011 may see a five-fold
increase in production volumes by 2016, to almost 6 million tons. The product
expected to contribute most to this growth is bio-based PET (for plastic
bottles), which already accounts for approximately 40% of the global
bioplastics production capacity. The current production volume is expected to
grow to more than 4.6 million tons by 2016 as a result of demand from large
manufacturers of carbonated drinks. Early in 2013 the nova-Institute predicted
that by 2020 bioplastics production could rise to 12 million tons, principally
due to drop-in polymers, particularly bio-PET13. With an expected total
polymer production of about 400 million tons in 2020, the bio-based share
should increase from 1.5% in 2011 to 3% in 2020 (Development, 2013).

1.3.5

Future Prospect of PHB

Polyhydroxybutyrate (PHB) is diverse and versatile class of materials

that has potential applications in many sectors of the economy. Currently,

11
productions of PHB are still in the developmental stage, but important
applications are beginning to emerge in packaging, food production, and
medicine. We have reached a critical point in the development of PHBs for
many applications. It is, therefore, an opportune time for a comprehensive
report detailing promising new developments in this field. In brief, production
of PHB has good future prospect because:
1. PHB is biodegradable
2. Production of PHB protects the fossil resources
3. PHBs have a positive eco-balance sheet
4. Good example for a sustainable development in the spirit of the
agenda of 21st century
5. Carbon dioxide neutral
6. The use of biodegradable material creates over 20, 000 new and
secure workplaces in Europe and many times over in the world
(social factor)

1.3.6

Prices of Products, Raw Materials and Chemicals

Types of raw materials and amount used for pre-cultures are shown in

Table 1.3 while the market prices for PHB, raw materials of culture medium in
PHB production are shown in Table 1.4:
Table 1. 3: Media for the production of PHB
Materials

Amount (g/L)

Jatropha oil

20.00 g/L

Urea

1.00 g/L

Table 1. 4: The prices of the material components

Materials

Prices (MYR/kg)

Source

Jatropha oil

2.73

Bionas Malaysia Sdn Bhd

12

1.3.7

Urea

1.70

Petronas Fertilizer Sdn Bhd

PHB

27.45

Jatropha Oil
Jatropha oil is a potential renewable resource because Jatropha

plantations yield large amounts of oil, are highly resistant to drought and pests
and the oil is relatively cheap and non-edible. Jatropha oil is derived from
Jatropha curcas seeds. This plant was originally found in the Caribbean area
but is now widespread throughout Africa, the Americas and much of Asia. The
plant also is known as hardy Jatropha due to its resistance to pest and
drought, and also its ability to grow almost anywhere. The oil yield of this
plant is almost four times that of soybean, and 10 times that of maize.
Recently, Jatropha oil has been evaluated as a source of high quality biodiesel
production. However, it has not been evaluated as a feedstock for PHA
production (Ko-Sin Ng, 2010).
The genus Jatropha belongs to the Euphorbiaceous family which can
synthesize several toxic compounds, including carcinogenic phorbol ester,
trypsin inhibitor, lectin and saponin. The toxins render the oil non-edible, but
should not affect its utility for bioplastics production. In view of the above, it
is advantageous to use Jatropha oil which is not food-grade oil as the sole
carbon source to produce PHA (Ko-Sin Ng, 2010).
It has three Malaysian entities and six overseas joint ventures which
are Bionas Murabahah Bhd, Bionas Sdn Bhd and Biofuel Bionas Sdn Bhd,
Bionas Philippines, Bionas Indonesia, Bionas Vietnam, Bionas Cambodia,
Bionas Thailand and Bionas Taiwan. Its assets portfolio consists of over
600,000 acres planted areas, 3.3 million acres land bank, 313 seedling
nurseries & harvest collection centers and 3 processing plants.
As a result, the company has monthly supply and production capacity
of 100,000 tons seeds, 90,000 tons seedlings, 33,000 MT Crude Jatropha Oil
(CJO) and 65,000 MT seed cakes (bio-mass). Now the company is extending

13
its global presence by expanding to Taiwan, Thailand, Cambodia, Bangladesh
and Cambodia. With Bionas diverse operational experience and an unrivaled
business heritage, it is now poised to take a leading position in the global
business arena.
In the year 2008 and 2009, Bionas has been actively promoting the
cultivation of Jatropha in Malaysia. The numbers of planters had risen from
28,983 in 2008 to 112,484 in 2009 respectively. Bionas has also increased the
numbers of nurseries from 98 in 2008 to 221 in 2009. The number of planters
had risen to 238,541 and the number of nurseries to 313 in 2010. The company
has also setup four pressing mills in 2010 as part of the companys capacity
building to cater its needs for the production Bionas Jatropha Additives. 2011
has been a productive period for Bionas as the company has invested into the
setting up of two processing, blending plant, and storage facilities for Bionas
Jatropha Additives at two main ports of Malaysia which are located in Prai
Port, Penang and Kuching Port, Sarawak.

Table 1. 5: Properties of Jatropha oil from Bionas Sdn. Bhd.

Criteria

Properties

Climate type

Tropical
37%

Seed oil content

st

rd

Average annual yield/Acre (1 -3

year)

3.6 MT

Lifespan

50 years

Harvest period

Monthly after six months

Crude oil price (MYR/MT)

2736.00

By-products

Seed cakes i.e Biomass Briquette

1.3.7.3 Biomass: Cupriavidus Necator

14
Cupriavidus necator was formerly known as Alcaligenes eutrophus is a
motile, rod shaped, Gram negative, non-sporing bacterium and major strains is
H16 and JMP 134 (Larsen and Pogliano, 2007). Larsen and Pogliano (2011)
stated that its optimal temperature is 30C while optimal pH is 7 and it is a
non-halophilic, which cannot live in high salt concentration. It is able to
produce PHB inside the inclusion bodies under limited nitrogen source but
excessive carbon source (Ojumu et al. 2004).

Figure 1. 4: TEM of Cupriavidus necator showing PHB inclusion bodies

1.3.7.4 Urea
Urea is a white crystalline substance with the chemical formula CO
(NH2)2. It is highly water soluble and contains 46% nitrogen. Urea is
considered an organic compound because it contains carbon. It was the first
organic compound ever synthesized by chemists; this was accomplished in the
early 1800s. Urea supplies more nitrogen per ton of product than any other dry
fertilizer. It contains 46% nitrogen; this means that each ton of urea supplies
920 lbs. of nitrogen.
Table 1. 6: Characteristics of urea

15
Other Name

Urea

Molecular Formula

CH4N2O

Molecular weight

60.06

1.4

Screening of Synthesis Routes

PHB has a great potential as a biodegradable bio plastic. However, the

major drawback to the commercialization of PHB is their high cost of

production compared with conventional petrochemical based plastic materials.
The cost of carbon feed stocks or raw materials required can significantly
affect the PHB production cost in large production scale. Therefore, the
production cost can be considerably lowered when alternative cost-effective
carbon feedstock, type of microbial production strain, as well as nutrient
supply during the biosynthesis for the commercialization of bio plastics are
identified.

1.4.1

Synthesis routes for PHB production

Various substrates especially plant oils have been evaluated as an

excellent carbon source in PHA production. Examples given are soybean oil
(Kahar et al., 2004), palm oil (Kek et al., 2008) and Jatropha oil (Khan et al.,
2013). Kahar et al. (2008) reported that plant oils are desirable as they are also
inexpensive carbon sources. Additionally, due to their high carbon content,
plant oils yield almost two-fold higher than from glucose and they are
appealing feed stocks for industrial PHA production because metabolism of
these compounds can influence the monomer composition of the resulting
PHA (Akiyama et al., 2003).

1.4.1.1 Production of PHB from soybean oil by Cupriavidus necator H16

16
According to Kahar et al. (2008), high yield production of
polyhydroxyalkanoates has been identified from using soybean oil by wildtype strain Ralstonia eutropha or now known as Cupriavidus necator, one of
the best known bacteria among PHA-producing microorganisms. Soybean oil
has a high yield of PHA obtained ranging from 0.72 to 0.76 g PHA/ g soybean
oil used and the PHA productivity obtained here was roughly calculated to be
1.0 g/L.h (Kahar et al. 2008).
According to Global Agricultural Information Network (GAIN) in
2012, there is no commercial cultivation of soybeans in Malaysia despite it is
one of the largest producers of soy drinks in Southeast Asia, with exports
going to neighboring countries as well as Australia, Japan and Europe.
Malaysia has to import soybean oil from U.S, hence this will increase
production cost as shipping and handling cost have to be considered.
Furthermore, contrary to sugar that can be directly utilized by cells,
soybean oil needs to be hydrolyzed by lipase and fatty acids. This would
increase production cost to acquire lipase and fatty acids as well as equipment
such as hydrolyzer. Environmental wise, large scale production from soybean
oil is environmentally friendly as the carbon dioxide emission from soybean
oil are very low compared to the petrochemical polymers if high yield of PHA
is produced (Kahar et al. 2008). However, the use of edible oils in production
of bio plastics may cause depletion of global food supply and sources. Using
soybean oil is considered as unethical as it is wasteful to convert food to bio
plastics. Additionally, Ng et al. (2010) also stated that the edible plant oils
price has increased drastically because of recent crisis of food shortage and
increase of food demand.

1.4.1.1.1

Economic Potential Level 1

As we produce 50,000 kg of PHB per year and the price is RM 1,

373,000 per year. Below is price for raw materials of this synthesis route;

17
Table 1. 7: Raw materials and their prices for production of 50 MTPA
PHB using soybean oil as carbon source.
Raw Material

Price

Price

Source

(RM/kg)

(RM/year)

Soybean oil

2.90

44, 900.00

MGT Group Bhd

Ammonium

10.35

12.40

Greymont Agrochem Sdn Bhd

sulphate

Economic potential level 1 of this synthesis route can be calculates as;

EP1:

RM 1, 373,000/year (RM 44, 900.00/year RM 12.40/year)

= RM 1, 330, 000.00 per year

1.4.1.2 Production of PHB from crude palm kernel oil (CPKO) by

Cupriavidus necator
Since the C. necator has the limitation on soybean oil as this cells grow
well on palmitic acid, linoeic and oleic acid but cannot grow well on linolenic
acid, the use of palm oil containing less linolenic acid may be a good choice to
accumulate a high dry cell weight and also could increase the yield of PHA
more than those with soybean oil (Kahar et al. 2008).
Today, Malaysia is both major producer and exporter of palm oil in the
world. Palm oil is a versatile oil that is currently used as edible oils as well as
for the production of oleo chemicals. Malaysia is one of the largest contributor
of palm oil in the world, surpassing Nigeria as the main producer since 1971
(Yusoff, 2006). Malaysia is the worlds second-leading oil palm producer and
exporter after Indonesia, supplying about 12.6% of global consumption of
vegetable oils (GAIN, 2012). This is firmly would support the supply of feed
stock for the PHA and PHB production.

18
However, there are issues requiring serious attention such as
deforestation, waste disposals from palm oil mill and energy expenditure when
PHA is to be produced in large scale. In order to fulfill the PHA market
demand solely by using CPKO to produce PHA, approximately 53,000 tons of
CPKO (which is approximately 2.8% of Malaysias total CPKO production) is
required as carbon feed stock for microbial fermentation. In other words, the
production of 52,000 tons of PHA per annum would involve a total of 111,520
hectares of oil palm plantation. As the demand for plant oils increase for PHA
production, it may result in the further expansion of plantations into forests.
Also, like soybean oil, there are concerns about merits of converting foodgrade oil for bio plastics production at the expense of dwindling the worlds
food supply as palm oil provides nearly 30% of the worlds edible vegetable
oil (Carter et al. 2007), with a production volume of 43.12 million tons in year
2008 (MPOB 2008).

1.4.1.2.1

Economic Potential Level 1

As we produce 50,000 kg of PHB per year and the price is RM 1,

373,000 per year. Below is price for raw materials of this synthesis route;
Table 1. 8: Raw materials and their prices for production of 50 MTPA
PHB using crude palm kernel oil (CPKO) as carbon source.
Raw Material

Palm kernel oil

Price

Price

(RM/kg)

(RM/year)

1.25

19, 350.00

Source

KL Kepong Oleomas Sdn

Bhd

Ammonia

8.00

10.20

Petronas

Chemicals

Ammonia Sdn.Bhd

Economic potential level 1 of this synthesis route can be calculates as;

EP1:

RM 1, 373, 000/year (RM19, 350.00/year RM 10.20/year)

19
= RM 1, 354, 600.00/year

1.4.1.3 Production of PHB from jatropha oil by Cupriavidus necator H16

As a result of evaluation of using edible plant oils, Jatropha oil, as a
non-edible one, would not affect the global food chain crisis and has potential
as a renewable resource. It can be the alternative substrate for bio plastic
production. Jatropha oil, derived from Jatropha curcas seed, also yield high
amounts of oil, yielding almost four times than the soybean and ten times from
the maize (Fitzgerald, 2006). It is also relatively cheap, costing less than
soybean oil as the fertilizer and pesticide requirement of Jatropha is lower
(Gui et al, 2008).
As the world is in a state of biofuels fever, many countries have started
planting Jatropha plant as this non-edible plant has promising future as
biofuels. Approximately 900,000 hectares of Jatropha have already been
planted throughout the world. Although the industry is in its early stages, it is
identified 242 Jatropha plantation projects, totaling approximately 900,000
hectares. More than 85% of the land cultivated is located in Asia. Africa
counts for approximately 120,000 hectares followed by Latin America with
approximately 20,000 hectares. Jatropha saw enormous growth: 5 million
hectares were expected by 2010. The number and size of Jatropha projects
currently being developed is increasing sharply. This is the case in almost all
regions of the world which are suitable for Jatropha cultivation. It is predicted
that each year for the next 5-7 years approximately 1.5 to 2 million hectares of
Jatropha will be planted. This will result in a total of approximately 5 million
hectares by 2010 and approximately 13 million hectares by 2015.

20

Figure 1. 5: Total acreage of jatropha oil plantations in selected countries

(Extracted from: http://www.jatropha-alliance.com)

Jatropha has been commercially farmed in Malaysia specifically

Sarawak owned by Bio Oil National Group Malaysia (The Star, 2011). A few
local private companies also have engaged in Jatropha cultivation scaling from
400 ha to 1000 ha. It has been identified that total current acreage of Jatropha
plantation projects is 1,712 ha. Project owners state plans to increase the
cultivation scale to a total of 57,601 ha by 2015. The Ministry of Plantation of
Industries and Commodities is undertaking a Jatropha pilot research project
for which 300 ha have been allocated.
The operational and maintenance costs for the Jatropha oil extraction
are very minimal, estimated at approximately 10 15% of the capital cost per
year. In Ghana, for instance, in 2010, whilst the cost of Jatropha oil and
kerosene were estimated to be US$0.085/liter and US$1.23/liter respectively,
the cost of

biodiesel from jatropha oil and petroleum diesel were also

estimated at US$0.99/liter and US$1.21/liter respectively (Ofori-Boateng and

21
Teong, 2011). On the other hand, the seeds from the northern part of Malaysia
contain high lipid content of 60% of oil (Salimon and Abdullah, 2008). Using
jatropha as carbon source in PHB production can reduce the recovery cost
(Choi and Lee, 1997) as it yields high PHB content.

1.4.1.3.1

Economic Potential Level 1

As we produce 50,000 kg of PHB per year and the price is RM 1,

373,000 per year. Below is price of raw materials of this synthesis route;
Table 1. 9: Raw materials and their prices for production of 50 MTPA
PHB using Jatropha oil as carbon source.
Price

Price

(RM/kg)

(RM/year)

Jatropha oil

2.73

19, 503.00

Bionas Malaysia Sdn Bhd

Urea

1.70

2.75

Petronas Fertilizers Kedah

Raw Materials

Source

Economic potential level 1 of this synthesis route can be calculates as;

EP1:

RM 1, 373, 000/year (RM 19, 503.00/year RM2.75/year)

= RM 1, 353, 000/year

22

Table 1. 10: Comparison between soybean oil, jatropha oil, and crude palm kernel oil based on its availability of raw materials, yield of
PHB, concerns, cost and operation mode.
Raw material

Jatropha Oil

Soy Bean Oil

Palm Oil

Microbial strain
Cupriavidus necator H16

used
Availability

of Malaysia

has

potential

for

jatropha There

is

raw materials in plantations because this plant grows more cultivation

Malaysia

no
of

commercial Malaysia is the second largest palm oil

soybeans

in producer and exporter in the world (Kek et

quickly and produces more seed in the Malaysia. Malaysia has to import al. 2011).
tropics (Openshaw, 2000).

soybean oil from United States

Jatropha seed from northen part of (GAIN, 2012).

Malaysia has a high lipid content of 60
wt% of oil (Salimon and Abdullah, 2008).
Yield of PHB

Yield high amounts of oil, yielding almost Yield almost two-fold higher than 68 wt% PHB content from crude palm
four times than the soybean and ten times from glucose (Akiyama et al., kernel oil (Kek et al. 2010)
from the maize (Fitzgerald, 2006)

Concerns

2003)

Jatropha oil is a non-edible oil would not The

use

of

edible

oils

in High demand for PHA production may

affect the global food chain crisis and has production of bio plastics may cause more deforestation for palm oil

23

potential as a renewable resource can be cause depletion of global food plantation.

the alternative substrate for bioplastic supply and sources.
production.
Cost

Jatropha plants are estimated to cost less Edible

plant

oils

price

has Palm oil can be obtained locally hence

than soybeans due to lower fertilizer and increased drastically because of reducing shipping and handling cost.
pesticides requirements (Gui et al. 2008)

recent crisis of food shortage and However, supply for PHA production has
increase of food demand. (Ng et to compete with demand of palm oil as
al. 2010)

Operation mode

Fed-batch fermentation

commercial edible oil.

24
1.4.2

Selected synthesis route

Based on evaluation from three synthesis routes, jatropha oil is chosen

as the main carbon source for PHB production with C. necator H16 as this
route provides more advantage in term of cost reduction and high yield of
PHB.

1.4.3

Utilization of Jatropha oil

The operational and maintenance costs for the Jatropha oil extraction

are very minimal, estimated at approximately 10 15% of the capital cost per
year. In Ghana, for instance, in 2010, whilst the cost of Jatropha oil and
kerosene were estimated to be US$0.085/liter and US$1.23/liter respectively,
the cost of

biodiesel from Jatropha oil and petroleum diesel were also

estimated at US$0.99/liter and US$1.21/liter respectively (Ofori-Boateng and

Teong, 2011). On the other hand, the seeds from the northern part of Malaysia
contain high lipid content of 60% of oil (Salimon and Abdullah, 2008). Using
Jatropha as carbon source in PHB production can reduce the recovery cost
(Choi and Lee) as it yields high PHB content.

1.4.4

Type of Microbial Production Strain

Several bacteria strains have been studied in accumulation of PHB.

Researches are conducted and microorganisms such as Alcaligenes latus

(Yamane et al., 1996), Alcaligenes eutrophus now is known as Cupriavidus
necator (Kim et al., 1994), Azotobacter vinelandii (Page and Knosp, 1989),
Pseudomonas oleovorans (Brandl et al. 1988), and recombinant Escherichia
coli (Lee and Chang, 1994). Lee et al., 1994 have showed some promising
high yield of PHB production. E. coli strains are considered as impractical in
large scale production of PHB as it is expensive. They require expensive
Luria-Bertani (LB) medium, ampicillin and pure O2 (Liu et al., 1998). E.coli is
also unable of producing PHAs, however it can utilise several carbon sources

25
including some substrates that cannot be easily used by most of the
microorganisms, such as lactose (Lee et al., 1997). Table 1.11 shows
comparison between promising microorganisms in PHB cultivation, an
analysis from Choi and Lee (1997). It is shown that PHB concentration, PHB
productivity and PHB yield are higher in C. necator.
Table 1. 11: comparison between promising microorganisms in PHB
cultivation, an analysis from Choi and Lee (1997).
Recombinant

Bacterium

C. necator

A. latus

Carbon source

Glucose

Sucrose

Glucose

Limiting nutrient

Nitrogen

None

None

Fermentation method

Glucose

pH-stat

pH-stat

E.coli

concentration
control
Culture time (h)

50

28.45

39

Cell concentration (g/L)

164

143

110

PHB concentration (g/L)

121

71.4

85

PHB content (%)

76

50

77.3

PHB productivity (g/L.h) 2.42

2.5

2.18

PHB yield (g PHB/ g 0.3

0.17

0.29

5.88

3.5

substrate)
kg substrate/kg PHB

3.33

Reference

Kim
1994

et

al. Yamane et al. Lee and Chang

1996

1994

Cupriviadus necator use up palmitic acids, oleic acids and linoleic

acids contained in jatropha oil. Freitas et al. (2009) reported that C. necator
has been proven to accumulate PHB up to 80% of its cell dry weight. Whilst
Khan et al. (2013) reported that cell growth curve of C. necator H16 has a
classical pattern with an exponential phase up to 50 hours followed by
stationary phase that lasted until 65 hours. They also stated that the highest

26
cell dry weight of 11.6 g/L was obtained at 55 hour followed by highest PHB
concentration of 8.6 g/L at 61.5 hour. Kadouri et al. (2005) reported that C.
necator can tolerate adverse stress conditions such as heat, osmotic pressure,
UV radiation and toxins such as ethanol and hydrogen peroxide. It should be
clear that C. necator is the most suitable microorganism to be used with
jatropha oil in PHB production.

1.4.5

Feeding source of nutrient supply

Lee et al. (2008) found that different nitrogen sources affected both

cell biomass and PHA biosynthesis. They also discovered that both urea and
sodium nitrate resulted in better biomass production compared to other
nutrient supply. Ng et al. (2010) stated that urea is the most suitable nitrogen
source to pair up with Jatropha oil as carbon source. Besides, urea costs much
lower price and has high productivity of PHB (Kek et al., 2008). Khanna and
Srivastava (2005) as well as Sabra and Abou Zeid (2008) discovered that urea
can yield high cell biomass and PHA production significantly. Ng et al. (2010)
also reported that based on their analysis both cell dry weight (CDW) and
PHB accumulation increased when the urea concentration increased. They also
stated that the optimal concentration of urea is 0.54 g/L as CDW remained
constant while PHB accumulation decreased significantly after 0.54 g/L of
urea.

1.4.6

PHB synthesis
There are four methods identified to cultivate PHAs which are; in

vitro, via PHA-polymerase catalyzed polymerization; and in vivo with batch,

fed-batch and continuous cultures (Zinn et al., 2001). However, the fed-batch
mode is the most used for PHAs production to achieve high cell density,
which often crucial for the high productivity and yield for the desired product.
Fed-batch mode is chosen as it has a lot of advantages in production of PHB.
Heuristically, fed-batch can maintain the carbon-source concentration at very

27
low concentration, to maximize the biomass yield. Ng et al. (2010) presented
Jatropha oil support the cell growth and PHB production in fed-batch
fermentation and high yield of product per Jatropha oil was obtained.
The fermentation process is relatively simple with multi-staging from
the petri dish to a shaker flask to a small fermenter which is then used to
inoculate the production reactor. In fermentation process, cells were
maintained and pre-cultivated in 2 g/L yeast extract, 10 g/L meat extract and
10g/L peptone (Khan et al., 2013).

