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ULTRAFILTRATION MEMBRANE:
EFFECT OF pH AND IONIC STRENGTH ON FLUX AND REJECTION
APRIL 2009
Signature
: ..................................................
Name of Candidate
Date
: 2 APRIL 2009
iii
Beloved parents;
Hj. Harun bin Jusoh & Hajjah Aminah binti Hj. Othman
iv
ACKNOWLEDGEMENT
I would to express gratitude to all who gave me the possibility to complete this
Undergraduate Research Project (PSM). I want to thank the first and foremost, my
sincere appreciation to my Undergraduate Research Project supervisor, Dr Mimi
Sakinah Binti Abdul Munaim, for guiding and encouraging me throughout this
experiment. Thanks a lot for giving me a professional training, advice and suggestion
to bring this Undergraduate Research Project to its final form. Without her support
and interest, this PSM would not have been the same as presented here.
Special appreciation for Miss Kamariah bt Mat Peah from Faculty of Civil &
Earth Resources as her interest to help for using Total Orgnanic Carbon.
And last, but not least I thank my mothers and other family members for their
continuous support while completing this PSM.
ABSTRACT
There are various methods to separate between macro molecule and non
dissolved particle in chemical process. One of the methods is separation process using
ultrafiltarion membrane. In filtration process, the macromolecules such as enzyme
will be retained on the membrane surface. This experiment is study about fouling
characteristic occur in an industry. The fouled membrane surface problem gives the
high cost operation and reduces the quality of production. Therefore, the main
objectives for this experiment are to determine the effect of pH and ionic strength on
membrane flux and rejection during fructosyltransferase (FTase) separation. The 50
kDa molecular weight cut off (MWCO) of ultrafiltration membrane was used during
this experiment. Cross flow filtration was used to run this experiment in the lab scale.
Total organic carbon (TOC) was used to analysis the concentration of sample.
Potassium dihydrogen phosphate (KH2PO4) and dipotassium hydrogen phosphate
(K2HPO4) buffer solution range pH 5 to pH 8 was applied to find the effect of pH and
various molarities of NaCl (0.5M to 2.0M) was used to find the effect of ionic
strength in ultrafiltration membrane. The experimental result shows that the optimum
pH and ionic strength was 8.0 and 0.5M, respectively, in order to separate the FTase
solution using ultrafiltration membrane.
vi
ABSTRAK
Pelbagai langkah dan teknik digunakan bagi pemisahan antara molekul macro
dan bahan tidak terlarut dalam prosess kimia. Salah satu kaedah yang digunakan ialah
proses pemisahan dengan menggunakan penapis ultra. Dalam proses pengasingan atau
penapisan ini, molekul macro seperti enzim akan tertahan di permukaan penapis.
Dalam eksperimen ini, kajian dijalankan bagi mengenal pasti ciri ciri bahan yang
menyebabkan berlaku penyumbatan penapis dalam industri kerana ia akan memberi
kesan negatif seperti peningkatan kos operasi dan mengurangkan kualiti produk.
Objektif utama kajian ini adalah bagi mengenal pasti, kesan pH dan kekuatan ionik
kepada fluks dan bahan tertahan dengan menggunakan enzim fructosyltransferase
(FTase). Saiz penapis yang digunakan dalam eksperimen ini ialah 50 kDa. Manakala
penapis aliran songsang yang digunakan adalah berskala kecil. Total organic content
(TOC) digunakan bagi menganalisa kepekatan sampel. Kalium dihodrogen Phospahte
(KH2PO4) dan diKalium hydrogen phosphate (K2HPO4) digunakan sebagai larutan
penimbal dengan skala antara pH 5 sehingga pH 8, manakala 0.5M sehingga 2.0M
Natrium Klorida (NaCl) digunakan bagi menganalisa kekuatan ionik larutan tersebut.
Keputusan daripada kajian ini menunjukan, pH 8 dan 0.5M merupakan larutan yang
optimum untuk digunakan dalam proses penapisan dengan menggunakan penapis
ultra.
