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Thesis Proposal
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Extractive fermentation of succinic acid
using an external precipitation technique
การแยกกรดซัก ซิน ิก ควบคู่ก ับ กระบวนการหมัก
ด้ว ยเทคนิค การตกตะกอนนอกถัง หมัก
Asst. Prof. Apichat Boontawan
Jiraphorn Lubsungnoen M5430116
2. Content 01
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•Background
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•Research Objective
•Research Hypothesis
•Experimental
- Material & Method
- Experimental setup
•Expected Result
•Research Plan
3. Background 02
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- Currently, the interest on development of
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biodegradable plastic since global warming
problem has come.
-Succinic acid (SA)can be used as a precursor
for synthesis biodegradable plastic
- Thailand is a large producer of raw material
to produce SA.
4. Background 03
-Commercial WINTER
scale production of SA is
petroleum-derived.Template
-However, Fermentative production of SA
from renewable resource has received
increasing interest .
- SA can be produced in microbial fermentation
by various microorganism.
5. Background 04
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- In fermentation process, cost of downstream
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processing can make a very high portion in
the total production cost, more than 50% .
- Thus ,it is necessary to reduce the production
cost of fermentation process
- In addition, product inhibition is important
factor for bioprocess development .
6. Background 05
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- In situ product removal or extractive
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fermentation can be solved the problem.
-Different technique have been introduce for
recovery technique .
-Precipitation is a very interesting technique
due to its simplicity, and no does not require
additional chemical or unit operation.
.
7. Background 06
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-This work, fermentation of SA will be used commercial
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strain of SA producing bacteria.
-CaCO3 will be used to control the pH of the broth.
- Submerge ceramic microfiltration membrane will be
employed for take out the clarified broth prior to precipitation.
-As a result, C4H4CaO4 will be precipitate out ,
the remaining will then be re-circulated back into the fermenter.
-The precipitate will be harvested and washed to produce a
high purityC4H4CaO4
8. Research Objectives 07
-To study the precipitation process of calcium
succinate by using synthetic solution.
-To investigate fermentation kinetic of
Actinobacillus succinogenes ATCC 55618
using glucose as the carbon source.
9. Research Objectives 08
-To compare SA production performance in term
of yield and volumetric productivity between this
work and conventional batch fermentation
-To obtain a high purity of calcium succinate
powder which facilitates subsequent downstream
processing.
10. Research Hypothesis 09
- SA can be produced by using Actinobacillus
succinogenes ATCC 55618.
- Precipitation of calcium succinate can be
achieved in the precipitation tank.
- Product inhibition can be minimized and SA
production is significantly increased.
14. Methods 13
Fed batch fermentation
•Fed batch fermentation will be started with 1.2 L of culture
medium and the concentrated culture medium will be
gradually added.
•pH of broth will be controlled at 6.8
•Batch fermentation will be carried out first, feeding of the
concentrated culture medium will be started when amount of
glucose is nearly depleted or lower than 20 g.L-1.
15. Experimental setup 14
Experimental setup for extractive fermentation of succinic acid
using external precipitation method.
17. Analytical procedure 16
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-Reducing sugar will be measured by Dinitrosalicylic
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(DNS) method according to Miller (1995)
- Cell concentration. Cell concentration will be
determined by using UV–VIS Spectrometer.
Cell viability will be accomplished using fluorescence
microscopy.
- Quantitative analysis of organic acids will be analyzed
by HPLC .
18. Expected results 17
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1.High yield and volumetric productivity
of succinic acid will be achieved using
the external precipitation method.
2.This simple high efficiency method
can be applied to pilot scale production
of succinic acid.
20. WINTER
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THANK YOU VERY MUCH
Jiraphorn Lubsungnoen M5430116
Editor's Notes
A. succinogenes ATTC 55618 from American Type Culture Collection (USA) Glucose as carbon source Calcium carbonate as the neutralizer Autoclave 5 L Bioreactor for fermentation High performance liquid chromatography (HPLC) Glucose analyzer
Fed batch fermentation will be started with 1.2 L of culture medium and the concentrated culture medium (7 glucose concentration) will be gradually added. The pH of broth will be controlled at 6.8 throughout the fermentation period. Batch fermentation will be carried out first, feeding of the concentrated culture medium will be started when amount of glucose is nearly depleted or lower than 20 g.L -1 .