1.4.7

Downstream Process
For subsequent process, several methods have been developed for the

recovery of PHAs, mostly PHB from the cells. Solvents such as chloroform,
methylene chloride, propylene carbonate and dichloroethane have been used
for the extraction of PHB (Ramsay et al., 1994). However, it was difficult to
remove the cell residues due to viscosity of 5% (w/v) PHB from extracted
polymer solution (Choi and Lee, 1997). Hahn et al. 1994 suggested that PHB
can be recovered using a dispersion of hypochlorite and surfactant solution.
PHB recovery by this method is more efficient with less polymer degradation.
During recovery of PHB, the harvested cell pellets are treated with
surfactant solution (1%, w/v) at 25C for 1 hour of mean residence time. The
treat mean is then followed by hypochlorite digestion in flow-through manner
and then PHB is separated from supernatant by centrifugation. PHB granules
are rinsed with water and were finally spray-dried. (Lee and Choi, 1997).

28

CHAPTER 2

PROCESS FLOW SHEETING

2.1

Selection of raw material and impurities management

In order to produce targeted amount of 50 metric ton per year of PHB,

and based on 8000 operation hours per year, the raw materials are decided
based on their high yield of product and economy wise, gives advantages to
cost of production. Based on its market availability, crude Jatropha oil is
chosen as the substrate. Urea is selected as nitrogen source as it could produce
high PHB content according to Khan et al. (2013). Meanwhile, the bacteria
strain that used is Cupriavidus necator H16 which previously known as
Ralstonia eutropha. It was obtained from National Collection of Industrial,
Food and Marine Bacteria.
Since we are using crude Jatropha oil with high purity, which also can
be obtained with affordable price locally, there is no need to manage impurity
of substrate. The oil, which contains high carbon source is used directly in
inoculation.

2.2

Input and Output Flow Sheeting

Input and output flow sheeting is illustrated in block flow diagram to

give clear view of the process. Figure 2.1 illustrates the flow of PHB
production while Figure 2.2 and Figure 2.3 shows upstream and downstream
process, respectively. Upstream process includes early cell isolation and
cultivation, to cell banking and culture expansion of the cells (fermentation)
until final harvest (termination of the culture and collection of the live cell
batch). Downstream process starts from harvesting from fermenter, and then
processed

to

meet

purity

and

quality

requirements.

30

Figure 2. 1: Block Flow Diagram of PHB Production

31

Figure 2. 2: Block Flow Diagram of Upstream Process

32

Figure 2. 3: Block Flow Diagram from Downstream Process

33

Figure 2. 4: Process Flow Diagram of PHB plant

34

Figure 2. 5: Process Flow Diagram Simulation in SuperPro

33
Figure 2.4 illustrates the process flow diagram based on block flow
diagram, fully labelled of streams and pumps. The upstream process started
with the sterilization of mineral medium and jatropha oil in ST-101 and ST102, respectively and mixed into mixing tank MX-101. The microbes
Cupriavidus necator H16 is inoculated in shake flask for 48 hours before
subjected to seed fermenter. Solution from mixing tank is splitted and 10%
will go to seed fermenter and the rest is fed into main fermenter. Air is
supplied using gas compressor (G-101) and filtered to avoid contamination in
fermenters using air filter AF-101. Vent from main fermenter is filtered using
air filter AF-102.
The downstream process utilizes the centrifugation process only. The
separation stage was based on methodology from Choi & Lee (1997) as well
as Choi & Lee (1999). After the end of the fermentation, the bacterial cells are
harvested via continuous centrifugation (DS-101) of the fermentation broth
which collected in V-103. Microbial cell lysis or disruption is carried out via
combined surfactant-hypochlorite digestion. A surfactant solution (Triton X100) of 1% (w/v) is added to the microbial biomass, charged in stream S-122
and mixed at 25C. This treatment followed by centrifugation in DS-102 to
separate PHB granules and aqueous solution and then added hypochlorite
digestion, charged in stream S-124 in a flow-through manner. This combined
step results in microbial lysis and separation of PHB from residual cell
material. The aqueous solution containing the residual cell material is
separated further by centrifugation in DS-103, where PHB is rinsed with water
beforehand V-105. Finally, PHB granules are purified via washing with water
and drying via a spray-drying step.

2.2.1

Mechanical Equipment Description

Based on preliminary flow diagram in Figure 2.1, there are thirteen

equipment used to produce high purity of 50 MTPA PHB. Follows are

description for each equipment used in this design.

34
2.2.1.1

Heat sterilizer (ST-101 & ST-102)

Two heat sterilizers are used to heat the raw materials media

continuously in 121C temperature before being sent to seed fermenter (V102) and main fermenter (V-103). Jatropha oil must be autoclaved separately
from mineral media. Sterilization is essential in bioprocess plant to avoid any
contamination during main process. These heat sterilizers have diameter of
0.10m and 7.42 m length.

2.2.1.2

Blending tank (V-101, V-105, & V-106)

Blending tank V-101 is installed to blend the sterilized Jatropha oil

from S-104 and sterilized mineral media from S-103. V-105 is used to blend
surfactant (S-122) with supernatant (S-121) while V-106 is used to blend
water from stream S-128 and PHB for washing and rinsing of granules. A
stainless steel 20 m3 blending tank with Siemens motors is used in this
process.

2.2.1.3

Disk Stack Centrifuge (DS-101, DS-102, & DS-103)

Centrifuge DS-101 is required to harvest the cells of fermentation

broth collected from storage by continuous centrifugation. DS-102 is used to

separate PHB from aqueous solution containing dissolved non-PHA cellular
material (NPCM) while DS-103 is used to rinse PHB granules with water.

2.2.1.4

Spray Dryer (SDR-101)

This last piece of equipment in this process is used to produce dry

powder from slurry consists of PHB granules and water (S-132). The product
is collected at S-133 while waste is channeled to waste treatment plant through
S-134. The design of this spray dryer is estimated with 0.80 m diameter with
2.39 m in height.

35
2.2.1.5

Fermenter (V-103)
Fermenter with fed-batch mode is chosen to produce PHB from

Jatropha oil by C. necator H16. Fermenter with volume 200 m3 is used in

order to produce 50,000 kg of PHB per year.

2.2.1.6

Gas Compressor (G-101)

Axial gas compressor is chosen to continuously pressurized air from

surrounding environment as this compressor has high efficiency and large

mass flow rate. The pressure change in this compressor is 2 bar.

2.2.1.7

Air filter (AF-101 & AF-102)

Air filter is one of the essential equipment in bioprocess plant. Air

filter AF-101 is connected after gas compressor function as to filter any

particulates coming from surrounding air. It is needed to filter to avoid any
contamination in the main fermenter that might alter the product. AF-102 is
installed to ensure no microorganism exit from fermenter as they might be
harmful and toxic to plant personnel and environment. The throughput is
estimated as 0.70 m3/s.

2.3

Material and Energy Balances

This section provides details of manual calculation of material and

energy balance, and simulation using Superpro software was conducted and
the results was compared accordingly with manual calculation.

36
2.3.1

Material Balance
In this section, we will perform material balance for each process unit

involved in this PHB production. Material balance is calculated by using

Microsoft Excel and the spreadsheet is as attached in Appendix A.1. The
overall material balance for the whole process is performed using the general
equation shown below:
Input + Generation Output Consumption = Accumulation
Input

total mass entering system boundaries

Generation

total mass produced within system

Output

total mass leaving system boundaries

Consumption

total mass consumed within the system

Accumulation

total mass built up within the system

Here, some assumptions have been made to make the calculation more
easily. The design-based assumptions are:
1. No leakage in the pipes and vessels in the system.
2. All the components in the system behave as ideal condition.

2.3.1.1

Heat Sterilizer (ST-101) & (ST-102)

No mass generation, consumption and accumulation occur within this

unit, therefore, we know that Input (S101) = Output (S104) and Input (S102) =
Output (S103)

Table 2. 1: Input and Output of Heat Sterilizer (ST-101&ST-102)

Material

Input = Output (kg/batch)

37
Jatropha Oils

389.828

Urea

0.235

Mineral Medium

153.76463

2.3.1.2 Blending Tank (V-101)

1. Jatropha oil
2. Mineral medium :
a. KH2PO4
b. Na2HPO4.12H2O
c. MgSO4.7H2O
d. Urea
e. Trace elements :
i. H3BO3,
ii. CoCl2.6H2O,
iii. ZnSO4.7H2O,
iv. MnCl2.4H2O,
v. Na2MoO4.2H2O,
vi. NiCl2.6H2O
vii. CuSO4.5H2O

38
Table 2. 2: Density of Each Components
Density (kg/m3)

Materials
Mineral Medium:

KH2PO4

2340 (Wikipedia, the free encyclopedia, 2014)

Na2HPO4.12H2O

1520 (LookChem.com, 2008)

MgSO4.7H2O

2660 (Fisher Scientific, 2000)

Urea

1320 (Wikipedia, the free encyclopedia, 2014)

Trace elements:

H3BO3,

1440 (Wikipedia, the free encyclopedia, 2014)

CoCl2.6H2O,

1920 (Fisher Scientific, 2001)

ZnSO4.7H2O,

1960 ( Fisher Scientific, 2001)

MnCl2.4H2O,

2010 (LookChem.com, 2008)

Na2MoO4.2H2O,

3780 (Wikipedia, the free encyclopedia, 2013)

NiCl2.6H2O

3550 (Acros Organics N.V., 2000)

CuSO4.5H2O

2280 ( Fisher Scientific, 2000)

Jatropha oil

920 (WeblinkIndia.NET, 2013)

Nutrient-rich medium:

Yeast

1100 (Andrea K. Bryana, 2010)

Meat extract

250 (Merck Chemical, 2014)

Peptone

250 (Merck Chemical, 2014)

In this project, we are going to produce 50 MTPA PHB,

50 MT

1 year

1 kg

year

8000hr

0.001 MT

= 6.25 kg PHB/hr

1 batch requires 61.5 hours,

6.25 kg

61.5 hr

Hr

batch

384.375 kg PHB/batch

39
Since the actual operating days is 333 days 8000 operating hours, therefore;
8000 hrs

1 batch

1 year

61.5 hrs

YP/S

= 98.7 %

130 batches/year

= Product / Substrate (Jatropha)

Substrate

384.375 kg

required =

Batch

0.987

389.438 kg Jatropha

/batch

*Assumption: Yield of feeds =Yield of substrate

From literature, (Khan, 2013)
1L of trace element solutions consists of

: H3BO3

= 0.0003kg/l

: CoCl2.6H2O = 0.0002kg/l
: ZnSO4.7H2O = 0.0001kg/l
: MnCl2.4H2O = 3x10-6kg/l
: Na2MoO4.2H2O = 3x10-6kg /l
: NiCl2.6H2O = 2x10-6kg /l
: CuSO4.5H2O = 1x10-6kg /l
H3BO3:
389.438kg
substrate
Batch

0.0003kg

1m3

1000L

1440kg

1m3

0.0002kg

1m3

1000L

1920kg

1m3

= 0.08113kg H3BO3/
batch

COCl2.6H2O:
389.438kg
substrate
Batch

= 0.04057kg CoCl2.6H2O
/ batch

40

ZnSO4.7H2O:
389.438kg
substrate

0.0001kg

1m3

1000L

1960kg

1m3

3x10-6 kg

1m3

1000L

2010kg

1m3

3x10-6 kg

1m3

1000L

Batch

= 0.01987ZnSO4.7H2O
kg/ batch

MnCl2.4H2O:
389.438kg
substrate
Batch

= 5.8125x10-4
MnCl2.4H2O kg/ batch

Na2MoO4.2H2O:
389.438kg
substrate
Batch

=3.0908x10-4
Na2MoO4.2H2O kg/

3780kg

1m

2x10-6 kg

1m3

1000L

3550kg

1m3

batch

NiCl2.6H2O:
389.438kg
substrate
Batch

= 2.1940x10-4
NiCl2.6H2O kg/ batch

41
CuSO4.5H2O:
389.438kg
substrate

1x10-6 kg

1m3

1000L

2880kg

1m3

Batch

= 3.2910x10-4
CuSO4.5H2O kg/ batch

Only 0.1% of trace elements are used (Khan, 2013). Therefore, total trace
elements used in blending tank is;
0.143kg trace elements/batch X 0.001 = 1.43 x 10-4kg trace elements /batch
1L of mineral medium consists of

: KH2PO4

= 1.5g/l

: Na2HPO4.12H2O

= 9g/l

: MgSO4.7H2O

= 0.2g/l

: Urea

= 1g/l

: Trace elements

= 1ml

KH2PO4:
389.438kg
substrate

0.0015kg

1m3

1000L

2340kg

1m3

Batch

= 0.2496kg KH2PO4
batch

Na2HPO4.12H2O:
389.438kg
substrate
Batch

0.009kg

1m3

1000L
=2.3059kgNa2HPO4.12H2O/batch

1520kg

1m3

42
MgSO4.7H2O:
389.438kg
substrate
Batch

0.0002kg

1m3

1000L

2660kg

1m3

0.001kg

1m3

1000L

=0.02928kg
MgSO4.7H2O /batch

Urea:
389.438kg
substrate
batch

=0.2950kg Urea/batch
L

1320kg

1m3

The total mineral medium required for this blending tank is 2.8799 kg/batch
Table 2. 3: Summary of materials used in blending tank

2.3.1.3

Materials

Amount of input (kg/batch)

Mineral medium

2.8799

Jatropha oil

389.438

Flow Splitter (FSP-102)

This unit splits the input stream (output stream from blending tank)

into 2 output streams. One stream is fed into seed fermenter (10%) while the
other stream goes into main fermenter (90%). Similarly, no mass generation,
consumption and accumulation occur within this unit, thus:
Input(S-105) = Output(S-106 & S-107)
Assumption: We assign 10% of the total amount into seed fermenter

43
Table 2. 4: Amount of input of Seed Fermenter and Main Fermenter
Amount of input (kg/batch)
Main Fermenter
Material

Total

Seed Fermenter

(V-103) (including

(V-102)

total output from

inoculum)

Jatropha oil

389.43

38.943

350.494

C.Necator(biomass)

1.597

1.1537x10-3

1.596

Mineral medium

0.028799

0.00288

0.0259

1.7272

1.727

6.906x10-4

Oxygen

2296.0288

229.6

2066.43

CO2

Water

PHB

39.457

39.457

Pre-culture medium
(include biomass)

2.3.1.4 Compressor (G-101)

Atmospheric air is compressed into the process. No mass generation,
consumption and accumulation occur within this unit, therefore, we know that:
Input (S108) = Output (S109)
Table 2. 5: Input and Output of Compressor (G-101)
Materials

Input = Output (kg/batch)

Air

22960.29

Oxygen

482.1930672

Nitrogen

1813.835933

44
2.3.1.5 Air Filter (AF-101)
We may assume that the filtered particles are of negligible weight.
Therefore, we consider that no mass generation, consumption and
accumulation within this unit.
Input (S109) = Output (S110)
Table 2. 6: Input and Output of Air Filter (AF-101)
Materials

Input = Output (kg/batch)

Air

22960.29

Oxygen

482.1930672

Nitrogen

1813.835933

2.3.1.6 Fermenter (V-102)

Material balance of fermenter is done for both the seed fermenter and
main fermenter. The overall balance for these two fermenters will be shown at
the end of this section.
From the journal (Khan, 2013), pre-culture medium consists of;
1. Jatropha oil

=1g/l

2. Yeast

=2g/l

3. Meat extract

=10g/l

4. Peptone

=10g/l

0.04ml of 1000ml= 0.004%

5. C. Necator

Mineral medium
Conditions:

= 10ml of 1000ml = 1%

45
Temperature of culture medium

: 37oC

Fermentation duration, t

: 61.5 hours

Yp/s

: 98.7% *

Yx/s

: 0.0026 g/g*

*From journal (Khan, 2013) 98.7% complete conversion of substrate to

product
In this section, stoichiometric parameter, yield is used to describe
growth kinetics. Yield coefficients are defined based on the amount of
consumption of another material. Yx/s (g cell/g substrate) is defined as amount
of cell to amount of substrate required. Yp/s is defined as the ratio of product
produced to substrate consumed.

2.3.1.7 Seed Fermenter (V-102)

From literature (Ko-Sin Ng, 2010), 1g/l of Jatropha oil were added to
the seed fermenter together with C. Necator, yeast, meat extract and peptone.
It is about 10% from overall substrate used. Jatropha oil was added to the preculture to induce lipase secretion for better oil utilization for cell growth and
higher P (3HB) production.
389.438 kg substrate

10%

batch

100%

= 38.9438 kg Jatropha oil/ batch

Now, we proceed to the material balances involving production of

PHB from C. Necator. First we need to calculate the value of PHB and
Jatropha in term of mol/batch.
384.375 kg PHB

kmol

1000mol

= 4469.48 mol PHB

batch

86 kg

1 kmol

/batch

46

4469.48mol PHB

1 mol Jatropha oil

batch

3.23 mol of PHB

Jatropha /batch

1383.74mol

From literature, (Khan, 2013) Yx/s = 0.0026g cell/g substrate, thus total
amount of C.Necator leaving the seed fermenter is:
0.0026 kg cell

282 kg

kmol

= 0.0298 mol biomass/

kg substrate

kmol

24.6 kg

batch

For 50 MT,
0.0298 mol

1383.74mol

biomass

Jatropha oil

batch

kg
25.83mol

= 1.596kg biomass/
batch

For every C. Necator used,

Yeast:
1.596 kg cell

0.002kg

1m3

1000L

= 2.902x10-3kg yeast

batch

1100kg

1m3

/batch

Meat extract:
1.596 kg cell

0.01kg

1m3

1000L

= 0.0638kg meat extract

batch

250kg

1m3

/batch

47
Peptone:
1.596 kg cell

0.01kg

1m3

1000L

batch

250kg

1m3

/batch

0.0638kg

peptone

We feed in 10% of the substrate into seed fermenter and we predict the
amount of PHB produced in cell in seed fermenter as:
389.438 kg substrate

10

Batch

100

0.987

39.457kg PHB/batch

*1% of mineral medium used in fermenter

Therefore, total mineral medium used in fermenter:

= 2.8799kg/batch X 0.01
= 0.028799 kg mineral medium/batch

Since only 10% of the mineral medium used in seed fermenter, thus the
amount of mineral medium input at the seed fermenter is 0.028799 x 0.1 =
0.00288kg mineral medium/ batch

Table 2. 7: Summary of material used in seed fermenter

Materials

Input (kg/batch)

C. necator

1.597

Yeast

2.902x10-3

Meat extract

0.0638

Peptone

0.0638

Jatropha oil

38.943

Mineral medium

0.00288

48
2.3.1.8 Main Fermenter (V-103)
From literature (Khan, 2013),

0.04% of pre-culture is used into the fermenter.

Therefore, total pre-culture inoculated into fermenter:
=( 1.596kg biomass/ batch + 2.902x10-3 kg yeast /batch +0.0638kg meat
extract /batch + 0.0638kg peptone /batch) X (0.04/100)%
=6.906x10-4 kg pre-culture medium/batch

1% of mineral medium used in fermenter

Therefore, total mineral medium used in fermenter:
=2.8799kg/batch X 0.01
=0.028799 kg mineral medium/batch

Since only 90% of the mineral medium left, thus the amount of mineral
medium input at the fermenter is 0.028799 x 0.9 = 0.02592kg mineral
medium/ batch

Only 90% of the Jatropha oil used in fermenter:

=389.438 kg Jatropha oil / batch x 0.9 = 350.494kg Jatropha oil/ batch

Table 2. 8: Summary of materials used in main fermenter

Material

Input(kg/batch)

Pre-culture medium

6.0746x10-4

Mineral Medium

0.028799

Jatropha oil

350.49

We consider the following stoichiometric equations to calculate the

amount of oxygen, water and carbon dioxide involved in this process.
(C4H6O2: monomer of PHB)

49
Assumption: Negligible production of PHB during fermentation and no other
side reactions occur during fermentation. Molecular weight of PHB is
282kg/kmol.

Stoichiometric Equation:
According from journal (Khan, 2013):
The Cupriavidus necator strain was used to synthesize PHB by using Jatropha
oil as carbon source.
C18H34O2 + a O2 + bCH4N2O

c CH1.8O0.5N0.2 + d CO2 + e H2O +

Bacteria strains:

Cupriavidus necator

Carbon source:

Jatropha oil

Nitrogen source:

Urea

C4H6O2

Molecular weight of biomass:

CH1.8O0.5N0.2

= 12 + (1)(1.8) + 16(0.5) + 14(0.2)

= 24.6 g/mol *Assume that it contain ash about 5%.

Molecular weight of biomass: 24.6 + (0.05)(24.6)

= 25.83 g/mol

Molecular weight of substrate:

C18H34O2 = 12(18) + 1(34) + 16(2)

= 282 g/ mol

Molecular weight of product:

C4H6O2 = 12(4) + 1(6) + 16(2)

= 86 g/ mol

50
Degree of Freedom
6 number of unknown components/stoichiometric coefficients (a,b,c,d,e,f)
-4 number of elements balance (C, H, O, N)
-2 additional equations (YP/S, electron balance equation)
0 DOF
Elemental Balances:
C: 18 + b = c + d + 4

--------- (1)

H: 34 + 4b = 1.8c + 2e +

--------- (2)

O: 2 + 2 a + b = 0.5c + 2d + e +2

--------- (3)

N: 2b = 0.2c

--------- (4)

According to (Khan, 2013)

Yield of product over substrate consumed:

f = 3.2364

Yield of biomass over substrate consumed:

--------- (5)

51

c = 0.0288

---------- (6)

For element N:
2b

= 2c

= 0.0028

For element C:
18 + b

= c + d +4f

18 + 0.1c c - 4f

=d

18 + (0.1-1)0.028 4(3.23)

=d

d = 5.0548

--------- (7)

For element H:
34 + 4b = 1.8c +2e +6f
34 + 4(0.0028) = 0.0504 + 2e + 19.38
e = 7.2904

--------- (8)

For element O:
2 + 2a + b = 0.5c +2d +e +2f
2 + 2a + 0.0028 = 0.5(0.028) + 2(5.0548) + 7.2904

52
a = 7.7056

--------- (10)

a = 7.7056
b = 0.0028
c = 0.028
d = 5.0548
e = 7.2904
f = 3.234

The overall balance equation is:

C18H34O2 + 7.7056O2 + 0.0028CH4N2O

0.028CH1.8O0.5N0.2 +

5.0548CO2 + 7.29047H2O + 3.2364C4H6O2

From the stoichiometry above,
1 mole of

(Jatropha oil) equals to 1382.367 mol of Jatropha oil, so

the mole balance equation:

For 50 MT,

Ratio of O2 to N2 in air is 21% by volume O2 and 79% by volume N2.

Therefore, N2/O2 = 3.7619. From literature, we know that the residence time
of seed culture in seed fermenter is 20 hours. Besides, inoculum consists of
10% total volume of culture medium in the main fermenter, so, total air
required for seed fermenter is also 10% of the air required by main fermenter:

53
(1+3.7619)

15.0677 kmol O2

10%

1 batch

100%

7.1751 kmol oxygen

Meanwhile, total air required for main fermenter is:

(1+3.7619)

15.0677 kmol O2

90%

1 batch

100%

64.5758 kmol oxygen/hr

Hence, total amount of air = 71.7509 kmol x 32kg/kmol

= 2296.0288 kg O2
Since this is an aerobic reaction inside fermenter, we will supply more air than
the calculated amount of air.
*Assumption: Amount of air is 10 times the calculated values.
Therefore, amount of air input = 20807.761 kg air/hr
CO2 and H2O are produced during fermentation. For every batch:
CO2 produced

= 6.9509 kmol CO2/batch

= 6.9509 kmol CO2/batch x 44kg/kmol = 305.8396 kg

CO2/batch
H2O produced

= 10.0345 kmol H2O/batch

= 10.0345 kmol H2O/batch x 18 kg/kmol
= 180.6209 kg H2O/batch

Therefore, total H2O leaving fermenter = 180.6209 kg H2O/batch

Hence, total amount of air = 1806.209 kg air/cycle
CO2 and H2O are produced during fermentation.

54
For every batch (from mole balance calculation):
CO2 produced = 305.8396 kg/batch
H2O produced = 180.6209 kg/batch
Therefore, total H2O leaving fermenter = 180.6209 kg/batch

To sum up, we may summarize all the feeds into seed fermenters and main
fermenters:
Table 2. 9: Summary amount of input into Seed Fermenter and Main
Fermenter
Amount of input (kg/batch)
Main
Fermenter

Material
Total

Seed Fermenter

(V-102)

(V-101)

(including total
output from
inoculum)

Jatropha oil

389.43

38.943

350.494
-3

C.Necator(biomass)

1.597

1.1537x10

1.596

Mineral medium

0.028799

0.00288

0.0259

1.7272

1.727

6.906x10-4

Oxygen

2296.0288

229.6

2066.43

CO2

Water

PHB

39.457

39.457

Pre-culture medium
(include biomass)

Similarly, we summarize all the outputs leaving fermenters:

*Assumption: All nutrients are consumed completely by C.Necator

55
Table 2. 10: Summary amount of output from fermenter
Amount of output (kg/batch)

Materials

Seed Fermenter

Main fermenter

Jatropha oil

1.947

17.525

Water

18.0621

162.559

Mineral medium

Pre-culture (include biomass)

CO2

30.584

275.256

C. Necator

1.596

1030.7

Oxygen

PHB

39.457

384.375

To conclude, overall material balance of seed fermenter (V-102) is shown

below:
Table 2. 11: Overall material balance of Seed Fermenter
Amount (kg/batch)
Input

Input

Input

Output

S-106

S-111

S-114

S-115

Na2HPO4.12H2O

0.23059

KH2PO4

0.02496

MgSO.7H2O

0.002928

CH4N2O (Urea)

0.0295

Trace elements

0.0000143

Jatropha oil

38.9438

1.947

PHB

39.457

Oxygen

229.6

Water

18.0621

CO2

30.584

C.Necator (Biomass)

unknown

1.596

1.727

Materials

Pre-culture medium
(including biomass)

56

As for overall material balance of main fermenter (V-103) is shown below:

Table 2. 12: Summary of Overall Material Balance of main fermenter

Amount (kg/batch)
Materials
Na2HPO4.12H2
O
KH2PO4
MgSO.7H2O
CH4N2O (Urea)
Trace elements

Input S-

Input S-

Input S-

Output S-

Input S-

107

112

115

116

118

0.0207531

0.0207531

0.0022464

0.0022464

0.0002635
2
0.002655
0.0000012
87

0.0002635
2
0.002655
0.0000012
87

Jatropha oil

350.494

1.947

17.525

PHB

39.457

384.76

Oxygen

2066.43

Water

18.0621

180.6341

CO2

30.584

30.584

Nitrogen

16324.77

16324.77

1.596

1.013552

C.Necator
(Biomass)
Pre-culture
medium

2.3.1.9 Air Filter (AF-102)

We may assume that the filtered particles are of negligible weight.
Therefore, we consider that no mass generation, consumption and
accumulation within this unit.
Input (S116) = Output (S117)

57
Table 2. 13: Input and output of Air Filter (AF-102)
Materials

Input = Output (kg/batch)

Air (23wt% O2; 77wt% N2)

2296.0288 kg O2

2.3.1.10 Flat Bottom Tank (V-104)

Flat bottom tank are used for holding the broth liquids from main
fermenter. No mass generation, consumption and accumulation occur within
this unit, thus:
Input(S-118) = Output(S-119)
Table 2. 14: Input and Output of Flat Bottom Tank (V-104)
Flat Bottom Tank (V-104)

Material

Input

Output

PHB

384.375

384.375

Biomass

1.01355

1.01355

Nutrient Medium

392.7074

392.7074

Water

180.6341

180.6341

Total (kg/cycle)

958.7301

958.7301

2.3.1.11 Disk-Stack Centrifuge (DS-101)

Output stream from flat bottom tank is fed into downstream processing
unit centrifugation. Separation of medium (water + biomass) and cell with
PHB will be carried out in this unit operation. The mass balance for
centrifugation is summarized as follows:
*Assumptions:
1. Well-mixed and constant holdup in filter.
2. Efficiency = 0.90

58
Table 2. 15: Input and output of Centrifugal (C-101)
Centrifugal (C-01)

Material
Input(S-119)

Output(S-121) (90%

Output(S-120)

left)

(10% removed)

PHB

384.375

345.9375

38.4375

Biomass

1.01355

0.912195

0.10135

392.7074

353.4367

39.2707

180.6341

162.5707

18.0634

958.73

862.857

95.873

Nutrient
Medium
Water
Total
(kg/cycle)

2.3.1.12

Blending Tank (V-105)

For recovery step, sequential surfactant-hypochlorite will be used instead

of chloroform-hypochlorite method. The advantages are:

High quality of PHA

Rapid recovery and simple process

Retain native order of polymer chains in PHA granules

Lower operating cost compared to solvent extraction

Ramsay et. al. (1990) stated when PHB that containing biomass was
treated at a ratio of 1:1 with surfactant Triton X-100 treatment was followed
by a hypochlorite wash, a higher purity of 98% was achieved. Hence,
Amount of cell with PHB

= 345.9375/batch

Amount of surfactant used

= (2 345.9375) kg/batch
= 691.875 kg/batch

Purification yield: 98%

59
Table 2. 16: Input and Output of Blending Tank (C-101)
Blending Tank (C-101)
Material

PHB in triton
X
Biomass
Nutrient
Medium
Water
Total
(kg/cycle)

2.3.1.13

Output (S-123) (98 %

Output (S-124) (2%

left)

removed)

691.875

678.0375

13.8375

0.912195

0.89395

0.0182439

353.4367

346.3679

7.068734

162.5707

159.3193

3.251414

1208.7946

1184.6187

24.1759

Input(S-121)

Mixer (P-12/MX-101)

There is no mass generation, consumption and accumulation occurs

within this unit. Therefore,
Input(S-123) = Output(S-125)
From literature, we found that the hypochlorite is used to disrupt cell
wall of Cupriavidus necator in order to extract PHB from cell. Thus, we mix
an organic solvent (hypochlorite) in a ratio of 1 portions of hypochlorite to 1
portion of cell with PHB. Hypochlorite is mixed with cell in retentate stream
from MF1 (Jong, 1997). Hence,
Amount of cell with PHB in hypochlorite

= 678.0375kg/batch

Amount of hypochlorite used

= (2 678.0375) kg/batch
= 1356.075 kg/batch

Input and output streams of mixer MX-101 are summarized as follows:

Table 2. 17: Input and output streams of mixer (MX-101)

60

Material

Mixer (M-101)
Input(S-123)

Input (S-124)

Output (S-125)

PHB

678.0375

1691.6344

Biomass

0.89395

0.89395

Nutrient Medium

346.3679

Water

159.3193

159.3193

Hypochlorite

1356.075

Cell Debris

688.8461

Total (kg/cycle)

1184.6187

1356.075

2540.6933

Once hypochlorite is mixed with cell, it will disrupt the cell wall and
release PHB from cell. Then PHB dissolves in hypochlorite (solubility of PHB
in hypochlorite = 97%) (Sei, 1993).Referring to material balances in MF1
(345.9375PHB/batch), we know that the total amount of PHB dissolved in
hypochlorite is 335.5594 kg PHB/batch. In brief, there is one output stream
from mixer and it contains PHB in hypochlorite and water as well as cell
debris.

2.3.1.14

Disk-Stack Centrifuge (P-13/DS-102)

Output stream from flat bottom tank is fed into downstream processing
unit centrifugation. Separation of medium (cell debris) and cell with PHB
will be carried out in this unit operation. The mass balance for centrifugation
is summarized as follows:
*Assumptions:
1. Well-mixed and constant holdup in filter.
2. Efficiency = 0.90

61

Table 2. 18: Input and Output Stream of Disc-stack Centrifuge (P-13/DS102)

Material

Disc-stack Centrifuge (P-13/DS-102)

Input(S-125)

Output (S-127)

Output(S-126)

PHB in hypochlorite

1691.6344

1522.47096

169.16344

Biomass

0.89395

0.80456

0.089395

Nutrient Medium

Water

159.3193

143.3874

15.93193

Cell Debris

688.8461

68.88461

619.9615

Total (kg/cycle)

2540.6933

1735.54753

805.1463

2.3.1.15

Blending Tank (P-14/V-103)

For washing step, water will be used instead of chloroform-hypochlorite

method. The advantages are:

High quality of PHA

Rapid recovery and simple process

Retain native order of polymer chains in PHA granules

Lower operating cost compared to solvent extraction

Ramsay et. al. (1990) stated when PHB that containing biomass was
washed at a ratio of 1:5 with water, a higher purity of 98% was achieved.
Hence,

Amount of cell with PHB

= 1831.0299kg/batch

Amount of water used

= (5 1831.0299) kg/batch
= 9155.1495 kg/batch

Assume Purification yield: 98%

62

Table 2. 19: Input and Output Stream of Blending Tank (P-14/V-103)

Blending Tank (P-14/V-103)
Output
Material

Input

Input

Output

(remain in

(S-127)

(S-128)

(S-129)

blending
tank)

PHB in

1831.0299

1794.4093

36.6206

Biomass

0.9873

0.967554

0.019746

Water

175.9681

9155.1495

9209.8131

186.6224

Cell Debris

9.9213

9.7923

0.198426

2761.24444

9155.1495

hypochlorite

Total
(kg/batch)

2.3.1.16

11678.0660
6

238.3279

Disk-Stack Centrifuge (P-15/DS-103)

Output stream from blending tank is fed into downstream processing

unit centrifugation. Separation of medium (water) and cell with PHB will be
carried out in this unit operation. The mass balance for centrifugation is
summarized as follows:
*Assumptions:
1. Well-mixed and constant holdup in filter.
2. Efficiency = 0.90
Table 2. 20: Input and output of Disc-stack Centrifugal (C-03)
Material
PHB in hypochlorite

Disc-stack Centrifugal (P-15/DS-103)

Input(S-129)

Output (S-131)

Output (S-130)

1794.4093

1614.9684

179.4409

63
Biomass

0.967554

0.8708

0.106754

Water

9209.8131

920.98131

8288.83179

cell debris

9.7923

0.97923

8.81307

11678.06606

2537.79974

9140.26632

Total
(kg/batch)

2.3.1.17

Spray Dryer (P-16/SDR-101)

Permeate stream from S-131 is fed into spray dryer. Hot air(S-132) is
used to dry PHB. The desired product (PHB) will be in powder form.
Hypochlorite and water will be condensed and then are recycled. The input
streams of SDR-101 consist of:
Table 2. 21: Summary Input Stream of Spray Dryer (P-16/SDR-101)
Input S-132

Materials

Input S-131 (kg/batch)

PHB

316.6493

hypochlorite

1298.3191

Biomass

0.8708

Water

920.98131

Cell debris

0.97923

Air (23wt% O2; 77wt% N2)

(kg/batch)

4018.13100

Yield in this unit operation is 98.7%. This is because some of the PHB
powder may retain in dryer wall. Therefore, the final amount of PHB (in
powder form) produced is (0.987 x 316.6493 kg/batch) = 312.5329 kg/batch =
5.0818 kg PHB/hr. We summarize the output stream in the following tables:
*Assumption: All the water and hypochlorite are recovered.

Table 2. 22: Summary Output of Spray Dryer (P-16/SDR-101)

64
Output S134

Output S133

(kg/batch)

(kg/batch)

PHB (dry powder)

312.5329

Hypochlorite

1298.3191

Water

920.98131

Air

4018.131

Materials

2.3.2

Energy Balance
The energy balance calculated in this particular proposal will be

performed in the form of heat duty of the equipment. However, the heat duty
of certain equipment is equal to zero. This is because there is neither heat lost
nor heat generated throughout the process. For example, flow splitter and air
filter. We are going to perform energy balances for each stream and each unit
operation. While summary of heat duty of equipment will be shown later.
Basically, energy balance is calculated based on ideal condition. Other
assumptions include:

Heat generated by ions due to temperature difference is negligible.

There is no heat lost in the pipelines.

Potential and kinetic energy are neglected. We consider only the

enthalpy change.

Datum: 298.15 K and 1 atm

Steady state condition in all equipment.

Standard heat of formation of Jatropha oils is assumed equal to heat of

formation of vegetable oil since we are not able to get their heat of
solution.

Heat of formation of air is negligible. Heat of formation of PHB is set

equal to heat of combustion of bacteria.

For PHB in hypochlorite (solution), heat capacity of PHB equals to

heat capacity of hypochlorite.

65
We calculate the energy balance using spreadsheet which is attached in
Appendix A.2. Summary of energy balance for each stream is shown below
(please refer to Appendix A.2 for spreadsheet):
Table 2. 23: Summary of energy balance of each stream
Stream

Temperature (K)

Enthalpy (J)

101

298.15

-1306639.85

102

298.15

-1180029725.77

103

303.15

-1176326364.36

104

303.15

3548.75

105

303.15

-1172440460.13

106

303.15

0.00

107

303.15

0.00

108

298.15

-7174296.70

109

313.15

-7174296.70

110

310.15

0.00

111

298.15

0.00

112

303.15

0.00

113

298.15

0.00

114

298.15

0.00

115

303.15

-292247111.41

116

303.15

-7381773182.23

117

303.15

0.00

118

303.15

129087110064.21

119

303.15

127408976385.49

120

303.15

1678133726.32

121

303.15

127408973282.93

122

298.15

0.00

123

303.15

127408897039.05

124

303.15

-759539.99

125

303.15

21089163.69

126

298.15

991627914.63

127

303.15

3616247.11

66
128

298.15

5735499.15

129

303.15

83140088.79

130

303.15

6956135225

131

303.15

-13573749644.68

132

303.15

-36773140717.38

133

388.15

-351601803.28

134

303.15

130313154.62

Now, we proceed to the calculation for each equipment. Spreadsheet is

attached in Appendix A.2.
Gas Compressor G-101
Total heat duty for gas compressor is 0 kJ. All the steps of calculation
are shown in Appendix A.2.

Heat Sterilization ST-101

The total heat duty of sterilizer is 1310.188603 kJ. All the manual
calculations are carried out by using spreadsheet. Spreadsheet is attached in
Appendix A.2.

Heat Sterilization ST-102

The total heat duty of sterilizer is 3703.361416 kJ. All the manual
calculations are carried out by using spreadsheet. Spreadsheet is attached in
Appendix A.2.

67
Air Filtration AF-101
The total heat duty of sterilizer is 7174.296704 kJ. All the manual
calculations are carried out by using spreadsheet. Spreadsheet is attached in
Appendix A.2.

Air Filtration AF-102

The total heat duty of sterilizer is 7381773.182 kJ. All the manual
calculations are carried out by using spreadsheet. Spreadsheet is attached in
Appendix A.2.

Seed Fermenter V-101 and Main Fermenter V-102

According to Elementary Principles of Chemical Processes (by
Richard M. Felder and Ronald W. Rousseau), energy balance involved in
fermenter is expressed as:
Q Ws = H
where Ws refers to the impeller shaft work and H is enthalpy change. H can
be calculated using two methods: heat of reaction method and heat of
formation method. As we know that for heat of reaction method, the
expression is:
H = H r0 mout H out min H in
While for heat of formation method, the expression is:
H = mout H out min H in
In our study, we are not able to get the data of heat of reaction of the
process occurring in fermenter because we have no stoichiometry data on the

68
fermentation. Therefore, we use the heat of formation method and the energy
balance around the fermenter becomes:
Q = Ws + mout H out min H in
Table 2. 24: Heat of formation
Heat of formation

Material

MW (g/mol)

Na2HPO4.12H2O

1721.52

21798.6

KH2PO4

136.086

19664.8

MgSO.7H2O

120.336

-1374778.7

CH4N2O (Urea)

60.06

-333800

Jatropha oil

282

-853629.792

PHB

6600000

-17795728.250

Water

18.00

-285830.000

CO2

44.00

-393510

Biomass

25.83

466.7

Nitrogen

28.00

-12191.96

(J/mol)

Seed Fermenter V-101

First of all, we calculate the energy balance in seed fermenter. The
steps of calculation are shown in a spreadsheet attached in Appendix A.2.
Here, the heat duty of seed fermenter = -3164170.573 kW (total power need to
be removed from fermenter).

Main fermenter V-102

Similarly, we calculate the energy balance in main fermenter using the
same method. The steps of calculation are shown in a spreadsheet attached in
Appendix A.2. Here, heat duty of main fermenter = -5.40817 E +12 kW (total
power need to be removed from fermenter).

69

Blending Tank V-104

Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2.. Here, heat duty across microfilter = -1678133.679
kJ.

Blending Tank V-105

Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across mixer = -4770.795927 kJ.

Blending Tank V-106

Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across mixer = -4770.795927 kJ.

Centrifuge DS-101
Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across microfilter = 1678130.624
kJ.

Centrifuge DS-101
Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across microfilter = 1678130.624
kJ.

70
Centrifuge DS-102
Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across microfilter = 987616.301kJ.

Centrifuge DS-103
Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty across microfilter = 13587328.43122kJ.

Spray Dryer SDR-101

Heat duty of this equipment is calculated using spreadsheet which is
attached in Appendix A.2. Here, heat duty for dryer = 50125601.76340 kJ.
To conclude, heat duty of each equipment is summarized in Table 2.7.

Table 2. 25: Heat duty for each equipment

Components

Heat of duty (kJ)

Heat sterilizer ST-101

1310.188603

Heat sterilizer ST-102

3703.361416

Compressor G-101

0.00000

Air filtration AF-101

7174.296704

Air filtration AF-102

7381773.182

Blending Tank V-101

3882.355479

Seed Fermenter V-102

-3164170.573

Main fermenter V-103

-5.40817E+12

Blending Tank V-104

-1678133.679

Blending Tank V-105

-76.24388437

Blending Tank V-106

184751.8426

71

2.4

Centrifuge DS-101

1678130.624

Centrifuge DS-102

987616.301

Centrifuge DS-103

-13587328.43122

Spray Dryer SDR-101

50125601.76340

Economic Potential
Economic potential 1 has been calculated in Chapter 1 to show which

synthesis route is more feasible for this 50 MTPA PHB production. Economic
potential 2 and economic potential 3 are calculated further later.

2.4.1

Economic Potential 2 Based On Input and Output Structure

Figure 2. 6: Input-output structure of PHB production process

Figure 2.4 shows input and output structure of PHB process plant
based on 8000 operations hours per year. Conversion of Jatropha oil, XJ, the
reaction rate of PHB and molar fraction of CO2 at the vent are the design
variables. Data report from the empirical are the initial glucose concentration,
SJo, the concentration of urea, SUo, and the initial concentration of ethanol, SEo.
Mass balance is calculated using this following equations.

72
Jatropha oil balance,
(

FCJ +

(2.1)

(2.2)
(2.3)
(2.4)
(2.5)
(2.6)
(2.7)

Substitute Eq. (2.4) and Eq. (2.7) into Eq. (2.2)

F=[

)]

(2.8)

(2.9)

By using all Equation 2.1 until Equation 2.9, data is formulated as

shown in Appendix A.2. Then graph is plotted as below. Graph shows the
concentration of P (Polyhydroxybutyrate), X (biomass), J (Jatropha oil), and U
(urea) versus conversion of Jatropha oil at certain period.

73

Jatropha oil conversion

Concentration

0.2

0.4

0.6

0.8

20.0
19.5
19.0
18.5
18.0
17.5
17.0
16.5
16.0
15.5
15.0
14.5
14.0
13.5
13.0
12.5
12.0
11.5
11.0
10.5
10.0
9.5
9.0
8.5
8.0
7.5
7.0
6.5
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0

P (g/L)
X(g/L)
J(g/L)
U(g/L)

Figure 2. 7: Graph of concentration versus conversion of Jatropha oil

The economic potential level 2 can be defined as;

= Revenue F (Raw Material Cost)
= P1M1- F (CJoMJ+CUoMU)
Where M = price of raw materials MYR/kg, CJo = initial concentration
of Jatropha oil, CUo = initial concentration of urea, F = production rate per year
(L/year), EP2 = economic potential level 2 (MYR/year), P1 = PHB production
(kg/year). Calculation of EP2 is tabulated in Table 2.26 and graph for EP2 as
in Figure 2.1 is plotted to show the relationship of EP2 and conversion of

74
Jatropha oil, XJ. The graph shows that the project is feasible starting from
conversion at approximately 62%.
Table 2. 26: Values for EP2 calculation
t

XJ

P (g/L)

X(g/L)

J(g/L)

U(g/L)

EP2 (MYR/year)

0.000

3.300

0.053

20.000

1.000

1372500.000

10

0.111

3.667

0.058

17.778

0.889

-6304772.727

20

0.222

4.033

0.064

15.556

0.778

-2466136.364

30

0.333

4.400

0.070

13.333

0.667

-1186590.909

40

0.444

4.767

0.076

11.111

0.556

-546818.182

50

0.556

5.133

0.082

8.889

0.444

-162954.545

60

0.667

5.500

0.088

6.667

0.333

92954.545

70

0.778

5.867

0.093

4.444

0.222

275746.753

80

0.889

6.233

0.099

2.222

0.111

412840.909

90

1.000

6.600

0.105

0.000

0.000

519469.658

600000.00
500000.00

EP2 (MYR/year)

400000.00
300000.00
200000.00
100000.00
0.00
0
-100000.00

0.2

0.4

0.6

0.8

Jatropha conversion, XJ

Table 2. 27: Graph of EP2 versus Jatropha oil conversion

75
2.4.2

Economic Potential 3 Based On Recycle Structure

Figure 2. 8: Diagram of Recycle

Figure 2.8 shows a block flow diagram that represents PHB production
process with recycle streams. Culture fluid from chemo stat is channelled into
separator. The separation process will produce biomass in another stream and
cell-free supernatant in another stream. Biomass is then recycled back into
chemo stat. Figure 2.9 shows a graph to show the relationship of media and
biomass concentration with addition of recycled biomass concentration versus
conversion of Jatropha oil in certain time. It is clear that recycled biomass
(XR) has slightly larger concentration that X.

76

Jatropha oil conversion

Concentration

0.2

0.4

0.6

20.0
19.5
19.0
18.5
18.0
17.5
17.0
16.5
16.0
15.5
15.0
14.5
14.0
13.5
13.0
12.5
12.0
11.5
11.0
10.5
10.0
9.5
9.0
8.5
8.0
7.5
7.0
6.5
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0

0.8

P (g/L)
X(g/L)
J(g/L)
U(g/L)
XR g/L

Figure 2. 9: Graph of product, biomass, recycled biomass, Jatropha oil,

and urea concentration versus Jatropha oil conversion.

The economic potential level 3 can be defined as;

= Revenue F(Raw Material Cost) Reactor
cost
= P1M1- F (CJoMJ+CUoMU) Reactor Cost

77
The data of calculation of economic potentials at the second level (EP2),
at the third level with recycle (EP3+recycle), and at the third level without
recycle (EP3-recycle) are tabulated in Table 2.27. Further calculation is shown
in Appendix A.3. The relationship between EP2, EP3+recyle, and EP3-recycle
with conversion of Jatropha oil is shown as a graph in Figure 2.8.
Considerably, the process using the recycle makes more profit.

