MBT Fall 2014

Microalgal Bioprocessing:
Process Technologies,
Modelling and Optimization
Hector De la Hoz Siegler. PhD.
Department of Chemical and Petroleum Engineering
University of Calgary
h.siegler@ucalgary.ca

October 28th, 2014

Hector De la Hoz Siegler

Outline of Todays Lecture

I. Introduction to microalgae
What and why
Applications:
Biofuels
Nutraceuticals

II. Culturing techniques

Medium requirements
Open ponds and photobioreactors
Phototrophic and heterotrophic

III. Optimization of heterotrophic cultures

IV. Summary

Part I

INTRODUCTION TO MICROALGAL
BIOTECHNOLOGY
3

Microalgae: what are they?

Microalgae are plant-like unicellular organisms capable of
producing several end-products that can be used as, or converted
into, fuels: hydrogen, ethanol, oil, starch, lignocellulose.
The term microalgae comprises a polyphyletic group of
photosynthetic eukaryotes. Microalgae have a great capacity for
adapting to changing environmental conditions as well as using
different substrates.

Microalgae as efficient organisms

Benefits:
Highly efficient microorganisms
Nutrient flexibility
Stress adaptability
Produce and store high amounts of oil
Other valuable byproducts

Challenges
Low culture density
Slow growth: low productivity
High production cost
5

Applications
Human and
animal food

Pharma- and
nutraceuticals

CO2 Capture

Microalgae
Biofuel
s
Waste-water
treatment /
Remediation

Fine chemicals
6

Some Commercial Applications

Species/group

Product

Application areas

Production
facilities

References

Haematococcus
pluvialis /
Chlorophyta

Carotenoids,
astaxanthin

Health food, feed additives

and pharmaceuticals

Open ponds,
PBR

Del Campo et al. (2007)

Odontella aurita
/ Bacillariophyta

Fatty acids

Pharmaceuticals,
cosmetics, baby food

Open ponds

Pulz and Gross (2004)

Isochrysis
galbana /
Chlorophyta

Fatty acids

Animal nutrition

Open ponds,
PBR

Molina Grima et al.

(1994); Pulz and Gross
(2004)

Phaedactylum
tricornutum /
Bacillariophyta

Lipids, fatty
acids

Nutrition, fuel production

Open ponds,
basins, PBR

Yongmanitchai and Ward

(1991); AcienFernandez et al. (2003)

Muriellopsis sp.
/ Chlorophyta

Carotenoids,
Lutein

Health food, food

supplement, feed

Open ponds,
PBR

Blanco et al. (2007); Del

Campo et al. (2007)

Crypthecodinium
cohnii

DHA

Food additive

Fermenters
(heterotrophic)

Carvalho et al.
(2006)

Currently, applications of microalgal biotechnology are limited to niche (small) markets. Though
high value! We expect to move into large scale markets.
7

Biofuels: a renewable energy source

Energy reserves / Energy consumption

 2nd Generation: produced from lignocellulosic

materials, i.e. ethanol from wood chips,
switch grass.
3rd Generation: fuels from microalgae
4th Generation: from crops designed for fuels
in combination with highly efficient microbes.

Land required for satisfy demand

1st Generation: derived from food-crops, i.e.

ethanol from sugar cane or corn, biodiesel
from canola or soybeans.

Time to real world application

Biofuels

10

The microalgal bio-fuels portfolio

Feedstocks

Photosynthesis

CO2

Algal Biomass:

Water
Sunlight
Trace elements

Intermediates

Biodiesel

- Oil/Lipids
- Sugars/Starch

Fuels

Hydrodeoxygenation

Hydrolysis

- Lignocellulose
Excreted products:

Pyrolysis

Green Diesel

Sugars
Bio-oil

Hydrotreating

Gasoline

SynGas

- Hydrogen

Hydrogen

- Alcohols

Alcohols

11

Biodiesel from Micro-algae

Biodiesel derived from oil crops is a potential renewable and
carbon neutral alternative to petroleum fuels.
Biodiesel from oil crops, waste cooking oil and animal fat cannot
realistically satisfy the demand for transport fuels.
Crop

Oil yield
(L/Ha)

Land area needed

(M Ha)

% of existing US
cropping area

Corn

172

1540

846

Soybean

446

594

326

Canola

1190

223

122

Oil Palm

5950

45

24

136900

1.1

Microalgae
(70% oil w/w)

Microalgae

58700

4.5

2.5

Crop land requirement

by different oil crops to
replace 50% of all
transport fuel needs of
the US. Chisti (2007).
Too optimistic to be true!

(30% oil w/w)

12

Algae as a source of oil

Species

Oil content
(% dw)

Reference

Ankistrodesmus TR-87

28 40

Ben-Amotz and Tornabene (1985)

Botryococcus braunii

25 75

Sheehan et al. (1998); Banerjee et al. (2002); Metzger and Largeau (2005)

Chlorella sp.