TABLE OF CONTENTS
CHAPTER
TITLE
PAGE
TITLE PAGE
DECLARATION
ii
ACKNOWLEDGEMENT
iii
ABSTRACT
ABSTRAK
vi
LIST OF TABLES
xi
LIST OF FIGURES
xvi
INTRODUCTION
1.1 Background of Study
1.3 Objective
LITERATURE REVIEW
2.1 Enzyme of Fructosyltransferase
10
10
10
11
11
12
12
13
2.5 Flowsheet
14
CHAPTER
TITLE
2.6 Process Operation
15
16
16
17
18
19
19
19
20
20
21
2.8.5.1 Temperature
21
21
21
2.9 Fouling
22
22
23
23
24
PAGE
24
25
METHODOLOGY
3.1 Overall Methodology
27
27
29
30
30
31
31
CHAPTER
TITLE
PAGE
32
for Effect of pH
3.6 Flux Analysis
33
34
34
35
37
37
Membrane
4.1.2 FTase Flux at pH 6 using Ultrafiltration
38
Membrane
4.1.3 FTase Flux at pH 7 using Ultrafiltration
39
Membrane
4.1.4 FTase Flux at pH 8 using Ultrafiltration
41
Membrane
4.1.5 Overall Flux Analysis during FTase Separation
42
at Different pH Solution
4.2 Effect of pH on Membrane Rejection
4.2.1 Rejection Analysis of FTase at Different pH
45
45
Solution
4.3 Effect of Ionic Strength on Membrane Flux
4.3.1 Flux Decline during FTase Separation at 0.5 M
47
47
NaCl
4.3.2 Flux Decline during FTase Separation at 1.0 M
48
NaCl
4.3.3 Flux Decline during FTase Separation at 1.5 M
NaCl
50
CHAPTER
TITLE
4.3.4 Flux Decline during FTase Separation at 2.0 M
PAGE
51
NaCl
4.3.5 Overall Inonic Strength Analysis during FTase
53
Separation.
56
5.2 Recommendation
57
REFERENCES
58
APPENDIX A
61
APPENDIX B
74
APPENDIX C
95
xi
LIST OF TABLES
TABLE NO.
TITLE
PAGE
2.1
13
2.2
15
Characteristics
2.3
16
2.4
17
2.5
18
3.1
31
3.2
33
4.1
46
A.1
62
A.2
63
A.3
64
A.4
65
A.5
66
A.6
67
A.7
68
A.8
69
A.9
71
A.10
71
A.11
72
A.12
73
74
xvi
LIST OF FIGURES
FIGURES NO.
TITLE
PAGE
1.1
2.1
2.2
11
2.3
Non-porous membrane
12
2.4
Carriers membrane
13
2.5
Parallel Flow
14
2.6
Series Flow
14
2.7
14
2.8
16
2.9
Spiral-Wound Schematic
17
2.10
18
2.11
19
2.12
Rtotal in membrane
23
3.1
27
3.2
28
3.3
29
3.4
Polyethersulfone membrane
30
3.5
34
4.1
37
4.2
37
4.3
38
4.4
39
4.5
40
4.6
40
4.7
41
ix
FIGURES NO.
TITLE
PAGE
4.8
42
4.9
43
different pH
4.10
43
different pH
4.11
44
4.12
44
4.13
46
4.14
47
4.15
48
4.16
49
4.17
49
4.18
50
4.19
51
4.20
52
4.21
52
4.22
54
54
55
4.25
55
C.1
96
C.2
Apparatus of Experiment
96
C.3
Apparatus of Experiment
97
C.4
Sample of Experiment
97
C.5
Sample of Experiment
98
C.6
98
CHAPTER 1
INTRODUCTION
1.1
Background of Study
2
Fructosyltransferase
is
an
enzyme
transforming
sucrose
into
1.2
Problem Statement
Many bioproducts are enzyms and there is a great demand for their separation.