Table 2.27: Table 2. 28: Data for EP2 and EP3 at both with recycle and
without recycle.
t

XJ

EP2 (MYR/year)

EP3+recycle

EP3-recycle

10

0.111

-6304772.727

-2529819.1

-2582962.8

20

0.222

-2466136.364

-599176.64

-635714.22

30

0.333

-1186590.909

47712.257

18359.411

40

0.444

-546818.182

372437.47

347305.49

50

0.556

-162954.545

567921.82

545639.79

60

0.667

92954.545

698626.56

678430.64

70

0.778

275746.753

792233.78

773647.97

80

0.889

412840.909

862609.35

845313.75

90

1.000

519469.658

917469.02

901236.61

78
1000000.00

EP2, EP3 (MYR/year)

800000.00
600000.00
400000.00
200000.00
0.00
0

0.1

0.2

-200000.00

0.3

0.4

0.5

0.6

0.7

0.8

0.9

Jatropha conversion, XJ
EP2

EP3+recycle

EP3-recycle

Figure 2. 10: Graph of economic potential at the second level (EP2),

economic potential at the third level with recycle and economic potential
at the third level without recycle.

2.5
Comparison of Simulation (SuperPro) and Manual Calculation
Results
SuperPro version 6.0 had been used to run the simulation and
simulation result is compared to manual calculation result. Table 2.29 shows
the comparison of simulation and manual calculation results. The assumption
in this simulation are as follows;

Extent of reaction at fermenter is 100%

Efficiency of centrifuge and spray dryer is about 95%

Physical properties of Jatropha oil is assume similar to with physical

properties of soybean oil, which already registered in SuperPro data
bank.

Parameter such as temperature or pressure to be constant at input or

output

78

Table 2. 29: Comparison between Simulation and Manual Balance

EQUIPMENT
FERMENTER
FLAT BOTTOM
TANK
CENTRIFUGE 1
BLENDING TANK 1
MIXER
CENTRIFUGE 2
BLENDING TANK 2
CENTRIFUGE 3
SPRAY DRYER

INPUT
(kg/batch)
MANUAL
411.582

INPUT
(kg/batch)
SIMULATION
411.582

ERROR
(%)

OUPUT
(kg/batch)
SIMULATION
389.66

ERROR
(%)

0.0

OUTPUT
(kg/batch)
MANUAL
583.958

384.375

868.072

9.5

384.375

3127.685

69.35

384.375
758.75
1487.15
1855.1468
1831.0299
1794.4093
316.6493

554.546
758.756
3619.699
2183.448
9155.149
9233.36
5098.1

18.9
0.0
58.9
3.5
46.0
99.0
98.3

379.3781
743.575
1855.1468
1831.0299
1794.4093
1771.082
299.303

3247.263
1087.579
1515.433
78.214
9233.36
76.852
3789

82.92
49.28
40.35
98.38
64.82
98.99
99.70

36.47

Justification of Error: There is significant difference between manual and simulation result probably due to unregistered data of jatropha oil and
PHB, hence simulation cannot calculate the reaction kinetics. In the simulation, air flow rate is in default set to 100kg/batch per hour while
manual calculation is about 2200kg/hour needed for the whole process.

79
CHAPTER

UTILITIES & HEAT INTERGRATION

3.1

Introduction
Plant utilities are significant in any energy use or supply of services

from main centre of local state to plant such as electricity, steam and water to
complete the plant before plant operation. In economics, utility is a
representation of preferences over some set of goods and services. Preferences
have a (continuous) utility representation so long as they are transitive,
complete, and continuous. Utilities of the entire plant have been calculated in
this chapter to estimate the utility cost of the plant annually. Process of heat
integration is mainly refer to heat which was previously cooled off is
recovered and reused in another unit operation. Heat can be transferred from
one process stream to the other in a single heat exchanger by process of heat
integration. Justification of decision on heat integration was making based of
standard plant design.

3.2

Utilities

3.2.1

Electricity
Electricity in Peninsular Malaysia have been generated and supplied by

Tenaga Berhad (TNB) and this plant is charged based on standard electricity
charge which is 33.70sen/kWh. The table below show all the power
consumption of equipment used in plant design mainly pumps and blowers.
Table 3. 1: Total Power consumption of equipment used in plant design
Equipment

Power Consumption(kW)

P-1/G-101

172.790

P-12/PM-101

174.086

P-15/PM-102

174.086

P-17/PM-103

174.086

80
P-11/PV-105

10.58

P-14/PV-106

13.38

P-11/PV-105

10.58

Total usage

719.008

Total Cost(RM/year)

4797652.781

The total power usage in plant is 719.008 kWh. Average industrial

tariff for electricity from Tenaga Nasional Berhad is 33.70 sen /kWh . By
applying the industrial tariff of electricity 33.70 sen /kWh, the total electricity
cost per year is equal to RM 4797652.781/year with operation hours of 8000
per year. All the calculation is based on CEPCI 2014.

3.2.2

Steam
Steam is supply for sterilization purpose for killing the entire

microorganism in the media preparation tank, seed fermenters and bioreactors.

The cost of steam used is calculated according to standard steam tariff which
is $6.62/1000kg. All the steam supply needed is state in Table 3.2 below.
Table 3. 2: Total steam consumption of equipment used in plant design
Equipment

Steam supply (kg/h)

P-3/V-101

43.49

P-8/ST-101

187.20

P-7/ST-102

62.50

P-4/V-101

266.95

P-6/V-102

626.53

Total usage

1,186.67

Total Cost(RM/year)

9,140,191.175

The total steam usages for these main equipment are 1,186.67kg/h.
Calculate using the standard steam charges, the total steam cost is about RM
9,140,191.175 /year with operation hours of 8000 per year. By conversion, the
total steam cost is RM 9,140,191.175 /year.

81
3.2.3

Water
In Malaysia, state governments are responsible for the development,

operation and maintenance of water supplies. Entities for States Water Supply
Authorities in Malaysia are; Public Works Department, Water Supply
Department, Water Supply Board and Water Supply Company. According to
Syarikat Bekalan air Johor, the standard industrial water charges is RM
2.93/m. The main equipments which need supply of water is seed fermenters
and bioreactors.
Table 3. 3: Total water consumption of equipment used in plant design
Equipment

Water consumption (kg/batch)

P-5/SFR-101

113.840

P-4/V-101

418.062

P-6/V-102

3162.559

P-16/V-104

9155.050

Total usage

12,849.51

Total cost (RM/year)

7,454,515.37

The total water consumption for bioreactors and seed fermenters is

12,849.51 kg/batch. Through calculation the total cost of water is RM
7,454,515.37/year

year

with

operation

hours

8000

per

year.

RM

7454515.37/year is needed for water cost. By addition of total cost by

electricity, steam and water cost, the total cost of utilities is RM 4,797,652.781
+ RM 9,140,191.175 + RM 7,454,515.37 = RM 21,392,359.33 /year.

3.3

Heat Integration
Energy recovery is the main objectives of this design. Energy

integration is a technique to match hot and cold streams in a plant to achieve

heat transfer to reduce hot and cold utility consumption. The energy
consumption is a major part of plants operating cost. Generally, energy is
used for heating and cooling utilities. Before we proceed to design the heat

82
exchanger network, first, the base case process flow heat exchanger should be
known of its heat exchanger area and energy utilities consumption, in order to
compare it with the utility consumption after heat integration (Rossiter, 2010).
These energy utilities should be optimized in order to reduce the utilities
consumption of the process, which will affect the operating cost of this plant.
Heat exchanger network design and optimization can be a very tedious and
time-consuming task if the improper method is employed. Nevertheless, with
detailed planning, it can be a very rewarding step in reducing the design cost
and annual operating costs of the plant through a substantial reduction in the
energy requirements of a process.
Pinch technology is applied to determine the pinch temperature,
minimum heating and cooling requirements of the plant. Benefits of pinch
technology include:

Energy savings (15-90%)

Capital savings (up to 30%)
Cleaner processes (less fuels)
New design and retrofit
Simple and easy to use
Improved flexibility, reduce fouling problems
Pinch Analysis as the main methodology for determination of heat

integration. The most important new feature in Pinch Analysis was the ability
to establish Performance Targets ahead of design only based on information
about the change in thermodynamic state for the process streams. The most
important property of the Heat Recovery Pinch is that it decomposes the
process into a heat deficit region above Pinch and a heat surplus region below
Pinch(Sun & Luo, 2011). There is not enough heat available in the hot streams
above Pinch to satisfy the overall heating demand of the cold streams, and
external heating is required. Below Pinch, there is not enough cooling
available in the cold streams to satisfy the overall cooling demand of the hot
streams, and external cooling is required (Matija, 2002).
According to Pinch Technology Method (Matija, 2002) basic Heat
Exchanger Network Synthesis (HENS) must consist of:

83
1. A set of process stream to be cooled and a set of process stream to be
heated
2. The inlet and outlet flow rates and temperatures for all these process
streams
3. The heat capacities for each of the stream versus their temperature as
they pass through the heat exchanger process
4. The available utilities, their temperature and their costs per unit of heat
provided or removed

Heating and cooling utilities is a major part of the plants operating

costs. In order to increase the net profit of a plant, designer must focus on a
few solutions to reduce the usage of utilities to reducing the operating cost of
the whole plant. This can be done by maximizing the use of heating and
cooling streams generated by the plant itself. The process is called process
energy integration. Process integration can lead to a substantial reduction in
the energy requirements of a process, and consequently save up the operating
cost of the whole plant. Increasing of energy efficiency will solve the
problems of energy equilibrium system and reduce the cost of plant.
Since the overall process in the production of 50 MT of PHB did not
require any distillation column, cooling tower, boiler, condenser or heat
exchanger, the heat integration calculation is not required in this plant
designed. Plant for production of 50 MT PHB involve less temperature
differences in process flow, so there is no need of addition heating or cooling
equipment for heat exchange. According to Smith R (2012), addition of
unnecessary heat exchanger network at low temperature different will cause
uneconomical profit. Besides that, if low temperature different is used in heat
exchanger network such as lower than 100C, will cause high cost and
inadequacy effectiveness and accuracy to plant (Fraser & Hallale, 2000).

84
3.4

Economic Potential Level 5: Heat Integration System

In a typical process, there are normally several hot streams that must be

cooled and several cold streams that must be heated. The usage of external
cooling and heating utilities (e.g., cooling water, refrigerants, steam, heating
oils, etc.) to address all the heating and cooling duties is not cost effective.
Indeed, integration of heating and cooling tasks may lead to significant cost
reduction.
EP5 = EP4 Heat integration Cost
i.

EP4
Economic potential in level 4 is considering utility, recycle stream,

reactor cost and separation unit cost. The detail calculation is presented in
Chapter 4 and the calculated value is RM 169, 711/year.

ii.

Heat integration Cost

From the standard of plant design, for low temperature different in unit

operation process, there is no need addition of heat network exchanger as this

will bring high cost to plant design. So the cost for heat integration part can be
ignored.
EP5

= EP4 Heat integration Cost

= RM 169, 711/ year RM 0
= RM 169, 711/year

85
CHAPTER 4
EQUIPMENT SIZING

4.1

Introduction
Equipment sizing is where evaluation the specification of each of the

units used in the polyhdroxybutyrate (PHB) production plant from the type of
material used to the size of the units take place. All of the choices made is
based on the most suitable condition for the efficient and economic production
of PHB. This part is important as it enables the subsequent analysis that is
involved in process design such as economic analysis. According to Biegler,
(1997) the sizing will provide the data for the estimating the cost of the
equipment as the cost is increase nonlinearly with equipment size or capacity.
This PHB plant has a total of 12 types of equipment consists of seed
fermenter, main fermenter, gas compressor, air filter, heat sterilizer, splitter,
blending tank, storage tank, centrifuge, pumps, and spray dryer. The complete
summary result of all the equipment specifications are listed out in table form
and the detail sizing calculation is attached in Appendix C.

4.2

Heat Sterilizer (ST-101 & ST-102)

Table 4. 1: Sizing Summary of Heat Sterilizer
Equipment Specification Sheet

Item No.

ST-101 &ST-102
To heat process stream in order to prevent

Function

contamination

Material of construction

Carbon steel; floating head

Heater:
Heat Duty, Q (kJ)

1340.923 kJ

Log mean temperature, Tlm (oC)

2

Heat transfer area, A (m )

Cooler:

68.08
0.014

86
Heat Duty, Q (kJ)

-10399540.100

Log mean temperature, Tlm (oC)

39

Heat transfer area, A (m )

4.3

260.93157

Media Preparation Tank (P-09)

Table 4. 2: Sizing Summary of Media Preparation Tank

Equipment Specification Sheet

Item No.
Function
Material of Construction
Design Type
Tank Specification
Type of Culture
Diameter (m)
Height (m)
Working Volume (m3)
Tank Volume (m3)
Jacket Specification
Fluid
SIP Pressure (barg)
SIP Temperature (C)
SIP Duration (min)
Average Steam Flow Rate (kg/h)
Heat Transfer Area (m2)
Maximum Heat Capacity (kW)
Maximum Steam Flow Rate
(kg/h)

4.4

P-09
Raw material sterilization and storage
Stainless steel 316L
Vertical with hemispheral heads
Batch
0.70
1.40
0.53
0.62
Steam
1.1
130
30
40.02
0.34
26.27
43.49

Splitter (FSP-101 & FSP-102)

Table 4. 3: Sizing Summary of Splitter

Item No.
Volume, m3
Diameter, m
Height, m

Equipment Specification Sheet

FSP-101
0.2563
0.44
1.735

FSP-102
181.23
2.26
9.04

87
4.5

Gas Compressor (G-101)

Table 4. 4: Sizing Summary of Gas Compressor

Equipment Specification Sheet

Item No.
G-101
Type
Centrifugal / Turbine-driven
Material of construction
Carbon Steel
Operating Conditions
Inlet temperature, T1 (K)
298.15
Inlet pressure, P1 (bar)
1
Outlet pressure, P2 (bar)
4
Equipment sizing
Fluid flow rate (kg/batch)
22960.29
Horse power (hp)
19.31
80
Efficiency, c (%)

4.6

Air Filter (AF-101 & AF-102)

Table 4. 5: Sizing Summary of Air Filter

Equipment Specification Sheet

Pore size (m)
Diameter (mm)
Membrane
Membrane price (RM)
Depth of the filter membrane (m)
Cross sectional area (m2)

4.7

0.2
47
PTFE
9.22
0.8591
2.789

Seed Fermenter (V-101)

Table 4.1: Sizing Summary of Seed Fermenter

Equipment Specification Sheet

Item No.
P-5/V101
Function
Cell Culture
Material of Construction
Stainless steel 316L
Design Type
Vertical with hemispheral heads
Operating Temperature (oC)
37
Operating Pressure (atm)
1
Aeration rate (vvm)
3
Duration (h)
24
Tank Specification

88
Type of Culture
Diameter (m)
Height (m)
Working Volume (m3)
Tank Volume (m3)

Batch
0.30
0.60
1.123
1.404
Impeller Specification
Type of Impeller
Rushton Turbine
No of Impeller
4
No of Blades
6
Rotation Speed (rpm)
150
Diameter (m)
0.100
Width (m)
0.020
Length (m)
0.025
Duration (h)
24
Baffles Specification
No of Baffles
4
Diameter (m)
0.03
Height (m)
0.55
Baffles Clearance (m)
0.10
Heating Jacket Specification
Fluid
Steam
SIP Pressure (barg)
1.1
o
SIP Temperature ( C)
130
SIP Duration (min)
30
Average Steam Flow Rate (kg/h)
266.95
2
Heat Transfer Area (m )
2.50
Maximum Heat Capacity (kW)
177.37
Maximum Steam Flow Rate (kg/h)
293.65

89
4.8

Main Fermenter (V-103)

Table 4.7: Sizing Summary of Main Fermenter

Equipment Specification Sheet

Item No.
P-7/V103
Function
Fermentation
Material of Construction
Stainless steel 316L
Design Type
Vertical with hemispheral heads
Operating Temperature (C)
30
Operating Pressure (atm)
1
Aeration rate (vvm)
3
Duration (h)
24
Tank Specification
Type of Culture
Fed Batch
Diameter (m)
2.3032
Height (m)
4.6064
Working Volume (m3)
20.471
3
Tank Volume (m )
25.589
Impeller Specification
Type of Impeller
Rushton Turbine
No of Impeller
4
No of Blades
6
Rotation Speed (rpm)
150
Diameter (m)
0.7677
Width (m)
0.1536
Length (m)
0.1919
Baffles Specification
No of Baffles
4
Diameter (m)
0.23032
Height (m)
4.6065
Baffles Clearance (m)
0.7677
Heating Jacket Specification
Fluid
Steam
SIP Pressure (barg)
1.1
o
SIP Temperature ( C)
130
SIP Duration (min)
30
Average Steam Flow Rate (kg/h)
266.95
Heat Transfer Area (m2)
33.75
Maximum Heat Capacity (kW)
378.43
Maximum Steam Flow Rate (kg/h)
626.53

90
4.9

Storage Tank (V-104)

Table 4.2: Sizing Summary of Storage Tank

Equipment Specification Sheet

Item No.
V-104
Function
storage
Material of Construction
Stainless steel 316L
Design Type
Vertical with hemispheral heads
Tank Specification
Type of Culture
Batch
Diameter (m)
0.75
Height (m)
1.40
3
Working Volume (m )
0. 70
Tank Volume (m3)
0.90

4.10

Centrifuge (DS-101, DS-102 & DS-103)

Table 4.3: Sizing Summary of Centrifuge

Item No.

Equipment Specification Sheet

DS-101
DS-102

DS-103

Function

Cell biomass
separation

Product and
cell debris
separation

Product and
chemical
separation

Material of
Construction
Design Type

Carbon steel

Carbon steel

Carbon steel

Disc stack
Disc stack
Centrifuge Specification

Disc stack

Outer Diameter (m)

0.33

0.33

0.33

Inner Diameter (m)

0.16

0.16

0.16

Minimum Disc

63

58

11

Angle between Disc ()

45

45

45

Rotation Speed (rpm)

100

450

7500

Sigma Factor (m2)

29.1407

24.4627

334.6667

91
4.11

Blending Tank (V-105 & V-106)

Table 4.4: Sizing Summary of Blending Tank
Equipment Specification Sheet
V-105
V-106

Item No.
Function

Cell disruption

Blending

Material of
Construction

Carbon steel

Carbon steel

Design Type

Vertical with
hemispheral heads

Vertical with
hemispheral heads

Type of Culture
Diameter (m)
Height (m)
Working Volume (m3)
Tank Volume (m3)
Type of Impeller
Diameter (m)
Width (m)
Length (m)
Rotation Speed (rpm)
Motor Power (kW)

4.11

Tank Specification
Batch
0.81
1.62
0.89
1.12
Impeller Specification
Rushton Turbine
0.27
0.05
0.06
92.71
10.58

Batch
0.98
1.96
1.57
1.96
Rushton Turbine
0.27
0.05
0.06
92.71
10.58

Pumps
Table 4.5: Sizing Summary of Pumps

Equipment Specification Sheet

Identification
PM-101
PM-102
Type
Centrifugal
Centrifugal
Materials of construction
Cast iron
Cast iron
Pump Specification
50
50
Pump Efficiency, p (%)
Mechanical Efficiency, m
90
90
(%)
Horsepower (hp)
88.76
142.02

Other Pumps
Centrifugal
Cast iron
50
90
0.5

92
4.12

Spray Dryer (SDR-101)

Table 4.6: Sizing Summary of Spray Dryer

Equipment Specifications Sheets

Item No.
SDR-101
Function
To produce a dry powder
Materials of Construction
Carbon steel
Spray Dryer Specification
Discharge diameter of nozzle nipple (mm) 1.5627
Optimum pressure of the slip (atm)
8.04-8.93
Output of nozzle in (liters/h)
364.32
Diameter of jet of sprayed slip (m)
2.0091
Height of jet (m)
2.5896
Diameter of the drying chamber (m)
2.1431
Height of cylindrical part of drying 2.6789
chamber (m)
Height of conical part (m)
1.5627
Total internal height of drying chamber 4.2415
(m)
Volume of drying chamber (m3)
58.934

4.13

Economic Potential Level 4 (EP4): Separation System

4.13.1 General Structure of the Separation System

The composition of separation feed is that of the reactor effluent and
the separator section is to produce a final product of acceptable purity and a
stream of by-products. In this PHB plant separation system, centrifugation
plays an important role in the separation of the desired components from waste
in the streams, such as first centrifuge unit is function in cell biomass
separation process, second centrifuge unit is used to separate cell debris and
PHB, while for the third centrifuge is purpose in PHB and chemical impurities
separation. From the downstream process, there are three centrifugation units
needed to complete the separation process. Economic potential 4 was
evaluated by using following formula;
EP4 = EP3 Separation Unit Cost

93
4.13.1.1

EP3

Economic potential in level 3 is considering utility, recycle stream and

reactor cost. The detail calculation is presented in previous section and the
calculated value is RM 917,469.00/year.

4.13.1.2

Separation Unit Cost

From the calculation of centrifuge sizing, the disc diameter of

centrifuge was 0.5m. By assuming the diameter of the centrifuge disc is the
same with the different number of disc required and the rotation speed, the
purchased cost of the centrifuge is obtained. The index of 2014 was 574. Table
below shows the bare module cost for three centrifuges in PHB plant.
Table 4. 6: Bare Module Cost (CBM) for Centrifuges
Equipment
Centrifuge DS-101
Centrifuge DS-102
Centrifuge DS-103
Total

EP4

= EP3 Separation Unit Cost

= RM 917,469.00 RM 747,757.33
= RM169, 711/year

CBM (MYR)
12,475.32
72,194.35
663,087.65
747,757.33

94
CHAPTER 5
PROCESS CONTROL & SAFETY

5.1

Introduction
According to Institute of Instrumentation and Control, a piping and

instrumentation diagram (P&ID) is defined as a diagram which shows the

interconnection of process equipment and the instrumentation used to control
the process. In the process industry, a standard set of symbols is used to
prepare drawings of processes and as the primary schematic drawing used for
laying out a process control installation. P&ID plays a significant role in the
maintenance, modification, as well as regulation of the process that it
describes. It is critical to demonstrate the physical sequence of equipment and
systems, as well as how these systems connect. During the design stage, the
diagram also provides the basis for the development of system control
schemes, allowing for further safety and operational investigations, such as
Hazard and Operability Study. P&ID and its justification for this PHB plant is
laid out in Appendix D.3.
This chapter will emphasize on safety study of this PHB production
plant which includes identification of hazard and the risk assessment through
material safety data sheet and hazard and operability studies.
Major equipment control also will be covered as it is important to
regulate process conditions such as flow rate, pressure and temperature in
order to obtain stable, safe and healthy operations. Quality products also can
be produced efficiently and economically.

5.2

Identification of Hazard
Hazard identification procedure is used to identify the types of adverse

health effects that can be caused by exposure to some agent in question, and to
characterize the quality and weight of evidence supporting this identification.
Risk assessment includes determination of the events that can produce an

95
accident, the probability of those events, and consequences that could include
human injury or loss of life, damage to the environment, or loss of production
and capital equipment. Hazard identification can be performed independent of
risk assessment, but it would obtain best result if they are done together.
Figure 5.1 shows hazard identification and risk assessment procedure.

Figure 5. 1: Procedure of hazard identification and risk assessment.

(Source: Guidelines for Hazards Evaluation Procedures: American
Institute of Chemical Engineers, 1985)

5.2.1

Material Safety Data Sheet

The Material Safety Data Sheet (MSDS) is a detailed information

bulletin as a general guideline about PHB that describes the physical and

96
chemical properties, important characteristics, hazards/symptoms, preventive
measures, fire extinguishing/first aid, spillage and labeling of a chemical. The
precise format of an MSDS is not presently defined by regulation but there are
some minimum requirements. The minimum requirements for MSDS
according to Turton et al. (2009) are as follows;
1. An MSDS is required for each hazardous chemical, any chemicals
assigned a Threshold Limit Value (TLV), or any material
determined to be cancer causing, corrosive, and toxic, an irritant, a
sensitizer, or one that has damaging effects on specific body
organs.
2. Written in English.
3. Identity used on label.
4. Chemical name and common name of all ingredients that are
hazardous and that are present in 1% concentration or that could
be released in harmful concentrations.
5. Chemical name and common name of all ingredients that are
carcinogens and that are present in 0.1% concentration or that
could be released in harmful concentrations.
6. Physical and chemical characteristics of the hazardous chemical.
7. Physical hazards of the hazardous chemical, including the potential
for fire, explosion, and reactivity.
8. Health hazards of the hazardous chemical, including signs and
symptoms of exposure, and any medical conditions that are
generally recognized as being aggravated by exposure to the
chemical.
9. Primary routes of entry.
10. Occupational

Safety

and

Health

Administration

(OSHA)

permissible exposure limit, American Conference of Governmental

Industrial Hygienists (ACGIH) TLV, and any other exposure limit
used or recommended by the chemical manufacturer, importer, or
employer.
11. Whether the hazardous chemical is listed in the National
Toxicology Program (NTP) Annual Report on Carcinogens or has

97
been found to be a potential carcinogen in the International Agency
for Research on Cancer (IARC) Monographs, or by OSHA.
12. Any generally applicable precautions for safe handling and use that
are known to the chemical manufacturer, importer, or employer
preparing the MSDS, including appropriate hygienic practices,
protective measures during repair and maintenance of contaminated
equipment, and procedures for cleanup of spills and leaks.
13. Any generally applicable control measures that are known to the
chemical manufacturer, importer, or employer preparing MSDS,
such as appropriate engineering controls, work practices, or
personal protective equipment.
14. Emergency and first aid procedures.
15. Date of preparation of the MSDS or the last change to it.
16. Name,

address,

and

telephone

number

of

the

chemical

manufacturer, importer, employer, or other responsible party

preparing of distributing the material safety data sheet that can
provide additional information on the hazardous chemical and
appropriate emergency procedures, if necessary.

This information helps to prepare employers as well as employees to

respond effectively to daily exposure situations as well as to emergency
situations. The production of PHB in this plant is by using fermentation of
Cupriavidus necator with jatropha oil. The chemical used in this plant are
urea, Triton X-100 surfactant as well as sodium hypochlorite. The related data
for these chemicals can be found in Appendix D.1.

5.2.2

DOW Fire and Explosion Index

DOW Fire and Explosion Index (FEI) is one of the popular hazard

surveys developed by the Dow Chemical Company and published by the

American Institute of Chemical Engineers in the 1960s and is today used by
many companies to identify high-risk systems (Turton et al., 2004). It is a
formal systematized approaches using a rating form, and the final rating

98
number provides a relative ranking of the hazard. Figure 5.2 shows general
steps in calculating DOW Fire and Explosion Index but this chapter will
emphasize until FEI only. It starts with determining the material factor (MF)
that is a function only of type of chemicals used. The MF is the basic starting
value in the computation of the FEI and other risk analysis values. The factor
is ranged from 0 to 60 that indicates the magnitude of the energy release in a
fire or explosion. For non-combustible materials, the factor is zero while for
the combustible materials, the factor can be calculated from the following
equation:

where

is heat of combustion. The factor is then adjusted for general and

special process hazards based on conditions such as storage above the flash or
boiling point, endothermic or exothermic reaction, and fired heaters (Crowl
and Louvar, 2002). In general the higher the value of material factor, the more
flammable and or explosive the material. Figure 5.3 shows form used in the
DOW Fire and Explosion Index

99

Figure 5. 2: General steps in determining DOW Fire and Explosion Index

100

Figure 5. 3: Form used in DOW Fire and Explosion Index

The numerical value of the Process Unit Hazards Factor is determined

by first determining the General Process Hazards Factor (F1) and Special
Process Hazards Factor (F2) listed on the F&EI form. Each item which
contributes to the Process Hazards Factors contributes to the development or
escalation of an incident that could cause a fire or an explosion. General
Process Hazards (F1) covers six items;

101

Exothermic chemical reactions

The penalty varies from 0.3 for a mild exothermic, such as

hydrogenation, to 1.25 for a particularly sensitive exothermic, such as

nitration.

Endothermic processes
A penalty of 0.2 is applied to reactors, only. It is increased to 0.4 if the

reactor is heated by the combustion of a fuel.

Materials handling and transfer

This penalty takes account of the hazard involved in the handling,

transfer and warehousing of the material.

Enclosed or indoor process units

Accounts for the additional hazard where ventilation is restricted.

Access of emergency equipment

Areas not having adequate access are penalized. Minimum requirement

is access from two sides.

Drainage and spill control

Penalizes design conditions that would cause large spills of flammable

material adjacent to process equipment; such as inadequate design of drainage.

These factors are intended to allow for the general process hazards
associated with the unit being considered. The Special Process Hazards (F2)
are factors that are known from experience to contribute to the probability of
an incident involving loss. Twelve factors are listed in the calculation form
based on Figure 5.3;

Toxic materials
The presence of toxic substances after an incident will make the task of

the emergency personnel more difficult. The factor applied ranges from 0 for
non-toxic materials, to 0.8 for substances that can cause death after short
exposure.

Sub-atmospheric pressure

102
Allows for the hazard of air leakage into equipment. It is only applied
for pressure less than 500 mmHg (9.5 bar).

Operation in or near flammable range

Covers for the possibility of air mixing with material in equipment or

storage tanks, under conditions where the mixture will be within the explosive
range.

Dust explosion
Covers for the possibility of a dust explosion. The degree of risk is

largely determined by the particle size. The penalty factor varies from 0.25 for
particles above 175 m, to 2.0 for particles below 75 m.

Relief pressure
This penalty accounts for the effect of pressure on the rate of leakage,

should a leak occur. Equipment design and operation becomes more critical as
the operating pressure is increased. The factor to apply depends on the relief
device setting and the physical nature of the process material.

Low temperature
This factor allows for the possibility of brittle fracture occurring in

carbon steel, or other metals, at low temperatures.

Quantity of flammable material

The potential loss will be greater the greater the quantity of hazardous

material in the process or in storage. The factor to apply depends on the

physical state and hazardous nature of the process material, and the quantity of
material. It varies from 0.1 to 3.0 and 5 in the DOW Guide.

Corrosion and erosion

Despite good design and materials selection, some corrosion problems

may arise, both internally and externally. The factor to be applied depends on
the anticipated corrosion rate. The severest factor is applied if stress corrosion
cracking is likely to occur.

Leakage joints and packing

103
This factor accounts for the possibility of leakage from gaskets, pump
and other shaft seals, and packed glands. The factor varies from 0.1 where
there is the possibility of minor leaks, to 1.5 for processes that have sight
glasses, bellows or other expansion joints.

Use of fired heaters

The presence of boilers or furnaces, heated by the combustion of fuels,

increases the probability of ignition should a leak of flammable material occur

from a process unit. The risk involved will depend on the siting of the fired
equipment and the flash point of the process material. The factor to apply is
determined with reference to Figure 6 in the Dow Guide.

Hot oil heat exchange system

Most special heat exchange fluids are flammable and are often used

above their flash points; so their use in a unit increases the risk of fire or
explosion. The factor to apply depends on the quantity and whether the fluid is
above or below its flash point

Rotating equipment
This factor accounts for the hazard arising from the use of large pieces

of rotating equipment: compressors, centrifuges, and some mixers..

The computation of DOW FEI completed in Risk Analysis Summary
to determine the degree of hazard. Below is the table for determining the
degree of hazard from the Dow Fire and Explosion Index based on Crowl and
Louvar (2002).

Table 5. 1: Degree of Hazard based on DOW Fire and Explosion Index

(FEI)
DOW Fire and Explosion Index

Degree of Hazard

1-60

Light

61-96

Moderate

97-127

Intermediate

128-158

Heavy

104
Severe

159 and above

Calculation of Dow Fire and Explosion Index for urea, Triton X-100
surfactant, and sodium hypochlorite are shown in Appendix D.1.

5.2.2

Toxicity
According to Crowl and Louvar (2002), toxicity of a chemical or

physical agent is a property of the agent describing its effect on biological

organisms. Table 5.2 shows subdivision of toxicity and its respective
explanation.
Table 5. 2: Toxicity level
Level

Explanation

Acute

Adverse effects are observed within a short time of exposure to

the chemical. This exposure may be a single dose, or a short
continuous exposure, or multiple doses administered over 24
hours or less.

Sub-

Adverse effects are observed following repeated daily exposure to

acute

a chemical, or exposure for a significant part of an organisms

lifespan (usually not exceeding 10%). With experimental animals,
the period of exposure may range from a few days to 6 months.

Chronic

Adverse effects are observed following repeated exposure to a

chemical during a substantial fraction of an organisms lifespan
(usually more than 50%). For humans, chronic exposure typically
means several decades; for experimental animals, it is typically
more than 3 months. Chronic exposure to chemicals over periods
of 2 years using rats or mice may be used to assess the
carcinogenic potential of chemicals.

105
Indicators of toxicity hazards include LD50, LC50, plus a wide range
of in vitro and in vivo techniques for assessment of skin and eye irritation, skin
sensitization, mutagenicity, acute and chronic dermal and inhalation toxicity,
reproductive toxicology, carcinogenicity etc.
The LD50 is the statistically derived single dosage of a substance that
can be expected to cause death in 50% of the sample population. It is therefore
an indicator of acute toxicity, usually determined by ingestion using rats or
mice, although other animals may be used. LD50 is also determined by other
routes, e.g. by skin absorption in rabbits. The values are affected by species,
sex, age, etc. The LC50 is the lethal concentration of chemical (e.g. in air or
water) that will cause the death of 50% of the sample population. This is most
appropriate as an indicator of the acute toxicity of chemicals in air breathed (or
in water, for aquatic organisms). Table 5.3 illustrates the use of LD50 values
to rank the toxicity of substances.
Table 5. 3: Toxicity rating system

Toxicity
Rating

Commonly
Used Term

Extremely
Toxic
Highly
Toxic
Moderately
Toxic
Slightly
Toxic
Practically
non-toxic
Relatively
harmless

2
3
4
5
6

LD50
Single
Oral
Dose for
Rats
(g/kg)
0.001

4hr Vapor
Exposure
Causing 2 to
4 Deaths in 6
Rat group
(ppm)
<10

0.0010.05
0.05-0.5

10-100

0.5-5.0

1000-10000

5.0-15.0

10000100000
>100000

>15.0

100-1000

LD50
Skin for
Rabbits
(g/kg)
0.005
0.0050.043
0.0440.340
0.35-2.81
2.82-22.6
>22.6

Probable
Lethal
Dose for
Humans
Taste (1
grain)
1 teaspoon
(4ml)
1oz (30g)
1 pint
(250g)
1 quart
(500g)
>1 quart

Toxicity index of chemical substances used in this 50 MT PHB

production is attached in Appendix D.1.

106
5.3

Hazard and Operability Studies (HAZOP) of Major Equipment

Hazard and Operability Studies or simply known as HAZOP is a

formal procedure to identify hazards in chemical and biochemical process

facility (Guidelines for Hazard Evaluation Procedures, 3rd ed.). The procedure
is considered effective in identifying hazards and is well accepted by chemical
and biochemical industries (Crowl and Louvar, 2011).
The basic procedure is to assemble a carefully chosen team of
individuals and systematically search through P & I Diagrams to identify the
possible deviation and its causes as well as its consequences. The structure of
the search is provided by the use of simple word models to create potential
deviations at each point in the plant. Then, it will be decided whether the
search deviation can be applicable or not. Actions for prevention and
mitigation of the consequences will be provided. Basically, one of the concept
that the group must bear in mind that HAZOP studies should not be used to
solve problems, it is a proactive method, to identify and assess the impact of
the hazards.
The HAZOP technique applies to combination of a Guide Word to
generate a Deviation from design intent. This technique is systematically
applied to parts of system such that hazard and operability problems on
complete system are eventually identified. Table 5.4 shows guide words used
for HAZOP procedures.
Table 5. 4: General Guide Words for HAZOP procedures (Crowl and
Louvar, 2002)
Guide Words
NO,NOT,NONE

Meaning
The complete
negation of the
intention
MORE,HIGHER,GREATER Quantitative
increase

LESS,LOWER

Quantitative
decrease

Comments
No part of the design
intention is achieved,
nothing else happens
Applies to quantities such as
flow rate and temperature
and to activities such as
heating and reaction.
Applies to quantities such as
flow rate and temperature
and to activities such as
heating and reaction.

107
AS WELL AS

Quantitative
increase

PART OF

Qualitative
decrease

REVERSE

The logical
opposite of

OTHER THAN

Complete
substitution

SOONER THAN

Too early or in
the wrong
order
Too late or in
the wrong
order
In additional
locations

LATER THAN

WHERE ELSE

All the design and operating

intentions are achieved along
with some additional activity
such as contamination of
process stream.
Only some of the design
intentions are achieved,
some are not.
Most applicable to activities
such as flow or chemical
reaction. Also applicable to
substances, for example,
poison instead of antidote.
No part of the original
intention is achieved; the
original intention is replaced
by something else.
Applies to process steps or
actions.
Applies to process steps or
actions.
Applies to process locations,
or locations in operating
procedures.

The HAZOP procedure uses the following steps to complete an analysis:

1. Begin with a detailed flow sheet and break the flow sheet into a
number of process units.
2. Select a unit (a fermenter for example) and choose a study node
(i.e vessel, line)
3. Describe the design intent of the study node. For example,
fermenter is designed to be place for biochemical takes place
and produces desired product.
4. Select a process parameter (i.e flow, temperature, pressure)
5. Apply guide word to the process parameter to suggest possible
deviations
6. If the deviation is applicable, determine possible causes and
note any protective systems.

108
7. Evaluate the consequences of the deviation (if any)
8. Recommend action.
9. Record all information.

For this 50 MT production of (PHB) plant design, only major

equipment such as gas compressor, fermenter, blending tank, and spray dryer
will be evaluated for HAZOP study and can be found in Appendix D.2.

5.4

Major Equipment Control

Process control has become increasingly important in the process

industries as a consequence of global competition, rapidly changing economic

conditions, faster product development, and more stringent environmental and
safety regulation (Seborg et al., 2011). Generally five elements are controlled
in the plant that is temperature, pressure, flow rate, level in the equipment and
composition. There are four basic components of a control system;
i.

sensor (primary element),

ii.

transmitter (secondary element),

iii.

controller (brain of the control system), and

iv.

final control element (such as control valve,

v.

other elements are variable speed pumps, conveyors and electric

motors).

The control of a process is often accomplished by measuring the

variables (controlled variables), comparing this measurement with the value at
which it is desired to maintain the controlled variables (set point), and
adjusting some further variables (manipulated variables) which has a direct or
indirect effect on the controlled variables.
There are two types of control system that is a feedback control and a
feed forward control. In feedback control system, it is applied when measuring
device detected the output of a process and then a controlling device will be
compared between the measured reading and the process set point, and lastly
the signal will be sent to the final control element that will manipulate the

109
controlled variables. For feedback control, the disturbance variable is not
measured. Anyhow, a feed forward control configuration measures the
disturbances (load) directly and takes control action to eliminate its impact on
the process output. That is mean a feed forward controllers have the theoretical
potential for perfect control.

5.4.1

Objectives of Control System

This plant consist five major equipment. There are seed fermenter,

fermenter, blending storage, disk stack centrifuge and spray dryer.

Each

equipment has their own hazard. Below is the equipment process control for
each equipment. The primary objectives of the instrumentation and control
schemes are to maintain a process at the desired operating conditions, safety
and efficiently, while satisfying environmental and product quality
requirement;
(i) Safety of plant operation:
-

To keep the process variables within known safe operating

limits

To detect dangerous situations as they develop and to provide

alarms and automatic shut-down system.

(ii) Production rate

-

To get the desired amount and the quality of the final product.

To maintain the product composition within the specified

quality standard is essential.

(iii) Operating Constraint

-

Various types of equipment used in this plant have operation

constraints inherent to their operation. Such constraint should
be satisfied through operation of plant. For example, the
distillation column should not be flooded; temperature in a
reactor should not exceed upper limit; tanks should not

110
overflow or go dry; pumps must be maintained at a certain net
positive suction head.
-

A control system should be set up to satisfy all these

operational constraints.

(iv) Economic
-

To operate at the lowest production cost

The operation of the plant must conform to the market

condition, which is availability of raw materials and demand of
the final product.

(v) Environmental Regulations

-

Variable controlled must not exceed the allowable limits set by

laws.

5.4.2

Process Control of Major Equipment

This section provides summary of process control summary of major

equipment in 50 MTPA PHB plant comprises of seed fermenter, main

fermenter, blending storage, disk stack centrifuge and spray dryer.

5.4.2.1 Seed Fermenter (V-101)

The control system for the seed fermenter, V-101 is designed so that
this unit can be operated optimally and safely. The following are the control
objectives of a seed fermenter:
a. Control the culture medium level to suppress flooding or to avoid
the culture medium level in the fermenter from being too low.
b. Control the pH point of the culture medium at a desired value for
healthy Cupriavidus necator H16 growth
c. Control the pressure in the fermenter at a desired value for healthy
Cupriavidus necator H16 growth.

111
d. Control the temperature of the culture medium at a constant
temperature to counteract the heat generated by aerobic oxidation.

Figure 5. 4: Section of P & ID of Seed Fermenter

Controlled
Variable
Flow

Manipulated
Variable
Inlet flow rate

Disturbance
Outlet flow rate

Type of
Controller
Feedback

5.4.2.2 Main Fermenter (V-102)

The control system for the main fermenter, V-102 is designed so that
this unit can be operated optimally and safely. The following are the control
objectives of a seed fermenter:
a. Control the culture medium level to suppress flooding or to avoid
the culture medium level in the fermenter from being too low.
b. Control the pH point of the culture medium at a desired value for
healthy Cupriavidus necator H16 growth
c. Control the pressure in the fermenter at a desired value for healthy
Cupriavidus necator H16 growth.
d. Control the temperature of the culture medium at a constant
temperature to counteract the heat generated by aerobic oxidation.

112

Figure 5. 5: Section of P & ID of Main Fermenter

Controlled

Manipulated

Variable

Variable

Flow

Inlet flow rate

Disturbance
Outlet flow rate

Type of
Controller
Feedback

5.4.2.3 Disc Stack Centrifuge (DS-101, DS-102, DS-103)

Figure 5. 6: Section of P & ID of Disc Stack Centrifuges

Controlled

Manipulated

Variable

Variable

Disturbance

Type of
Controller
Programmable

Flow

Inlet flow rate

Outlet flow rate

Logic Controller
(PLC)

113
5.4.2.5 Spray Dryer

Figure 5. 7: Section of P & ID of Spray Dryer

Controlled
Variable
Flow

5.5

Manipulated
Variable
Inlet flow rate

Disturbance
Outlet flow rate

Type of
Controller
Feedback

Piping and Instrumentation Diagram

Piping and Instrumentation Diagram (P&ID) for the whole process of

PHB plant has been constructed accordingly and is attached in Appendix D.3.

114
CHAPTER 6
WASTE MANAGEMENT AND POLUTION CONTROL

6.1

Introduction
Waste management is the collection, transportation and disposal of

garbage, sewage and other waste products. Waste management encompasses

management of all processes and resources for proper handling of waste
materials, from maintenance of waste transport trucks and dumping
facilities to compliance with health codes and environmental regulations.
The waste management hierarchy is a nationally and internationally
accepted guide for prioritizing waste management practices with the objective
of achieving optimal environmental outcomes. It sets out the preferred order of
waste management practices, from most to least preferred.
The waste management hierarchy is one of the guiding principles of
the Zero Waste SA Act 2004, and is regarded in South Australias Waste
Strategy 2011 2015 as a key element for guiding waste management
practices in South Australia, while still recognizing the need for flexibility
based on local and regional economic, social and environmental conditions.

Figure 6. 1: Waste management hierarchy

115
6.1.1

Higher Up the Hierarchy

The further activity moves up the waste management hierarchy, the

more greenhouse gains there are to be made. Reuse requires less energy than
recycling, although designs which are both adaptable and durable are essential
to its success. Other factors, such as the consumer desire for newness, can
conspire against reuse. There are many ways that clothes, cars, books,
buildings and other materials are currently reused, such as:

trash and treasure markets

e-bay

free giveaway swap websites.

Reuse is already part of our society, so there is an existing precedent to

build on. Reduce requires less energy again, by designing out waste before it
is created. Waste, in all its guises, is an indicator that systems and processes
could be designed better. It makes no sense to pay both financial and
energy/greenhouse costs for waste twice first to create it, then to dispose of
it. Avoid is the ultimate zero waste challenge; the highest point on the
hierarchy. The volume and rate at which resources are being channeled
through the human economy needs to be slowed, along with a recognition that
all our material goods have an energy 'price tag'.
To effectively address the zero waste and climate change agenda, there
needs to be a move beyond recycling into the largely uncharted territory of the
higher end of the hierarchy, to reuse, reduce and avoid, with a particular
emphasis on eco-efficiency (the same or greater utility from less material
input).
As chemical industry grows rapidly, several problems have arises.
Therefore, terms such as well-developed waste minimization plan and
Environmental Safety Management Program are heard every time a chemical
plant is set up. There is a standard known globally as ISO 14000. ISO 14000
standard is mainly to ensure waste is minimized from a chemical plant and to
provide a better living for the workers and people living near the plant.

116
Waste treatment may seem to be an economic disadvantage for
processing plant because more money has to be spent on the waste treatment
process. This kind of prospective is wrong. An engineer must be more ethical
in order to ensure our environment is protected in the long run.
As any other chemical plant, polyhdroxybutyrate process plant also
produces wastes in liquids and gaseous form. The main sources of waste are in
liquid form comes from air filter (AF-101 and AF-102) consisting wastewater.
Both the main fermenter and seed fermenter (V-101 and -102) produce CO2
and N2 as waste. This main waste still contain an appreciable quantity of
organic substances and therefore must be purified before it is discharged from
the plant.

6.1.2

Waste Minimization
The best way to minimize waste is through source reduction. Source

reduction is a way to reduce the production of waste in the process. It is better

not to produce waste rather than to implement extensive treatment schemes to
insure that the quality of the environment is not damaged. Source reduction
can accomplish by:
i.

Good housekeeping practice

ii.

Input material modification

iii.

Technology modification

Technology modification can be further categorize as follows:

1. Process modification.
2. Improved control.
3. Equipment changes.
4. Energy conservation.
5. Water conservation

117
6.1.3

Objective of Waste Minimization

Minimization of waste is the step taken before implementing waste

treatment. By doing so, manufacturers can save a lot of money by reducing

waste treatment and disposal costs.

The purchase of raw material and

operating costs will also be reduced.

The step above also helps manufacturers to meet national policy goals
that are to protect the environment and promote good worker and public
health. In this chapter we will not discuss more about this step, as we are
more interested in treating the waste.

6.1.4

Waste Sources and Effect to Human and Environment

The waste from the plant contains various types of chemical

compounds. This chemical compounds must be ensured to comply with the

Malaysia Environmental Quality Act 1974 before discharged to the
environment.
Before the wastes are discharged to the environment, some
consideration must be done. Firstly, we have to consider the economic aspect,
whether the waste can be recover and sell as a product or not. Secondly, we
have to consider the waste properties, whether it can be discharged directly to
the environment. Thirdly, we have to consider from the safety reason, whether
it is dangerous to the environment or not.
Specifically there are several equipment that contain waste material in
this process. The source of these waste generation is shown as below and the
amount of each waste produced is described as below:
Table 6. 1: Source and Waste Generated in PHB plant
Source

Waste

Seed fermenter V-101

Carbon dioxide

Main fermenter V-102

Carbon dioxide

Centrifuge DS-101

Biomas, nutrient medium

118
Centrifuge DS-102

Water, biomass, surfactant (Triton X100), hypochlorite

Centrifuge DS-103

Water, biomass, surfactant (Triton X100), hypochlorite

Spray dryer SDR-101

Water, surfactant (Triton X-100),

hypochlorite

Based on the table shown above, it is clear that seed fermenter and also
fermenter used in the production of 50 MT of PHB per year will emit carbon
dioxide gas as the by-product of the fermentation reaction that took place.
From the mass balance calculation, total amount of carbon dioxide emission
from seed fermenter V-101 and main fermenter V-102 are 31.99kg/batch. For
1st centrifuge, wastewater generated consists of biomass and nutrient medium
that is 353.54 kg/batch. In the second centrifuge, wastewater consist of 635.98
kg/batch of residual liquid produced in that centrifuge, 619.9615 kg/hr
constitutes of cell debris, 0.08935 kg/batch of biomass while water of 15.931
kg/batch. Meanwhile, in the third centrifuge, mass balance calculation
revealed that 5589.20 kg/batch of wastewater is generated from the
centrifugation.

6.1.5

Waste Management Option for Each Waste Produced

Table 6. 2: Waste Management Options by Our Company
Type of Waste

Waste Management Option

Carbon dioxide gas

Direct emission to environment

Biomass

Sell to fertilizer company : Sarawak

Fertilizer Sdn Bhd

Sodium hypochlorite

Subjected to filtration using activated

carbon filtration system and treatment by
Kualiti Alam

Water + Surfactant

Treatment using activated sludge system.

119
Carbon dioxide gas is the by- product of fermentation reaction that
took place in the seed fermenter and fermenter in our plant. We employed only
one fermenter for our PHB production, where a total of 10244.52 kg/year of
this gaseous residue will be emitted. Initially, the gases will be filtered by air
filter installed in those seed fermenter and fermenter before it is discharged
directly to the environment. This step is very essential in order to prevent any
escape of microorganisms from the reactor. Hence, the safety level of the
emitted gas is very high and it is totally free from any risk of microorganisms
escape. No further treatment is applied to this gas since the concentration of
carbon dioxide gas released is below the discharge limit. The reference used
for comparison of the concentration of carbon dioxide used is The Malaysian
Standard Guidelines for Air Gaseous Pollutants under EQA as shown in Table

Table 6. 3: Malaysian Standard Guidelines for Air Gaseous Pollutants

Pollutant

Averaging

Malaysian

Malaysian

Time

Guideline

Guideline

(ppm)

(g/m3)

Compliance

CO2

8 hour

5000

90

1997

Particles

24 hours

260

1995

I year

90

Pollutant

Averaging time

Guidelines, g/cm3

Malaysia Ambient Air Quality Guidelines

Particulate Matter (PM10)

24h

150

Total Suspended Particulate (TSP)

24h

260

24h

30

Canada
Particulate Matter (PM2.5)

United States of America

Particulate Matter (PM10)

24h

150

120
Particulate Matter (PM2.5)

24h

35

Table 6. 4: Malaysia, Canada and USA Ambient Air Quality Guidelines

Since the concentration of carbon dioxide emitted from our plant is

only 1.97 ppm, it is clearly below the discharge limit and thus its emission to
the natural environment will not bring any hazardous effect to living
organisms and also to the Mother Nature.
From the mass balance, it is clear that the effluent waste from
centrifuge 2 consists of biomass. Biomass is one of the most abundant
resources in this world. In this production, biomass consists of living or dead
cells of Cupriavidus necator H16, along with their metabolic constituents and
waste. Basically, biomass contains a lot of chemical energy within them,
where it can be converted to other form of energy, especially heat energy.
Since Cupriavidus necator H16 is known to be opportunistic bacteria and can
act as pathogens as well, our management planned to filter out all the
Cupriavidus necator H16 from the centrifuge 2 in negative pressure condition
and store them in a tightly enclosed storage tank in the biomass storage region.
Since biomass is renewable, we planned to sell them to fertilizer company,
where further treatment on the biomass can actually contribute to nutrient
production for the plants or in agricultural sector. Our target biomass buyer is
Sarawak Fertilizer Sdn. Bhd, a group of company incorporating at Kuching,
Sarawak. As per discussion, they agreed to pay RM 150 to RM 250 for per kg
of biomass since they will use the cells for further researches and for
transformation into fertilizers as well.
The third centrifuge and spray dryer generates sufficient amount of
wastewater composing of water, surfactant and hypochlorite. The wastewater
produced by these two equipment will be channeled to an activated carbon
filtration system placed in the wastewater treatment region of the plant. The
main purpose of channeling the wastewater mixture to the filtration system is
to filter out sodium hypochlorite, which has the same properties of bleaching
agent from the other two type of liquids. Then, the filtrate will be subjected to
further wastewater treatment mechanism. After some period of efficient

121
filtration, backwashing process will be conducted on the activated carbon in
order to flush out all the sodium hypochlorite that have been accumulated on
top of the filter (DeSilva, 2000). The solution containing sodium hypochlorite
is then sent to Kualiti Alam for further treatment.
The waste that can be send to Kualiti Alam has been categorized
differently by the management and the scheduled waste rates are classified
according to the type of waste. The information is listed as follows:

Table 6. 5: Characterization of Waste Type According to MIDA

These chemical wastes must be treated in accordance with the general

and specific standard. General prohibitions for discharging to sewers should
mainly be established to protect the collection systems, treatment works and
people as well as structures near the collection systems and treatment works,
banning the discharge of combustibles materials which may cause fire and

122
explosion; limiting grease and oils to prevent pipelines from being clogged;
prohibition of the discharge of hazardous substances to protect human health
and the environment. The effluent of the plant must reach the Department of
Environment specification, before it is can discharge to river or to atmosphere.

6.2

JABATAN ALAM SEKITAR (JAS) Schedule B and EQA

ENVIRONMETAL QUALITY ACT, 1974

Environmental Quality Act or EQA (1974) is the legislation related to
the prevention, abatement, pollution control and enrichment and enhancement
of the Malaysian environment. This act is mainly about the restriction of the
waste discharge into the environment in contravention of the acceptable
conditions.

6.2.1

Gaseous Emission
Under EQA (1974), industries that are in potential of emitting gaseous

or air pollutants, they are required to ensure that the emission of their gaseous
by-products to comply and obey with the following air emission standards for
the control of air pollution and gaseous emissions. The standards are:
i.

Stack Gas Emission Standards from Environmental Quality

(Clean Air) regulations 1978 ;

ii.

Recommended Malaysian Air Quality Guidelines (Ambient

Standards).

123

124

125

Under this aspect, our company did comply with all the standards,
where the emission of carbon dioxide gas from our plant is below the
discharge limit and thus no further treatment is needed as it is safe in nature.

6.2.2

Sewage, Industrial Effluent and Leachate Discharge

Under the regulations set in Environmental Quality Act (1974),

industries discharging sewage, industrial effluent and leachate are compulsory

to obey and comply with the following discharge limits:
i.

Sewage Discharge Standards

ii.

Industrial Effluent discharge limits

iii.

Leachate discharge standards

The standards are attached below according to the sequence of listing

done.

126

127

Appendix K2 is referred in the effluent management from our PHB

production plant, where through planned and organized waste treatment
system employed, all the discharge of waste effluents will be below the
discharge limits set for Standard B. Thus, the regulations of EQA is fully
obeyed in our company in order to ensure sustainability in economic growth
and nature wealth. All the measures taken before discharging effluent is
strictly done based on Environmental Regulations. Since PHB is an
environmental- friendly substance produced in our plant, we will never ever
sacrifice the harmonic relationship between nature and living organisms, just
for the sake of economic wealth at any means.

128
6.3

Waste Treatment Option

There are various ways to treat liquid or gaseous effluent in this PHB

production plant. The common and widely used treatment process in the
industry includes a wide range of choice, either biological, chemical or
physical methods. These methods have general criteria and will be further
explained in details.

6.3.1

Biological Method
Biological treatment process is widely adapted by plants that are

producing heavy stream of effluent with highly contaminant of soluble organic

impurities or a mix of the two types of wastewater sources. This treatment can
be divided into two major categories, aerobic and anaerobic process. Table 6.6
shows the comparison between these two types of process.
Table 6. 6: Comparison of Aerobic and Anaerobic Treatment (Mittal,
2011)
Parameter

Aerobic

Anaerobic

Microbial reactions take

Microbial reactions take

place in the presence of

place in the absence of

molecular/ free oxygen

molecular/ free oxygen

Reactions products are

Reactions products are

carbon dioxide, water

carbon dioxide, methane

and excess biomass

and excess biomass

Wastewater with low to

Wastewater with

medium organic

medium to high organic

impurities (COD < 1000

impurities (COD > 1000

ppm) and for

ppm) and easily

wastewater that are

biodegradable

difficult to biodegrade

wastewater e.g. food

e.g. municipal sewage,

and beverage

refinery wastewater etc.

wastewater rich in

Process Principle

Applications

129
starch/sugar/alcohol
Reaction kinetic

Relatively fast

Relatively slow
Relatively low

Net Sludge Yield

Relatively high

(generally one fifth to

one tenth of aerobic
treatment processes)

Typically direct
Post Treatment

discharge or
filtration/disinfection

Capital Investment

Example Technologies

6.3.2

Relatively high

Invariably followed by
aerobic treatment
Relatively low with pay
back

Activated Sludge e.g.

Continuously stirred

Extended Aeration,

tank reactor/digester,

Oxidation Ditch, MBR,

Upflow Anaerobic

Fixed Film Processes

sludge Blanket (UASB),

e.g. Trickling

Ultra High Rate

Filter/Biotower, BAF,

Fluidized Bed reactors

MBBR or Hybrid

e.g. EGSBTM, ICTM

Processes e.g. IFAS

etc.

Chemical Method
Generally, chemical treatment is according to the principle of oxidizing

the organic compound in the effluent using oxidant chemical to produce

fundamental product of oxidation such as carbon dioxide and water. Common
types of chemical treatment process are wet air oxidation, supercritical water
oxidation, ozonolysis and chlorinolysis. These methods are only efficient for
specific process especially inorganic contaminant. However, the usage of
chemical might produces unwanted and dangerous product if the wastewater
contains certain unpredicted compound that may react with the chemical used.
Although this rarely happens, but the possibilities is there and any assurance of
this will cause a very big problem. Economic wise, these chemicals less cost
efficient compared to the physical method.

130

6.3.3

Physical Method
Typical types of physical treatment process includes distillation,

evaporation, steam stripping, air stripping, liquid extraction, carbon adsorption

and resin adsorption. The main advantages of physical process in general are
its economic advantage, both in terms of capital investment and operating
costs, and simplicity of its design and low maintenance. However, the cons is
it is not very effective for the treatment of highly contaminated wastewater.
Furthermore, most industrial processes tend to produce high volume of
residual, which had to be treated by incineration, and this will cause extra cost.
However, incineration can be used for direct ultimate disposal of the waste on
its own individual operation without other pre-treatment physical instruments.

6.3.4

Selection of Method
Based on the criteria laid out of the three methods of treatment process,

it is very clear that the biological processes will be very suitable treatment for
this PHB production plant. For liquid waste treatment, activated sludge system
is chosen. Although it cannot be assured to be cost efficient, it is very effective
for organic waste treatment whilst the waste discharge will be more
environmental friendly.

131
6.4

Process Description

Figure 6. 2: Conceptual Flow Diagram for Activated Sludge Wastewater

Treatment System

The process implemented to manage waste from the plant is biological

treatment with activated sludge system. From Figure 6.2, the activated sludge
system consists of a primary clarifier, an aeration tank, a secondary clarifier,
and a disinfection region.
In this case, the wastewater from streams S-120, S-129 and S-134 will
be introduced to preliminary treatment and further fed to primary clarifier.
Primary clarifier functions to remove sediment solids from the wastewater.
The wastewater is then pumped into aeration tank. The aeration tank plays the
most important role for the treatment, with the presence of sufficient aeration,
the activated sludge or microorganisms will oxidize organic matter, both
soluble and particulate efficiently in a semi-controlled environment.
Subsequently, the treated wastewater will be channeled to the secondary
clarifiers. Here, the microorganisms and suspended particles are given enough
time to settle down. Excellent removal of organic matter is possible only by
proper designing and operation of second clarifier. It plays three major roles
such as thickening of biological solids for recycle, clarification of effluent
wastewater and also to store the biological mass in the settling tank. By fully

132
maximizing this component, particulate fraction that contributes for eventually
high BOD value of wastewater can be treated. Solid waste from primary and
secondary clarifiers will be sent to Kualiti Alam for disposal. Liquid waste
from secondary clarifier will be recycled to aeration tank. Finally, the treated
wastewater will be disinfected with addition of chlorine dosage and will be
nearly pure water and is safe to discharge into drainage system.

6.5

Waste in Polyhydroxybutyrate (PHB) Plant

The environmental impact due to day-to-day operation and the

potential environmental damage that would result from a plant accident or spill
have been considered. Potential emissions to the environment from the
proposed hepatitis B vaccine plant have been assessed in two categories,
airborne emissions and waterborne emissions. As we know the main product
of this plant is polyhydroxybutyrate (PHB), beside this product there are
effluent gaseous stream by-products streams. The effluent gaseous in this plant
is oxygen, carbon dioxide, and nitrogen. These effluents gaseous are treated in
a wet scrubber. The non-dangerous gaseous are released to the atmosphere.
The total amount of money used for waste management is still considerable in
our plant. Besides, we do generate revenue of RM 631,867 per year through
the selling of biomass.
For biomass, cell debris, and unwanted protein waste are sterilized first
in an autoclave to kill bacteria and impurities before send to Kualiti Alam for
waste management. Some of these products have commercial values, we plan
to sell these products. Selling these by-products will add the revenue of the
plant. We also have decided to send to Kualiti Alam for water and Water for
Injection waste for water treatment before they are discharged to environment.
All the wastes above cannot simply discharged to environment because it have
contaminants that can cause the pollution to environment.
Activated carbon filtration system will be purchased directly from the
supplier, Zhucheng Innovation Huayi, China. The cost of purchasing is
approximately RM 6279.48, whereas, the electricity and maintenance cost

133
annually amounts for RM 25,000.00. For the activated sludge wastewater
treatment system, the designs of each component and costing is done
according to W.M. Zahid (2007). The calculation for the cost of Kualiti Alam
Service is shown in Appendix E.

Table 6. 7: Total Gaseous Waste

WASTE

Carbon dioxide

TYPE OF

AMOUNT

WASTE

WASTE

(kg/year)

MANAGEMENT

Gas

4157.92

Wet scrubber

Gas

423,763

Wet scrubber

Gas

109,695

Wet scrubber

(0.80ppm)
Nitrogen
(81.7ppm)
Oxygen
(21.15ppm)
TOTAL

4581.683 kg/year

Table 6. 8: Total Waste Summary

WASTE

TYPE OF

AMOUNT

WASTE

(kg/year)

Biomass

Solid waste

45973.45

Autoclave Sarawak Fertilizer

Cell debris

Solid waste

81851.45

Autoclave Sarawak Fertilizer

TOTAL

WASTE MANAGEMENT

127824.89 = RM31,956.22 (Sarawak

Fertilizer: RM250/tonne)

Sodium

Chemical

189,076.25

Kualiti Alam / Sell to chemical

company

hypochlorite
water
TOTAL

Chemical

119,727.4
308,803

Kualiti Alam
= RM611,431 (Kualiti

Alam:RM1980/tonne)

134
Table 6. 9: Costing for Waste Treatment Option Employed in Our
Company
Treatment option/ Equipment

Cost (RM)

Activated Carbon Filtration System

6279.48

Electricity + Maintenance (per year)

25,000.00

Activated Sludge System

450,986.00 (W.M. Zahid, 2007)

Kualiti Alam service

30.96

Transportation service (Sg.Bako to

1500

Kuching )
TOTAL

483,796

135
CHAPTER 7
SITE SELECTION AND PLANT LAYOUT

7.1

Introduction
One of the crucial stages in the designing of plant is the selection of a

suitable site to create the secure and well developed plant as well as the
selection will result in a process in which the costs are minimized. Serious
consideration must be highlighted before locating a plant in Malaysia. The
site selection is to ensure adequate protection of site personnel, the public and
the environment from the impacts of the construction and operation of the
plant. Therefore, there are three locations that were identified and chose to
setup the polyhydroxybutyrate (PHB) plant which are:
i. Tanjung Langsat, Johor
ii. Sungai Bako area, Kuching, Sarawak

7.2

General Consideration of Plant Location

The final choice of the site location should be made after reviewing the

entire locations factor. There are several factors to be considered when

selecting a suitable site. Below are the lists of the factors to be considered:

Type of Industry Preferred and Location

Availability of Raw Material

Utility

Land Selling Price and Area Still Available

Transportation System

Availability of Manpower

Research and Development Organization

Geography, Climate and Environment

Government Incentive

Effluent Disposal

136

7.3

Type of Industry Preferred and Location

In doing this research on the suitable location to build up the bioprocessing

plant, the preferred location is the place that can provide a fully facilities for
running the plant process until the facilities provided for distribute it product
to the customers. It is important because all the facilities and incentives that
provide by federal government and state government are based on the types of
industries that can be run on that industrial area. Johor and Sarawak can be
suitable for bioprocessing plant as this type of the selected location is the
industrial area. So, by choosing these locations as a suggestion for setup a
PHB plant is the most suitable one.

7.3.1

Availability of Raw Material

Raw material is one of the most important factors in production. We

have to pay attention to the distance of the sources of raw material, the purity
of the raw material and the storage system that is applicable and suitable with
the characteristics of the raw material. In running the plant to produce PHB, it
is only one major raw material is used which is Jatropha oil. In Malaysia, there
are many Jatropha oil suppliers. But some of the large scale suppliers are
located in Sarawak, Selangor and Johor which are stated below:
Frozen Food Bhd

Frozen Food Bhd is established in 2005. It is based in Kuching,

Sarawak. It was leading exporter and supplier of high quality Whole
Frozen Chicken Halal from CIS with trademark. Other than Jatropha
oil, they too export worldwide the beef tallow, mutton tallow, frozen
chicken , chicken feet, whole chicken , buffalo beef , cattle beef, sugar,
, yellow and white corn, onion, sunflower oil, soybean cooking oil,
vegetable cooking oil, seeds and nuts and frozen food and meat
product.

Ganda Edible Oils Sdn Bhd

137

Ganda Edible Oils Sdn Bhd, is a manufacturer of edible oils such as

Palm oil, Sunflower oil, Soyabean Oil, Canola and also Jatropha oil for
biodiesel. It was established in 2007, and situated in Taman Mount
Austin, Johor Bahru.

Omega International Sdn Bhd Malaysia

Omega International SDN BHD Malaysia located at Muara Tabuan

Kuching, Sarawak. The company is a leading exporting trading
company in Malaysia. They deal directly with manufacturing company
that offers competitive prices. Other products than jatropha oil are
paint and coating chemicals, charcoal, dairy product and other office
and school suppliers.

7.3.2

Utilities
Utility cover the facilities of water supply and electricity supply. These

entire two components are important factors that ensure the smooth operation
of a plant. Most of the bioprocessing plant requires large quantities of water
for cooling and general process usage. Therefore the availability of water
reservoirs nearby must be ensured. Besides, the plant also needs electricity to
operate.

7.3.3

Water Supply
The location of our plant should be near with water supply with

minimum water shortage problem. We also have to consider the water quality
parameter such as temperature, mineral content and the probability of in need
of water treatment.
Table 7. 1: Water Provider Based on Location
Location

Provider

Supply capacity

Tanjung Langsat,

Sg Layang

180 MLD

Johor

Water Treatment

138
Sungai Bako area,

Batu Kitang Treatment

Kuching, Sarawak

Plant

7.3.4

166 MLD

Electricity Supply
Electrical power will be needed at all sites. The plant must be located

close to a cheap source of power. A competitively priced fuel must be

available on site for steam and power generation.
Table 7. 2: Electricity Provider Based On Location
Location

Tanjung Langsat, Johor

Provider

Supply capacity

Sultan Iskandar

630MW

400MW

210MW

114MW

Power Station
(TNB)

YTL Pasir
Gudang

Sungai

Bako

area,

Kuching, Sarawak

Sejingkat Power
Station, Kuching

Tun Abdul
Rahman Power
Station, Kuching

7.3.5

Land Selling Price and Area Still Available

The cost of the land depends on the location and varies between rural

and industrial area. We have to consider factors that make our plant strategic
towards the company production. We also need to consider the price of the
land while keeping in track with our company budget. The land should ideally
be flat, well drained and have suitable load-bearing characteristics.
Furthermore, the chosen site location should provide storage, and handling
infrastructure, emergency and other facilities as well.

139
Table 7.7: Land Prices Based On Location
Location

Land Selling Price

Land Size

Tanjung Langsat, Johor

RM 35,000

5 acres

RM 450,000

17.7 acres

Sungai Bako area,

Kuching, Sarawak

7.3.6

Transportation System
The transport of materials and products to and from the plant will be an

overriding consideration in site selection. If practicable, a site should be

selected that is close to at least two major forms of transport.
Table7.8: Transportation system based on location
Location
Tanjung Langsat, Johor

Facilities
Ports:

Located 5 km from Johor Port

Airport:

Around 57 km from Senai

International Airport

Road Network:

Around 35 km from raw

material supplier (Ganda
Edible Oils Sdn Bhd)

Sungai Bako area, Kuching, Sarawak

Ports:

Located near from Kuching

Port

Airport

Around 17 km to Kuching
International Airport

Road Network:

Around 20km from raw

140
material supplier (Frozen
Food Bhd)

7.3.7

Availability of Manpower
The manpower availability in the vicinity of the proposed site should

be considered. We required prevailing pay scales and suitable working hours.

Labour will be needed for construction of the plant and its operation.
Skilled construction workers will usually be brought in from outside the site
area, but there should be an adequate pool of unskilled labour available locally
and suitable labour for training to operate the plant. Skilled tradesman will be
needed for plant maintenance. Local trade union customs and restrictive
practices will have to be considered when assessing the availability and
suitability of the local labour for recruitment and training.

7.3.8

Research and Development Organization

From day to day, research and development (R&D) is important to

improve our production. By doing a research, a new thing could be recovered

and all this research will be conduct by any research centre or higher education
institution. It is important to have all this organization a round industrial area
to support the industrial in the future growth.

7.3.9

Geography, Climate and Environment

The types of land surface and adverse climatic conditions at a site will

increase cost. Abnormally low temperatures will require the provision of

additional insulation and special heating for equipment and pipe runs. Stronger
structures will be needed at locations subject to high winds or earthquakes.

141
7.3.10 Government Incentive
Capital grants, tax concessions and other inducements are often given
by the governments to direct new investment to preferred location. The
availability of such incentive can be the overriding consideration in site
selection.

7.3.11 Waste and Effluent Disposal Facilities

The plant should have satisfactory and efficient disposal system for
waste or industry effluent such as drainage system and the dumping site. Local
regulations will cover the disposal of toxic and harmful waste and
appropriated authorities must consult proper waste disposal system. Full
consideration must be given to the cost of the waste and effluent disposal.

7.4

Site Selection analysis

Based on the comparisons made in the Table 7.1, 7.2, 7.3 and 7.4, we

decided to choose land at Sungai Bako area Kuching, Sarawak as our future
plant site location. It is much bigger than the land at Tanjung Langsat, Johor.
Besides, Sungai Bako area Kuching, Sarawak located closer to our raw
material source. Moreover, there are many other suppliers in Sarawak
compared to in the peninsular Malaysia. This will help us to decrease the
transportation costs and total production costs and also saves time in term of
raw material delivery. The major Jatropha oil supplier will be from Omega
International SDN BHD Malaysia and Frozen Food Bhd. In addition, it also
has adequate supply of energy from Sejingkat Power Station, Kuching and
Tun Abdul Rahman Power Station, Kuching. While as for water supply comes
from Batu Kitang Treatment Plant. This will ensure smooth run of new plant.
The plant location chosen at Sungai Bako area Kuching, Sarawak is the most
strategic site for building new plant for production of PHB.

142
7.5

Plant Layout

7.5.1

Introduction
After deciding above the proper site for locating an industrial unit, next

important point to be considered is to decide about the appropriate layout for

the plant. Plant layout is primarily concerned with the internal set up of an
enterprise in a proper manner. It is concerned with the orderly and proper
arrangement and use of available resources viz., men, money, machines,
materials and methods of production inside the factory. A well designed plant
layout is concerned with maximum and effective 168tilization of available
resources at minimum operating costs. (Smriti Chand, 2014)
The term plant layout is used in broad sense to include factory layout
and machine layout. A plant layout refers to the arrangement of machinery,
equipments and other industrial facilities such as receiving and shipping
departments, tool rooms, maintenance rooms, employee amenities etc. for the
purpose of achieving the quickest and smoothest production of the lest cost.
(Himanshu. K.G, 2013).

7.5.2

Definition
Plant layout may be defined as physical arrangements of industrial

facilities. This arrangement includes the space needed for material movements,
storage, indirect labour and all other supporting activities or services as well as
operating equipment and personnel. Plant layout basically:
1. Placing the right equipment.
2. Coupled with the right method.
3. In the right place.
4. To permit the processing of a product unit in the most effective
manner, through the shortest possible distance, and in the shortest
possible time. (Himanshu. K.G, 2013)

143
7.5.3

Objectives of Plant Layout

The main objective consists of organizing equipment and working

areas in the most efficient way, and at the same time satisfactory and safe for
the personnel doing the work.
1. Sense of Unity
a. The feeling of being a unit pursuing the same objective.
b. Minimum Movement of people, material and resources.
2. Safety
a. In the movement of materials and personnel work flow.
3. Flexibility
a. In designing the plant layout taking into account the
changes over short and medium terms in the production
process and manufacturing volumes.
These main objectives are reached through the attainment of the
following facts:

Congestion reduction.

Elimination of unnecessary occupied areas.

Reduction of administrative and indirect work.

Improvement on control and supervision.

Better adjustment to changing conditions.

Better utilization of the workforce, equipment and services.

Reduction of material handling activities and stock in process.

Reduction on parts and quality risks.

Reduction on health risks and increase on workers safety.

Moral and workers satisfaction increase.

Reduction on delays and manufacturing time, as well as increase

in production capacity.

All these factors will not be reached simultaneously, so the best

solution will be a balance among them.

144
7.5.4

Factors Affecting the Plant Layout

a) Product and Material Specification: Material form, weight,

physical and chemical characteristics of material used has great influence in

locating a facility. These factors influence all parameters of the layout.

Products requiring hazardous and dangerous operations call for isolation of the
facility instead of integration on a line basis.
b) Location and site of the plant: Site location has big influence when
preparing layout. A plant has to get raw material from source and send the
products to the market. Hence the receiving department and the shipping
department must be properly located so that the receiving and shipping will
not become a problem. The attitude of the community, availability of power
supply and water has influence in locating the departments appropriately.
c) Manufacturing Process: The manufacturing process decide the
flow of material, types of machine required and the material handling facilities
required. Depending on the size and weight of the machine and raw material,
the working area required and additional space required to store in process
inventory has to be decided.
d) Material handling: Material handling need to keep as low as
possible to reduce the production cost. This point applies to all aspects of
manufacturing, for example, raw material handling, handling of in process
inventory, handling of scrap and waste and finished product. Material handling
and plant layout are closely related.
e) Storage on in-process inventory: Material waiting time should be
kept at minimum to eliminate unimportant cost. Layout should be designed in
such way that material does not wait in-between two processes.
f) Plant personnel and Employee facilities: Facilities like rest room,
vehicle parking facilities, caf and safety guards for machine should be
considered while preparing layout.

145
g) Work Areas and Equipment: Work areas are properly designed, so
that workers can move without much difficulty and material can be handled
with least difficulty. Sufficient place should be provide for maintenance
personnel to attend to the repairs or maintenance of the machines.
h) Disposal of waste and dangerous gasses: Proper waste disposal
facilities should be provided at appropriate palces, so that they are disposed to
a remote place, so that they wont be harmful to the workers or the community
living around the plant.
i) Flexibility: Factor of flexibility must be incorporate while preparing
the layout as time pass plant needs a chance. Change in product design or
change in material used or process lead to change in layout.( P. Rama Murthy,
2005)
The plant layout shall be arranged to:
1. Maximize safety;
2. Prevent spread fire
3. Facilitate easy operation and maintenance
4. Consider future expansion
5. Economize project
(KLM Technology Group, Feb 2011)

146

Figure 7. 1: Plant Layout of PHB plant

147
The most important factors of plant layout as far as safety aspects are
concerned are those to prevent, limit and/or mitigate escalation of adjacent
events (domino). Designed plant layout will ensure all the safety within on-site
occupied buildings by installation of amount of fire extinguisher. Control
access of unauthorized personnel can be done by security system which is
strictly applied. Emergency door enable facilitate access for emergency
services. Plant layout is designed fully according to all the factors and
objectives in order to reduce the production cost and ensure smooth operation
of whole plant. Table below show all the functions and location of designed
buildings and rooms in plant layout.
Table 7. 3: Building and Location in the Plant Layout
Buildings / Rooms/ Equipment
Offices

Office for management board

Manage the plant

Deal with supplier

Location

Located away from operation

plant for safety

Near the main entrance

Located at both main gate

Workshop

Near the plant area

Easy to conduct maintenance

Workshop

emergency on equipment;

malfunction, breakdown

Equally distribute most corner

Security room and post guard

Control and checking all

vehicle in and out

Record workers attendance

and visitors

Deliver pass for visitors

For mechanical and electrical

For maintenance purpose

Fire Assembly point

Fire assemble for place all

personnel and works placed if

area of plant

emergency happen
Control room

Manage and control all

Near plant building

148
machines and equipment in
operation line
Electrical Room

Main power control room

Enable maintenance of any

Located beside the processing

area

electrical supply issue

General Storage

For storage of sample of

product

Located beside laboratory

Near the processing area

Easy to transfer raw material

Storage of spare equipment or

apparatus need for production
line.

Raw Material Storage

To keep raw material in

suitable and safe condition

to process area
Laboratory

To analyze quality of the

product

Near processing area

Near processing area

Located beside the production

Do experiment for better

product

Chemical Storage

Storing chemicals that need

for production in safe and
suitable condition.

Waste treatment plant

Treating waste from

production line

area

Product Storage

Keep standard product in safe

and dry

Loading Area

Located near production area

149

Prepare transfer of product to

truck and deliver to customer

Located beside product

storage room

Generator Room

Temporary generate electric

power automatically to plant

Near Processing Area

Beside processing area

Near Processing area

when there is electric supply

disturbance
Biomass Waste Storage

Temporary store biomass

waste from production line in
safe condition

Packaging Room

Seal final quality product in

standard packaging

150
CHAPTER 8
ECONOMIC ANALYSIS

8.1

Introduction
Main objective of setting up a feasible plant is to gain profit. In this

chapter, it will focus on economic analysis where discounted cash flow is

evaluated. Economic potential were evaluated in previous chapters is to give
rough estimation of a plant profitability. Economic potential does not clearly
determine the actual economic performance as time value of money, plant
lifetime, or international and local trade regulations are not taken into
consideration. Economic analysis of this PHB plant has to be performed,
which is done by estimating fixed capital investment, total capital investment,
total production cost and revenue from sales.

8.2

Grass Root Capital

Grass root capital (GRC) is defined as the cost of equipment

installation in a plant. The term grass roots refers to a completely new facility
in which construction on essentially undeveloped land, a grass field. This
capital cost make up ajor portion of total fixed capital cost. In order to evaluate
the estimate cost of equipment involved in this PHB plant, bare module cost
(BMC) method is used. Table 8.1 summarizes the bare module cost for all
equipment and detailed calculations are shown in Appendix A
Table 8. 1: Bare Module Cost of Equipment in PHB Plant
Equipment

CBM (RM)

Gas Compressor G-101

34,762.34

Air Filter AF-101

12,475.32

Air Filter AF-102

12,475.32

Flow Splitter FSP-101

40,719.06

151
Flow Splitter FSP-102

1,105,663.95

Mixer MX-101

43,514.64

Mixer MX-102

72,742.71

Heat Sterilizer ST-101

12,475.32

Heat Sterilizer ST-102

12,475.32

Seed Fermenter V-101

72,194.35

Main Fermenter V-102

663,087.65

Storage V-103

55,018.36

Blending Tank V-104

12,360.99

Blending Tank V-105

91,835.10

Centrifuge DS-101

79,689.43

Centrifuge DS-102

88,198.73

Centrifuge DS-103

81,212.16

Pump PM-101

23,067.68

Pump PM-102

31,239.37

Pump PM-103

5,763.94

Pump PM-104

5,763.94

Pump PM-105

5,763.94

Pump PM-106

5,763.94

Spray Dryer SDR-101

17,804.03

Total Bare Module Cost, CTBM

2,586,067.58

The total module cost can be evaluated from (Turton et al., 2009)
CTM = 1.18(CTBM)

----- (8.1)

And the grass root costs can be evaluated from (Turton et. al., 2009)
CGR = CTM + Auxiliary Cost ----- (8.2)
In addition to direct and indirect costs, it is necessary to take into
account for other cost such as contingency and fee cost, and auxiliary facility
cost. Table 8.2 shows estimation of GRC.

152
Table 8. 2: Estimation of Grass Root Capital, GRC.
Investment

Cost (RM)

Contingency and Fees (8% of CTBM)

206,886.00

Auxiliary and Facility (10% of

279,295.00

CTBM)
Total Module Cost (CTM)

3,051,560.00

Grass Root Capitals (CGR)

3,356,716.00

8.3

Capital Investment
It is known that a lot of expenses involved in purchasing and installing

equipment and facilities before an industrial plant can fully operate. Land and
service facility must be obtained beforehand followed by installing equipment,
piping and control system, and so forth for smooth operation of plant.
Therefore, money is crucial in the build-up and operation of the plant.
For build-up plant, the capital involved in the expenses of
manufacturing nad plant facilities is called fixed capital investment (FCI). On
the other hand, the capital used for operation of the plant is known as the
working capital. The sum of these capitals is called total capital investment
(TCI).

8.3.1

Fixed and Total Capital Investment

FCI represents the capital that used for installed process equipment

with all auxiliaries that necessary for the process operation. It is categorized
into two categories; direct and indirect cost. FCI value is obtained by summing
the GRC with the direct and indirect costs. Direct cost is the cost that used for
purchasing and installing equipment with all piping system and control system
and also the expenses on land and service facilities; while indirect cost refers
to the cost pay to contractors and others. The formula to obtain TCI is as
shown in Equation 8.3

153
TCI = FCI + Working Capital + Start Up ----- (8.3)
Working Capital (WC) consists of the total amount of money invested
in raw materials and supplies carried in stock, finished products in stock and
semi-finished products in the process of being manufactured, account
receivable, cash kept on hand for monthly payment of operating expenses, e.g.
salaries, wages and raw material purchases. Table 8.3 shows the amount of
fixed and total capital investment of this PHB plant.
Table 8. 3: Fixed and Total Capital Investment
Total Capital Investment
Factor of GRC

Direct Cost
Onsite
Purchased Equipment Installation
Piping (installed)
Instrumentation and Control (installed)
Electrical and Material (installed)
Offsite
Building
Land
Service Facilities
Total

Indirect cost
Contingency
Construction Expenses
Engineering and supervision
Contractors Fee
Total
Total Cost = Direct Cost + Indirect Cost
Fix Capital Investment (FCI)
Working Capital
Total Capital Investment (TCI)

8.4

Cost (RM)

0.06
0.04
0.02
0.02

214,126.40
142,750.93
71,375.47
71,375.47

0.02
0.01
0.08

71,375.47
35,687.73
285,501.86
892,193.32

Factor of GRC
0.05
0.03
0.02
0.02

Cost (RM)
178,438.66
107,063.20
71,375.47
71,375.47
428,252.79
1,320,446.11
Total Cost + GRC 4,889,219.37
10% FCI
488,921.94
5,378,141.31

Manufacturing Cost
Manufacturing cost comprises of direct manufacturing expenses,

general manufacturing cost, manufacturing expense, annual depreciation, total

expenses, revenue from sales and net annual profit, as well as rate of return.

154
This manufacturing cost has to be paid by investors per year in order to
produce constant production of PHB. Direct manufacturing costs are costs
represent operating expenses that vary with production rate. It comprises of
cost of raw materials, waste treatment, utilities, operating labor, direct
supervisory and clerical labor, maintenance and repairs, laboratory charges,
and patents and royalties (Turton et al., 2009).
i.

Operating labor refers to people who actually run the equipment. It

estimates the amount of workers that are needed in the plant as well
as labor cost required.

ii.

Direct supervision and clerical labor usually include the cost of

administrative, engineering, and personnel support. It is 10% of
operating labor cost.

iii.

Maintenance and repairs constitute an important and necessary

budget item in the plant and assume to be costs 2% of fixed capital
investment.

iv.

Operating supplies include replaceable materials such as instrument

charts, lubricants, custodial supplies, and other items not
considered as part of regular maintenance. It is suggested that it
costs 0.5% of fixed capital investment.

v.

Laboratory expenses result from quality control testing and

chemical or physical analyses for product improvement. It is 10%
of operating labor cost.

vi.

Patents and royalties is the cost of using patented or licensed

technology which is 6% of total manufacturing cost.
According to Turton et al. (2009), fixed manufacturing cost is defined

as the cost that are independent of change in production. It comprises of plant

overhead cost, local taxes, insurances, and depreciation.
i.

Plant overhead cost is catch-all costs associated with operations

of auxiliary facilities supporting the manufacturing process.
The cost is summation of 70.8% operating labor cost and 3.6%
fixed capital cost.

155
ii.

Local taxes and insurance is the cost associated with property

taxes and liability insurance. It usually based on the plant
location and severity of the process. It could be 1.4% of fixed
capital investment.

iii.

The value of plant will depreciate year after year. Thus it is

consider depreciation as 10% of fixed capital investment.

In addition to direct and fixed manufacturing cost, a certain portion of

corporate management cost, sales expense, and research effort must be
financed from the plant. Among them are the expenses are administrative cost,
distribution and selling expenses, as well as research and development cost
(Turton et al., 2009).
i.

Administrative cost is the costs for administration procedures

which includes salaries, other administration, building and other
related activities. It assumed to be summation of 17.7% operating
labor cost and 0.9% fixed capital investment.

ii.

Distribution and selling expenses are the costs of sales and

marketing required to sell chemical products which is 11% of total
manufacturing costs.

iii.

Research and development costs are the costs of research activities

related to the process and products which include salaries and
funds for research related equipment and supplies. It could be 5%
of total manufacturing costs.
Manufacturing cost can be calculated by using formula in Equation 8.4

COM = 0.280FCI + 2.73COL + 1.23 (CUT + CWT + CRM) ----- (8.4)

Table 8.4 shows estimation of operating labor cost, while Table 8.5
summarizes the summary of manufacturing cost.
Table 8. 4: Estimation of Operating Labor Cost
Equipment Type

No. of Equipment

Compressor

Air filter

156
Sterilizers

Fermenters

Centrifuges

Blending Tanks

Spray Dryers

Total (Nnp)

13

Number of operators per shift, NOL can be calculated by using this formula;
NOL = (6.29 + 31.7P2 + 0.23Nnp)0.5
where
P: Number of particulate processing step
Nnp: Number of non-particulate processing step
Number of operators per shift is 22.726 or 23. For three shifts a day,
number of operators required per day is 68 and since average wages for
operators is RM850/month, total labour cost per year is about RM 695,421.61.
Table 8. 5: Summary of Manufacturing Cost
Manufacturing Cost Summary
RM/kg

Amount (kg/year)

350

50000

PHB

Sale Revenue
(RM/year)
17,500,000

Direct Manufacturing Cost

No. of batches per year =
Raw Materials

130
Amount, kg/batch

Amount,

RM/k

Total

kg/year

(RM/year)

Jatropha oil

390

50700

2.73

138,411.00

Urea

0.25

32.5

10.50

341.25

Total Cost of Raw Materials

138,752.25

Waste Treatment
Activated Carbon Filtration System

8,052.50

Electricity + Maintenance

20,000.00

157
Activated Sludge System

451,000.00

Kualiti Alam Service

2,500,000.00

Transportation

100,000.00

Total Waste Treatment Cost

3,079,052.50
Utilities

Power
Equipment

Consumption
(kW/batch)

Industry
tariff
(Tenaga

0.38

MYR/kWh

Nasional)

P-1/G-101

172.79

8000

hour/year

Pumps

522.26

130

btch/year

P-11/PV-105

10.58

23.38

P-14/PV-106

13.38

Total usage

719.01

Total usage per year

93,471.09

kW/year

Total Electricity Cost

16,813.73

MYR/year

MYR/kW/bat
ch

Industry
Equipment

Steam supply

tariff

(kg/h)

(Malaysian

0.62

MYR/kg

0.1

MYR/m3

1000

kg/m3

Gas)
P-3/V-101

43.49

P-8/ST-101

187.20

P-4/V-101

266.95

P-6/V-102

626.53

Total usage

1,124.17

Total usage per year

8,993,360.00

kg/year

Total Steam Cost

5,575,883.20

MYR/year
Industry

Equipment

Water supply
(kg/batch)

Tariff
(Lembaga
Air
Sarawak)

P-5/SFR-101

113.84

P-4/V-101

418.06

P-6/V-102

3,162.56

P-16/V-104

9,155.05

Total usage

12,849.51

Water
density

kg/batch

158
Total usage per year
Total Water Cost

1,670,436.44

kg/year

167.04

MYR/year

Total Utilities COST

5,592,863.98

FCI

4,889,219.37

COL

695,421.61

Cost of Manufacturing, COM

Maintenance and repairs

0.280FCI + 2.73COL + 1.23 (CUT + CWT +

14,104,604.9

CRM)

2% FCI

97,784.39

0.5% FCI

24,446.10

10% COL

69,542.16

Laboratory charges

10% x COL

69,542.16

Patents and Royalties

6% X COM

846,276.30

Operating supplies
Direct Supervision & Clerical
Labor

Total Direct Manufacturing Cost (TDMC)

10,496,073.75

Fixed Manufacturing Cost

0.708COL +

Plant Overhead Cost

668,370.40

0.036FCI

Local Taxes and Insurance

Depreciation

0.014FCI

68,449.07

0.1FCI

488,921.94

Total Fixed Manufacturing Cost, TFMC

1,313,747.35

General Manufacturing Cost

Administration Cost
Research & Development

0.177COL +

167,092.60

0.009FCI
0.05COM

705,230.25

Total General Manufacturing Cost, TGMC

2,423,829.39

Annual Manufacturing Expenses, AME

14,233,650.50

Annual Net Profit, ANP

Income Taxes

Revenue from sales - ATE

3,266,349.50

30% ANP

979,904.85

Net Annual Profit, ANNP

2,286,444.65

Rate of Return, I%
(ANNP + ABD) / TCI
100% =

52%

From Table 8.5, it is determined that the rate of return (ROR) is 52%.

159

8.5

Cash Flow Analysis

The final step in the profitability analysis is to determine the payback

period (PBP), discounted break event point (DBEP), and net present value
(NPV) of the PHB plant. Some assumptions are made in this cash flow
analysis:
i.

2 years start-up is required for a new PHB plant.

ii.

PHB plant operation life is 20 years.

iii.

30% of total capital investment (TCI) is expended on the plant first

start-up year and 70% of TCI is expended in the second start-up
year.

iv.

30% of the federal income tax.

v.

Sales income for the first year after start-up is 80% of the targeted
value.

8.5.1

Payback Period Analysis

Payback period (PBP) is defined as the length of time in which the

initial cash outflow of an investment is expected to be recovered from the cash

inflows generated by the investment. In terms of cash flow analysis, PBP is the
time required after start-up until cumulative undiscounted cash flow repays
fixed capital investment. PBP is important since it can determine the
feasibility of a PHB plant. Table 8.6 shows the cash flow analysis for
undiscounted rate (I=0%) and Figure 8.1 illustrates the cash flow diagram with
the payback period. Table 8.7 shows cash flow analysis for various discounted
rate and Figure 8.2 shows the discounted cash flow diagram according to FCI
is RM 4,889,219.37 and working capital at the end of year 2 is RM
488,921.94. In the Figure 8.1, PBP is determined as 4.5 years.

160

Undiscounted Cash Flow Analysis

Annual Capital
Year

Investment
A(I)

Sales Income

Depreciation

AS

AD

Total
Expanses
APC

Cash Income
AS-APC

Net Profit

Federal

(AS-APC)-

Income

AD

Taxes AIT

Net Profit
after Taxes

Net Cash

(AS-APC-

Income

AD)-AIT

Summation
Net Cash
Income

537,814.13

- 537,814.13

- 537,814.13

5,867,063.75

- 5,867,063.75

- 6,404,877.88

14,000,000.00

488,921.94

14,233,650.50

- 233,650.50

- 722,572.43

- 36,128.62

- 686,443.81

- 686,443.81

- 7,091,321.69

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

- 4,452,765.50

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

- 1,814,209.31

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

824,346.88

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

3,462,903.07

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

6,101,459.26

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

8,740,015.44

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

11,378,571.63

10

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

14,017,127.82

11

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

16,655,684.01

12

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

19,294,240.20

13

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

21,932,796.39

14

17,500,000.00

488,921.94

14,233,650.50

3,266,349.50

2,777,427.57

138,871.38

2,638,556.19

2,638,556.19

24,571,352.58

Table 8. 6: Cash Flow Analysis for Undiscounted Rate, I%

161

Undiscounted Cash Flow

30,000,000.00
25,000,000.00

Cumulative Cash Flow (RM)

20,000,000.00

Payback Period = 4.5 years

15,000,000.00
10,000,000.00
5,000,000.00
0

(5,000,000.00)
(10,000,000.00)

Year

Figure 8. 1: Undiscounted Cash Flow

10

11

12

13

14

15

162
Table 8. 7: Discounted Cash Flow Summary
Discounted Factor Cash flow Analysis for 10.00%, 20.00% and 30.00%
Discount
Year

Net Cash Income

Factor

Discounted Cash

Discounted Cash

Discount

Discounted

Discounted Cash

Discount

Discounted

Discounted Cash

Flow for 10%

Factor

Cash

Flow for 20%

Factor

Cash

Flow for 30%

Cumulative

fd (20%)

Flow for 20%

Cumulative

fd (30%)

Flow for 30%

Cumulative

448,178.44

0.77

- 413,703.18

413,703.18

Flow for 10%

fd (10%)
0

- 537,814.13

0.91

488,921.94

- 5,867,063.75

0.83

- 686,443.81

0.75

2,638,556.19

0.68

1,802,169.38

2,638,556.19

0.62

2,638,556.19

488,921.94

0.83

4,848,813.01

- 5,337,734.95

0.69

- 4,074,349.82

- 4,522,528.27

0.59

-3,471,635.35

3,885,338.53

515,735.40

- 5,853,470.35

0.58

- 4,919,775.84

0.46

- 312,445.98

4,197,784.51

- 4,051,300.97

0.48

1,272,451.87

- 3,647,323.98

0.35

923,831.86

3,273,952.64

1,638,335.80

- 2,412,965.17

0.40

1,060,376.55

- 2,586,947.42

0.27

710,639.90

2,563,312.75

0.56

1,489,396.18

0.33

883,647.13

- 1,703,300.29

0.21

546,646.07

2,016,666.67

2,638,556.19

0.51

1,353,996.53

430,427.54

0.28

736,372.61

966,927.69

0.16

420,496.98

1,596,169.69

2,638,556.19

0.47

1,230,905.94

1,661,333.48

0.23

613,643.84

353,283.85

0.12

323,459.22

1,272,710.48

2,638,556.19

0.42

1,119,005.40

2,780,338.87

0.19

511,369.87

158,086.02

0.09

248,814.78

1,023,895.70

2,638,556.19

0.39

1,017,277.63

3,797,616.51

0.16

426,141.56

584,227.58

0.07

191,395.99

832,499.71

10

2,638,556.19

0.35

924,797.85

4,722,414.35

0.13

355,117.96

939,345.54

0.06

147,227.68

685,272.03

11

2,638,556.19

0.32

840,725.32

5,563,139.67

0.11

295,931.64

1,235,277.17

0.04

113,252.06

572,019.97

12

2,638,556.19

0.29

764,295.74

6,327,435.41

0.09

246,609.70

1,481,886.87

0.03

87,116.97

484,903.00

13

2,638,556.19

0.26

694,814.31

7,022,249.72

0.08

205,508.08

1,687,394.95

0.03

67,013.05

417,889.94

14

2,638,556.19

0.24

631,649.37

7,653,899.10

0.06

171,256.73

1,858,651.68

0.02

51,548.50

366,341.44

923,568.99

448,178.44

397,247.58

163

Discounted Cash Flow

30,000,000.00

Cumulative Cash Flow, RM

25,000,000.00
20,000,000.00
15,000,000.00

10,000,000.00
5,000,000.00
0

(5,000,000.00)
(10,000,000.00)

Year
0%

10%

20%

30%

Figure 8. 2: Discounted Cash Flow

10

11

12

13

14

164
Table 8. 8: Net Present Value for Discounted Rate
Interest, I (%)

NPV (RM)

24,571,352.58

10

7,653,899.10

20

1,858,651.68

30

366,341.44

Discounted break-even point is when the time that used for discounted
cumulative cash flow become positive after decided to proceed which the total
income is higher than total production cost (Ulrich, 1984). Net present value is
the final cumulative of the annual discounted cash flow value at project
conclusion (Saravacos & Kostaropoulos, 2002). When the fractional interest
rate for which net present value equals to zero after number of years, the
discounted break-even point is called discounted cash flow rate of return
(DCFRR). The DCFRR also knows as the profitability index, initial rate of
return or investors rate of return (Saravacos & Kostaropoulos, 2002). This
rate of return is equivalent to the maximum interest rate (which is normally,
after taxes). This is because money could be borrowed to finance the project
under conditions where the net cash flow to the project over its life would be
just sufficient to pay all principal and interest accumulated on the outstanding
principal (Peters & Timmerhaus, 2004). Then, the graph of cumulative
discounted annual cash flow at different rate of return versus time is plotted
and shown in Figure 8-2. The discounted cash flow rate of return is
interpolated from values in Table 8.8 and it is obtained that 28.35%. Based on
the Figure 8.1 and the calculation, the payback period for this PHB plant is 4.5
years.

165

Discounted Factor Cash flow Analysis for 28.35%

Year

Net Cash Income

Discount Factor

Discounted Cash

Discounted Cash

fd (28.35%)

Flow for 28.35%

Flow for 28.35%

Cumulative

537,814.13

0.77912

419,021.53

419,021.53

5,867,063.75

0.60703

3,561,468.12

3,980,489.65

686,443.81

0.47295

324,651.47

4,305,141.12

2,638,556.19

0.36848

972,260.98

3,332,880.14

2,638,556.19

0.28709

757,507.58

2,575,372.56

2,638,556.19

0.22368

590,189.00

1,985,183.56

2,638,556.19

0.17427

459,827.81

1,525,355.75

2,638,556.19

0.13578

358,260.86

1,167,094.89

2,638,556.19

0.10579

279,128.06

887,966.84

2,638,556.19

0.08242

217,474.14

670,492.70

10

2,638,556.19

0.06422

169,438.36

501,054.34

11

2,638,556.19

0.05003

132,012.75

369,041.59

12

2,638,556.19

0.03898

102,853.72

266,187.87

13

2,638,556.19

0.03037

80,135.35

186,052.52

14

2,638,556.19

0.02366

62,435.02

123,617.50

Table 8. 9: Discounted Cash Flow at DCFRR=28.35%

166

167

Discounted Cash Flow

25,000,000.00

Cumulative Cash Flow, RM

20,000,000.00
15,000,000.00

10,000,000.00
5,000,000.00
-

(5,000,000.00)
(10,000,000.00)

Year

0%

10%

20%

30%

10

11

12

13

14

165
8.6

Profitability Analysis
A sustainable business and mission requires effective planning and

financial management. Ratio analysis is a useful management tool that

provides financial results and trend over time to a company. It also acts as a
key indicator of organizational performance. From this analysis, engineers
able to pinpoint the strengths and weaknesses from which the strategies and
initiatives can be formed. Profitability ratio includes gross profit margin,
operating profit margin and net profit margin.
Cost of goods sold is the costs that attribute to the products sales by a
company or industry, therefore the only costs included in the measure are
those that directly tied to the production of products. This cost including cost
of material, labor, plant overhead and other manufacturing costs but excludes
indirect expenses such as distribution and sales force costs.
The operating margin is the ratio of operating profit (operating income,
income before interest and taxes) to sales. This is a ratio that indicates how
much of each dollar of sales is left over after operating expenses. The
operating profit margin gives the business owner a lot of important
information about the firms profitability, particularly with regard to cost
control. A high operating profit margin means that the company has good cost
control and that sales are increasing faster than costs, which is the optimal
situation for the company.
The profit margin can measure how much profit a company makes for
every $1 it generate in revenue or sales. Profit margins vary by industry, but
all else being equal, the higher companys profit compare to its competition,
the better. The net profit margin is the ratio of net income to sales, and
indicates how much of each dollar of sales is left over after all expenses.
Gross Profit Margin
Cost of Goods Sold
10,732,161.25
Sales
17,500,000.00
The gross profit margin is:
38.67%
Operating Profit Margin
Operating profit
9,641,778.64
Sales
17,500,000.00

166
The operating profit margin is:
44.90%
Net Profit Margin
Net Profits After Taxes
13,349,245.05
Sales
17,500,000.00
The net profit margin is:
23.72%
8.7

Conclusion
The Total Capital Investment (TCI) for this PHB plant is RM

5,378,141.31 and the Total Expenses (ATE) is RM 14,233,650.50. The net

annual profit after taxes (ANNP) is RM 2,286,444.65. The plant spends 2
years for start-up and its operating life is 15 years with a depreciation of 10%
of fixed capital investment. For undiscounted cash flow, the payback period is
4.5 years. The rate of return obtained after the taxes is 28.35%.

167

CHAPTER 9
CONCLUSIONS AND RECOMMENDATION

9.1

Conclusion
The production of 50 MT per annum of Polyhydroxybutyrate has a

bright future. The PHB demand will be increased to 1025 metric tons in 2015
with 27.9% of annual growth. Due to the unique features, production of large
scale of PHB gained a lot of interest globally, with the increasing
environmental awareness among all the nations worldwide. An easy to handle
and low cost production process is fundamental if a successful
commercialization similar to plastics is intended.
For the process synthesis and flow sheeting, base case material and
energy balance has been performed by manual calculation. Simulation was ran
by using Superpro software and the result was compared with manual
calculation. Detailed equipment sizing and design of all major equipment also
had been performed using fundamental chemical engineering principles. A
highly integrated process control system was also included to the proposed
plant.
In respond to the environmental responsibility, the plant has been
designed to achieve the target of waste minimization and cost minimization.
The unwanted side product is being treated to ensure the emission coming out
from the plant has met the standard of environmental act.
On the safety aspect, hazard and operability (HAZOP) study has been
performed to identify on operability problems and providing necessary
resolution. A general safety study includes personal safety, emergency
management and plant start-up and shut down procedures.
The total capital cost needed is RM 5,378,141.31. The manufacturing
cost comprises RM 14,233,650.50. With the current market value for high
purity PHB of RM 35.00/tons, the net revenue has been calculated to attain

168
RM 17,500,000.0, thus it is obvious that the production of 50 metric tons of
PHB is feasible enough to be established. Based on financial analysis done, we
are able to attain 28.35% rate of return with the payback period of 4.5 years.
Finally, it can be concluded that the construction of a 50 MT/year PHB
production plant in the area of Sungai Bako, Kuching, Sarawak is technically
feasible and economically attractive. This plant will get good support and
encouragement from the government.

9.2

Recommendation
As mentioned in previous chapters, complete study had been done in

all aspects in determining the feasibility of establishing a 50 MTPA PHB

production plant in Malaysia. However, there is always room for improvement
in order to aid in the betterment of this plants effective operation. Firstly,
safety considerations can be fully upgraded. More evaluations and thorough
study on HAZOP can be implemented continuously. Fault-Tree Analysis
(FTA) can also be applied to this aspect, to ensure the safety features of all the
production and separation units in this plant can be improved. Besides, it can
also ensure higher safety assurance to the labors.
Secondly, more effective and deeper studies on finding new
alternatives either in the supply of nutrient or in the downstream processing of
PHB recommended to be carried out. This matter is essential so that the
production of PHB by Cupriavidus necator can be increased more but with
lower production cost. Significant development work can be implemented in
the PHB production using non-edible plant oil such as Jatropha oil. The
combined properties of few biodegradable plastics can turn out to be efficient
in many ways.
Lastly, waste management must be always studied and improved from
time to time. The application of activated sludge wastewater treatment system,
although known to be effective in providing higher quality of treated water in
biological

process,

yet

it

is

not

cost-effective

method.

Thus,

169
phytoremediation technology, which is proven to be easy, inexpensive and
effective can be implemented. All the measures for pollution control must be
always renewed, so that there is no room for causing any harm to environment.

vi

REFERENCES
Abe, H.,Doi, Y (2002), Molecular and material design of biodegradable
polyhydroxylalkonate

Akiyama M, Tsuge Y, Doi Y. Environmental life cycle comparison of

polyhydroxyalkanoates produced from renewable carbon resources by
bacterial
fermentation. Polymer Degradation & Stability
2003; 80:183-94

A.P. Bonartsev, V. M. (2007). Biosynthesis, biodegradation, and application

of poly(3-. Communicating Current Research and Educational Topics
and Trends in Applied Microbiology, 295-307.

A.Ugur,N.Sahin,Y.Beyatli (2001),Accumulation of poly-hydroxybutyrate in

Streptomyces species during growth with different nitrogen
sources,26,171.

Batcha AFM, Prasad DMR, Khan MR, Abdullah H. Biosynthesis of poly(3hydroxybutyrate)(PHB) by Cupriavidus necator H16 from jatropha oil
as carbon
source. Bioprocess Biosyst Eng 2013; DOI
10.1007/s00449-03-1066-4

BIONAS. (2014, April 25). BIONAS. Retrieved April 25, 2014, from BATC
Development Berhad
: http://bionas- usa.com/file/BATC%20%20Company%20Profile.pdf

Bioplastics, E. (2014, April 24). European Bioplastics Corporation. Retrieved

April 24,
from European Bioplastics Web site: http://en.europeanbioplastics.org/market/europebeyond/

vii
Christopher J. Brigham, a. A. (2012). Applications of Polyhydroxyalkanoates
in the Medical Industry. International Journal of Biotechnology for
Wellness Industries, 56-58.

Choi JI, Sang YL. Process analysis and economic evaluation for Poly(3hydroxybutyrate)
production by
fermentation.
Bioprocess
Engineering 1997;17:335-342

Daniel

A.
Crowl,
Joseph
F.
Louvar,
(2011),
Chemical Process Safety: Fundamentals with Applications. Hall
International Series in the Physical and
Chemical Engineering Sciences
(3).

Development, O. f.-o. (2013). Working Party on Biotechnology . POLICIES

FOR BIOPLASTICS IN THE CONTEXT OF A BIOECONOMY ,
17.

D.S. Rosa, N.T. Lotto, D.R. Lopes, C.G.F. Guedes(2004), The use of
roughness for evaluating of poly-b-(hydroxybutyrate) and poly-b(hydroxybutyrate-co-b-valerate),
Polym. Test. 23 .

Du G, Chen J, Yu J, Lun S. Continuous production of poly-3-hydroxybutyrate

by Ralstonia eutropha in a two-stage culture system. Journal of
Biotechnoogy 2001;88:59-65

Fitzgerald M. Indias big plans for biodiesel. Available from:. Technology

Review:
Massachusetts
Institute
of
Technology
http://www.technologyreview.com/Energy/17940/page1/; 2006.

Fraser, D. M., & Hallale, N. (2000). Determination of Effluent Reduction and

Capital Cost Targets through Pinch Technology. Environmental
Science & Technology, 34(19),
41464151.

G.D. Saravacos and A.E. Kostaropoulos, (2002) Handbook of Food

Processing
Equipment , Kluwer Academic/Plenum Publishers

viii

Gironi F, Piemonte V. Bioplastics disposal: how to manage it,Tallinn; 2010.

Gui MM, Lee KT, Bhatia S. feasibility of edible oil vs, non-edible oil
vs. waste edible oil as biodiesel feedstock. Energy 2008;33:379-82

Himanshu. K.G,( 2013),Facilities layout (plant layout), Retrieved on 3 May

from:
http://blogs.siliconindia.com/himanshugoelmba/plant_layoutbid-6stv5c2H28957491.html

Hoh, R. (2012). Oilseeds and product annual. Global Agricultural Information

Network.
USDA Foreign Agricultural Service.

Jong-il Choi, S. Y. (1977). Process analysis and economic evaluation for

Poly(3-hydroxybutyrate) production by fermentation. Bioprocess
Engineering, 335-342.

Jim C. Philp, Alexandre Bartsev1, Rachael J. Ritchie2, Marie-Ange Baucher

and K. Guy. Bioplastics science from a policy vantage point
2013;30: 1-2

Kadouri D, Edouard J, Okon Y. Ecological and agricultural significance of

bacterial
polyhydrxyalkanoates. Critical Review Microbial
2005;31:55-67

Kahar P, Tsuge T, Taguchi K, Doi Y. High yield production of

polyhydroxyalkanoates from soybean oil by Ralstonia eutropha and its
recombinant strain. Polymer Degradation & Stability 2004;83:79-86

Kahar, P., Tsuge, T., Taguchi, K., Doi, Y. (2004). High yield production of
polyhydroxyalkanoates from soybean oil by Ralstonia eutropha and its
recombinant strain. Polymer Degradation and Stability. 83(1): 79-86

Kek, Y.K., Chang, C.W., Amirul, A.A., Sudesh, K., (2010) Heterologous
expression of Cupriavidus sp. USMAA2-4 PHA synthase gene in
PHB4 mutant for the production of poly(3-hydroxybutyrate) and

ix
its copolymers. World Journal of Microbiology and Biotechnology.
26(9):1595-1603

Kek YK, Lee WH, Sudesh K. Efficient bioconversion of palm acid oil and
palm kernel oil to
poly(3-hydroxybutyrate)
by
Cupriavidus
necator. Can J Chem 2008;86:533-9

KLM Technology Group,(Feb 2011), Layout Spacing (Project Standard And

Specification),
Project Engineering Standard

Khan MR, Prasad DMR, Abdullah H, Batcha AFM. Kinetic analysis on cell
growth and biosynthesis of poly(3-hydroxybutyrate)(PHB) in
Cupriavidus necator H16.
International Journal of Bioscience,
Biochemistry and Bioinformatics 2013;3:516518

Khanna S, Srivastana AK. Statistical media optimization studies for growth

and PHB
production
by Ralstonia eutropha. Process Biochem
2005;40:2173-82

Ko-Sin Ng, W.-Y. O.-K. (2010). Evaluation of jatropha oil to produce poly(3hydroxybutyrate). Polymer Degradation and Stability, 1.

Lee WH, Loo CY, Nomura CT, Sudesh K. 2008. Biosynthesis of

polyhydroxyalkanoate
copolymers from mictures of plant
oils and 3-hydroxyvalerate precursors.
Bioresource Technology
;99:44-51

Lemoigne, M. Produits de dehydration et de polymerisation de lacide oxobutyrique. Bull Soc Chim Biol, 1926, 8, 770782.

Li Hongqi, Shen, Park et al. (1999). Production of Poly(3-Hydroxybutyrate)

by Fed-Batch Culture of AlcaligenesEutropus at Phosphorus
Limitation in 50L fermenter. Chinese
J. of Chem. Eng., 7(4)
368-371

x
Loo CY, Lee WH, Tsuge T, Doi Y, Sudesh K.( 2005)Biosynthesis and
characterization of
po;y(3-hydroxybutyrate-co-3hydroxyhexanoate) from palm oil products in a
Wautersia
eutropha mutant. Biotechnol Lett;27:1405-10

Malaysian Palm Oil Board, MPOB (2008). World Production Malaysian Palm
Oil Board,
MPOB
(2008). World production of 17 oils &
fats: 1999 2008. Available online
at
http://econ.mpob.gov.my/economy/annual/stat2008/World6.3.pdf
[accessed
April 2014]

Marketsandmarkets.com. (2011, May). Biodegradable Plastics Market: by

Types (Starch, PLA, PHA, PCL and PBS) Applications, Regulations,
Prices, Trends & Forecast (2011-2016). Retrieved from Markets and
Markets:
http://www.marketsandmarkets.com/MarketReports/biodegradable-plastics-93.html

Matija, L. (2002). Energy recovery by pinch technology, 22, 477484.

Mohan, A. M. (2011, Disember 7). World demand for bioplastics to exceed 1

million tons in 2015. Retrieved from http://www.greenerpackage.com:
http://www.greenerpackage.com/bioplastics/world_demand_bioplastic
s_exceed_1_million_tons_2015

Ng KS, Ooi WY, Goh LK, Shenbagarathai R, Sudesh K. Evaluation of

jatropha oil to produce
poly(3-hydroxybutyrate) by Curiavidus
necator H16. Polymer
Degradation
&
Stability
2010;95:1365-1369

Paramjit Singh and NitikaParmar (2011). Isolation and characterization of two

novel polyhydroxybutyrate (PHB)-producing bacteria. African
Journal of Biotechnology
Vol. 10(24), pp. 4907-4919. DOI:
10.5897/AJB10.1737

Peters M.S., Timmerhaus K.D., and West R.E. (2004). Plant Design and
Economics for
Chemical Engineers. 5th Ed. McGraw-Hill
Companies, Inc. New York.

xi

P.Rama Murthy, (2005), Production And Operations Management, New Age

International (P)
Ltd, 2nd ed, pg 181-182.

Ramsay JA, Berger E, Voyer R, Chavarie C, Ramsay BA. Extraction of poly3hydroxybutyrate

using chlorinated solvents. Biotechnol.
Techn. 1994;8:589-594

Research, B. (2012, February). Global Markets and Technologies for

Bioplastics.
Retrieved
from
bcc
research:
http://www.bccresearch.com/market-research/plastics/bioplasticsmarkets-technologies-pls050b.html

Regan MJ, Creighton JL, Desvousges WH. Sites for our solid waste: a
guidebook
for
effective
public involvement. Washington,
DC: Ofce of Solid Waste, US
Environmental
Protection
Agency; 1990

Rossiter, A. P. (2010). Improve Energy Effi ciency via Heat Integration,

(December), 3342.

Schryver, P. D. (2010). Poly--hydroxybutyrate as a microbial agent in

aquaculture. Ghent, Belgium: Ghent University. Faculty of Bioscience
Engineering.

Shen, L. H. (2011, February 9). Product overview and market projection of

emerging bio-based plastics . Retrieved from European Bioplastics:
http://www.european-bioplastics.org/index.php?id=191.

Sabra W, Abou-Zeid DM. Improving feeding strategies for maximizing

polyhydroxybutyrate yield by Bacillus megaterium. Res J Microbiol
2008;3:308-18

Salimon J, Abdullah R. Physicochemical properties of Malaysia jatropha

curcas seed oil.
Sains Malays 2008;86:553-9

xii

Shen, L. H. (2011, February 9). Product overview and market projection of

emerging bio- based plastics . Retrieved from European Bioplastics:
http://www.european- bioplastics.org/index.php?id=191.

Smriti Chand, (2014), Meaning and Objectives of Plant Layout, Industrial

Management, Retrieved
on
2
May
2014
from:
http://www.yourarticlelibrary.com/industries/meaning-and- objectivesof-plant-layoutindustrial-management/26163/

Sullivan, F. &. (2008, June 18). Frost & Sullivan. Retrieved April 24, 2014,
from Frost & Sullivan Web site: http://www.frost.com/

S. Samantaray, J.K. Nayak and N. Mallick (2011). Wastewater Utilization for

Poly-bHydroxybutyrate Production by the
CyanobacteriumAulosirafertilissima in a
Recirculatory Aquaculture
System.Appl. Environ. Microbiol.2011, 77(24):8735. DOI:
10.1128/AEM.05275-11.

S. Matsumura, Y. Shimamura, K. Terayama, T. Kiyohara(1994), Effect of

molecular weight
and stereoregularity on biodegradation of
poly(vinyl alcohol) by Alcaligenes faecalis Biothech. Lett., 16 pp.
12051210

Sun, L., & Luo, X. (2011). Synthesis of multipass heat exchanger networks
based on pinch
technology. Computers & Chemical
Engineering, 35(7), 12571264.
doi:10.1016/j.compchemeng.2010.08.005

T. Sharmila, S.A. Meenakshi, K. Kandhymathy, R. Bharathidasan et al.,

(2011). Screening
and Characterisation of Polyhydroxybutyrate
Producing Bacteria from Sugar
Industry Effluents.World Journal
of Science and Technology 2011, 1(9): 22-27

Turton, R., Bailie, R. C., Whiting, W. B., Shaeiwitz, J. A., & Bhattacharyya,
D. (2001).
Analysis, Synthesis, and Design of Chemical Processes

xiii
(Fourth Edi., Vol. 40, p. 9823).
doi:10.1002/15213773(20010316)40:6<9823::AID-ANIE9823>3.3.CO;2-C

US EPA. Municipal solid waste generation, recycling, and disposal in the

United States: facts and gures for 2010. EPA-530-F-11-005.
Washington, DC: Solid

Waste and Emergency Response; 2011 (5306P), 12 pp

Yamane T, Fukunaga M, Lee YW. Increased PHB productivity by high-celldensity-fed- batch culture of Alcaligenes latus, a growth-associated
PHB producer. Biotech.
Bioengineering
1996;50:197-202

Yu-Cui Xiong, Y.-C. Y.-Y.-Q. (2010). Application of Polyhydroxyalkanoates

Nanoparticles as. Journal of Biomaterials Science 21, 127-140.

Yusoff, S. (2006). Renewable energy from palm oil-Innovation on effective

utilization of waste. Journal of Clean Product 14: 87-93

Zahid, W.M. (2007). Cost Analysis of Trickling Filtration and Activated

Sludge Plants for The Treatment of Municipal Waste. Proceedings of
the 7th Saudi Engineering Conference, College of Engineering, King
Saud University, Riyadh on 2nd to 5th December 2007.

xiv

APPENDICES