28 32

Sheehan et al. (1998), Chisti (2007)

Chlorella protothecoides

15 55

Xu et al. (2006)

Cyclotella DI-35
Dunaliella tertiolecta
Hantzschia DI-160

42
36 42
66

Isochrysis sp.

7 33

Nannochloris

20 - 35 (6 - 63)

Sheehan et al. (1998)

Kishimoto et al. (1994); Tsukahara and Sawayama (2005)
Sheehan et al. (1998)
Sheehan et al. (1998); Valenzuela-Espinoza et al. (2002)
Ben-Amotz and Tornabene (1985); Negoro et al. (1991); Sheehan et al. (1998)
Sheehan et al. (1998); Hu et al. (2006)

Nannochloropsis

46 (31 - 68)

Nitzschia TR-114

28 50

Kyle DJ, Gladue RM. WO 91/14427 (Patent)

Phaeodactylum tricornutum

20 31

Sheehan et al. (1998), Chisti (2007)

Scenedesmus TR-84
Stichococcus

45

Sheehan et al. (1998)

33 (9 - 59)

Sheehan et al. (1998)

Tetraselmis suecica

15 32

Thalassiosira pseudonana

(21 - 31)

Sheehan et al. (1998); Zittelli et al. (2006); Chisti (2007)

Brown et al. (1996)

13

Oil and Biodiesel

Fatty Acids

Glycerol

Triglycerides:

Biodiesel Production:

14

Polyunsaturated Fatty Acids (PUFA)

Fatty acids with multiple double bonds
EPA: C20:5

DHA: C22:6

C18:3 and longer are essential: mammals cannot synthesize

C18:3. Need to take them from their diet
Multiple biological functions as signalling molecules or building
blocks
15

Microalgae as a Source of -3 PUFA

Fish oil has been used for the commercial production of EPA
and DHA.
Factors that limit fish oil as a source of -3 fatty acids include:
taste, odour and stability problems. High purification cost.
Fish obtain -3 fatty acids from their diet.
Several species of microalgae are primary producers of long
chain PUFA.
US$ 1.5 billion/year generated from the production of DHA
(Pulz and Gross, 2004).

16

PUFA proportions in Microalgae (% TFA)

Organisms

ARA (20:4)

EPA (20:5)

DHA (22:6)

Gymnodinium splendens

30

Cricosphaera elongata

28

Isochrysis galbana

15

7.5

Monodus subterraneus

4.7

33

Nannochloropsis sp.

35

Schizochytrium sp.

1.0

2.3

40.9

Chlorella minutissima

5.7

45

Hetermastrix rotundra

28

Chromonas sp.

12.0

6.6

Cryptomonas sp.

16

10

Rhodomonas sp.

8.7

4.6

Asterionella japonica

11

20

Biddulphia sinensis

24

Crypthecodinium cohnii

30

Nitzschia laevis

6.2

19.1

Phaeodactylum Tricornutum

34.5

Skeletonema costatum

29.2

17

General Process Diagram

Culture
Extraction

Solvent
recovery
Crude
Product

Harvesting

debris

Cell disrupter

Dryer
S/L Separator

18

Part II

MICROALGAL CULTURING
TECHNIQUES
19

Nutritional requirements
Depends on application
Food or health oils: food grade chemicals
Otherwise industrial chemicals or seawater / wastewater

Carbon source: CO2, sugars, acetate, ethanol

Macronutrients: Nitrogen and phosphorus
Micronutrients: Fe, Mg, Si, S, K
Traces: Ca, Mn, Zn, Co, Se, Cu, Mo
Vitamins: B1, B12, B6, B2
Seawater: Na, K, Mg, Ca, Cl, SO4, HCO3, BO3
Br, F, IO3, Li, Rb, Sr, Ba, Mo, V, Cr, As, Se
NO3, PO4, Fe, Zn, Mn, Cu, Co, Si, Ni

20

Solar radiation in Canada

Solar radiation in Alberta

Fort McMurray:
Edmonton:
Medicine Hat:

4181 MJ/m2y
4510 MJ/m2y
5221 MJ/m2y

Munich (GE):
Naples (IT):
Kuala Lumpur:
Orlando (FL):
Acapulco (MX):
Phoenix (AZ):

4044 MJ/m2y
5293 MJ/m2y
5622 MJ/m2y
5922 MJ/m2y
7261 MJ/m2y
7621 MJ/m2y

Solar radiation data taken from: U.S. Department of Energy - EnergyPlus Weather Data.
http://apps1.eere.energy.gov/buildings/energyplus/cfm/weather_data.cfm

Culturing techniques: Open Ponds

By far, the most common
production system.
Low installation cost
Lagoons or artificial ponds
High risk of contamination
Application limited to few
species (extremophiles).
Unmixed ponds: area range from
1 - 200 Ha, depth 20-30 cm
Raceway ponds are up to 1 Ha.

23

Culturing: Close Ponds and Tanks

Simpler designs similar to open
ponds, with a cover
(greenhouses).
Aim to reduce contamination
risks.
Control CO2 looses.
Tanks are usually mixed by
aeration.
Deep tanks are inefficient. Bad
light transmission.
Easy to operate, low cost.
24

Culturing: Photobioreactors
Flat Panel photobioreactor
Arizona Center for Algal Technology and Innovation

Tubular Photobioreactor - Algae and Biofuels Facility, South

Australian Research and Development Institute

Flexible plastic film Photobioreactor - Algenol, Florida

25

Culturing: Photobioreactors
Better culture control
Higher productivity, and culture
density
Minimal contamination risk
Well mixed
Excellent temperature control
Oxygen control is an issue
High capital investment
Frequent cleaning required
Cooling required
26

Heterotrophic Production of Algae

Some algae species can grow
using an organic carbon source.
Conventional bioreactors can be
used.

27

Phototrophic vs. Heterotrophic?

Oil content
(%)

Cell conc.
(g/L)

Oil Prod.
(mg/L d)

Ettlia oleoabundans

36 42

2.9

164

Griffiths et al (2009);
Li et al. (2008)

Nannochloropsis sp.

31 68

2.1

204

Rodolfi et al. (2009)

Amphora

40 51

593

Sheehan et al. (1998)

Chlorella sp.

28 32

1.1

139

Hsieh and Wu (2009)

Chlorella vulgaris

25 42

1.7

54

Liang et al. (2009)

Chlorella zofingiensis

25.8

1.9

35

Liu et al. (2010)

Chlorella zofingiensis

51.1

9.6

354

Liu et al. (2010)

Nitzschia laevis

16.5

22.1

914

Wen and Chen (2003)

S. Limacinum (DHA)

17.3

37.9

656

Chi et al. (2009)

A. protothecoides

38.3 53.0

8.4

820

Cheng et al. (2009)

A. protothecoides

50.3 57.8

51.1

3320

Xiong et al. (2008)

Phototrophic

References

Heterotrophic

Specie

28

Part III

MODEL-BASED OPTIMIZATION OF
HETEROTROPHIC ALGAL CULTURES
29

Bioprocess Optimization

Continuous /
Real-time

Process conditions
Media formulation
Strain selection
Genetic modification
30

The Objective for Optimization

Stress

Oil storing is a metabolic response to stress,

particularly nitrogen deficiency. At nitrogen
deficient conditions, algal cells over-accumulate
lipids.

Nitrogen

As nitrogen is required for protein synthesis, its

deficiency negatively affects growth and cell
functioning. Therefore, conditions that favored oil
accumulation constraint productivity.

The challenge is to maximize biomass production

while keeping a high oil content. It is necessary to
determine the nitrogen supplementation strategy
to achieve this.

31

Understanding algal growth

Nitrogen uptake
Cellular growth

Lipid production

32

An algal growth model

Cellular growth

Nitrogen uptake

Oil production

33

Macroscopic balances

34

Optimization: Problem formulation

Subject to:

35

Simulation results
Biomass productivity in
continuous cultures

Lipid productivity in
continuous cultures
36

Experimental results

37

Biomass productivity and growth rate

38

Lipid productivity and production rate

39

Oil Productivity
(g/L h)

Bacteria

Specie
E. coli (gen. modified)

25.4

0.246

Elbahoul et al (2010)

R. opacus PD630

38.4

0.171

Kurosawa et al (2010)

M. ramanniana

67.7

0.17

Hiruta et al (1997)

C. echinulata

26.9

0.07

Kosa et al (2011)

R. toruloides

67.5

0.54

Li et al. (2007)

L. starkeyi

56.0

0.04

Kosa et al. (2011)

C. curvatus

82.7

0.47

Zhang et al. (2011)

Schizochytrium sp.

30

0.096

Ganuza et al (2007)

C. vulgaris

9.7

0.12

Doucha et al. (2011)

A. protothecoides

50.3

0.14

Xiong et al. (2008)

A. protothecoides

49.4

0.43 0.84

Microalgae

Yeasts

Lipid content
(%, w/w)

Molds

Comparative study: growth on glucose

References

De la Hoz et al (2012)

40

Optimization: closing remarks

Model-based optimization of heterotrophic microalgal cultures
allowed to reach very high densities, with biomass productivity
greater than 30 g/L d, and as high as 70 g/L d.
High oil content (4060% w/w) can be sustained with a lipid
productivity around 20 g/L d.
High quality monitoring and control is essential to achieve high
productivities.
Better control / sensors = higher productivity.
41

Summary
Algae are promising organisms: highly efficient
Good source of oil: PUFA, biodiesel precursor
Algae can growth on simple inexpensive media
Several reactor types and geometry. Application will limit
reactor choice
Several successful commercial applications currently working.
A lot of research is still needed!
42