Conventional techniques such as precipitation, crystallization and centrifugation can
suffer from poor selectivity of separation. The high-resolution separation techniques
such as chromatography, affinity separation and electrophoresis have a very low
throughput and produce small quantities of very pure proteins; to produce larger
amounts of proteins using these methods is expensive. (Yunos and Field, 2007)
3
membrane cleaning efficiency. Such methods include intermittent back flushing, flow
pulsation and electrical field inducement. (Muthukumaran et al., 2007)
1.3
Objectives
1.4
Scope of Study
There are few purposes doing this research. The purposes are:
i.
The membrane will be used is which have 50kDA number of molecular cut
off.
ii.
iii.
KvickTM Lab Cross-Flow System Unit was used in order to separate the
solution of DI water and FTase.
iv.
v.
4
vi.
Total Organic Carbon will be used to measured the carbon in feed and
permeate
1.5
Significant of Study
By doing this research, it is hoped can add values of FTase and membrane
ultrafiltration. The main problem to solve in this experiment is to produce maximum
the production of FTase using ultrafiltartion membrane. If the common industry used
the others membrane
rane like chromatography, affinity separation and electrophoresis to
produce the FTase, this experiment hope get better result if using the ultrafiltration
membrane system.
5
Normally during the separation process between FTase and solution, FTase
fouling will occur, this because the molecular weight of FTase not suitable with the
pore size of membrane. The important thing here is use the different value of
molecular weight and pore size.
This research also suggests using the continuous system. Hence it can reduce
the cost of operation. The price which is use as a raw material to produce
fructoligoscaride (FOS) is expensive; the continuous system is preferable due to this
problem.
CHAPTER 2
LITERATURE REVIEW
In our laboratory, we have dealt with the development and optimization of the
process of cultivation of the cells of A. pullulans with the FTase activity. The
increasing interest in prebiotic compounds opens also possibilities for small-scale use
of FTase. Isolated enzyme could be a suitable form for such purposes. For that reason,
we have also recently dealt with the downstream processing of FTase from the broth
7
obtained at the cultivation of A. pullulans. The obtained data can be used for the
design of the production process of FTase and analysis of its economic efficiency.
(Vankova, Antosova, and Polakovic, 2005)
The specific cell activity with respect to dry cell mass is a crucial factor for the
control of a cultivation run if whole cells, either free or immobilized, are used as
biocatalysts. Its value reached the maximum already in the rest day at S0 = 50 g dm3
or in the second day at S0 = 200 g dm3 and 350 g dm3. The maximum value of 8860
U g1 was reached again in the cultivation with initial sucrose concentration of 350 g
dm3. As it has been mentioned above, the initial sucrose concentration influenced the
8
amount of produced FTase whereas the cell mass produced after four cultivation days
was unelected. This result suggests that the FTase production was promoted by high
sucrose concentrations. Although other authors used different activity assay
conditions and the absolute values are not fully comparable, the FTase activities of
AP CCY 27-1-1194 are of the same order of magnitude as those published for highly
active production strains, which suggests a potential of our strain for industrial
production of fructosyltransferase (Hayashi et al., 1991).
FTase of A. pullulans occurs in the periplasmic space of cells and so the part
of the enzyme is easily released to the cultivation medium. Therefore, the recovery of
the enzyme was considered from both the harvested cells and cultivation medium.
(Vankova, Antosova and Polakovic, 2005)
10
2.2 Definition of Membrane
The transmembrane pressure is the main applied driving force (Ghosh, 2003).
Due to this applied driving force, the bulk liquid medium which is the solvent is
forced through the pores. The solvent molecules carry the solute molecules towards
the membrane and in certain case through membrane. Solute molecules might be fully
11
transmitted, partially transmitted or totally retained (or rejected) by membrane
(Ghosh, 2003).
Because the membrane must allow certain constituents to pass through, they must
have a high permeability to certain types of molecules. Membrane structures consist
of the following three basic types: