WINTER             Thesis Proposal                   Template  Extractive fermentation of succinic acid  using an external...
Content          01      WINTER•Background         Template•Research Objective•Research Hypothesis•Experimental  - Materia...
Background            02               WINTER- Currently, the interest on development of                  Templatebiodegra...
Background   03WINTERTemplate
Background          04                WINTER- SA can be used as a precursor for synthesis                  Template  biode...
Background            05                 WINTER-Thailand is a large producer of raw material                    Template t...
Background            06- Commercial scale production of SA ispetroleum-derived.-However, Fermentative production of SAfro...
Background             07-SA is a common intermediate in the metabolic                  Templatepath way of several anaero...
Background   08Template
Background            09- Many different microorganism have been                Template  screened and studied for SA such...
Background              10-Bio-based SA is potential substitute for                  Templatethe current petroleum-based p...
Background                  11                 WINTER-In addition, product inhibition is important                   Templ...
Background           12                  WINTER- The cost of downstream processing can  make a very high Template total pr...
Background        13                WINTER-The separation methods studied for SA                 Templaterecovery mainly i...
Background   14WINTERTemplate
Background               15                WINTER-Formation of byproduct is a major problem                  Templatethat ...
Background             16                WINTER- The advantage of in situ process                  Templateseparation that...
Background           17   -This work, fermentation of SA will usedcommercial strain of SA producing bacteriaand using prec...
Background             18    - Submerge ceramic microfiltration membranewill be employed for take out the clarified brothp...
Research Objectives         19-To study the precipitation process ofcalcium succinate by using syntheticsolution.-To compa...
Research Objectives    20-To obtain a high purity of calciumsuccinate powder which facilitatessubsequent downstream proces...
Research Hypothesis         21- SA can be produced by usingActinobacillus succinogenes ATCC55618.- Precipitation of calciu...
Materials            22-Actinobacillus succinogenesATCC 55618- Glucose        - Calcium carbonate- Autoclave     - 5 L Bio...
Methods                        23- Fermentation will be carried out in the absence of oxygen byflushing CO2 gas at the beg...
Methods   24
Methods/ Batch fermentation   25
Methods/ Fed batch fermentation             26 - Fed batch fermentation will be started with1.2 L of culture medium and th...
Experimental setup           27Experimental setup for extractive fermentationof SA using external precipitation method.   ...
Analytical procedure   28    WINTER      Template
Analytical procedure            29               WINTER-Reducing sugar will be measured by                  TemplateDinitr...
Expected results          30           WINTER             Template1.High yield and volumetric productivityof succinic acid...
Research plan                31   WINTER    Template
WINTER      TemplateTHANK YOU VERY MUCH      Jiraphorn Lubsungnoen   M5430116
Extractive fermentation of succinic acid using an external precipitation technique
Extractive fermentation of succinic acid using an external precipitation technique
Extractive fermentation of succinic acid using an external precipitation technique
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Extractive fermentation of succinic acid using an external precipitation technique

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  • This figure show various chemical and products that can be synthesized from SA SA can be use as a precursor of many industries such as food industry, pharmaceuticals industry ,agriculture industry, surfactant ,detergent green solvent especially biodegradable plastic.
  • This figure show Catabolic pathway leading to synthesis of SA .Bacteria mainly utilize the phosphoenolpyruvate (PEP) carboxylation reaction . 5 key enzyme are responsible for SA production including PEPCK, MDH, Menz,Fm, and Frd
  • -Different technique have been introduce for recovery technique .
  • 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 .
  • Extractive fermentation of succinic acid using an external precipitation technique

    1. 1. WINTER Thesis Proposal Template Extractive fermentation of succinic acid using an external precipitation technique การแยกกรดซัก ซิน ิก ควบคู่ก ับ กระบวนการหมักด้ว ยเทคนิค การตกตะกอนนอกถัง หมัก Asst. Prof. Apichat Boontawan Jiraphorn Lubsungnoen M5430116
    2. 2. Content 01 WINTER•Background Template•Research Objective•Research Hypothesis•Experimental - Material & Method - Experimental setup•Expected Result•Research Plan
    3. 3. Background 02 WINTER- Currently, the interest on development of Templatebiodegradable plastic since global warming problem has come.- Succinic acid (SA) is promising chemical which has a wide range of applications in many industries.
    4. 4. Background 03WINTERTemplate
    5. 5. Background 04 WINTER- SA can be used as a precursor for synthesis Template biodegradable plastic such as polybutyrate succinate (PBS) and polyamide to produce bioplastics.
    6. 6. Background 05 WINTER-Thailand is a large producer of raw material Template to produce SA such as casava and sugarcane- Therefore, the country has high potential fordevelopment of SA production in order tosupport bioplastic industry.
    7. 7. Background 06- Commercial scale production of SA ispetroleum-derived.-However, Fermentative production of SAfrom renewable resource has receivedincreasing interest .- SA can be produced by microbialfermentation using various microorganism.
    8. 8. Background 07-SA is a common intermediate in the metabolic Templatepath way of several anaerobic and facultativemicroorganism .-Those organisms suitable for the efficientproduction of SA can be categorized intobacteria.
    9. 9. Background 08Template
    10. 10. Background 09- Many different microorganism have been Template screened and studied for SA such as ; - Actinobacillus succinogenes,- Anaerobiospirillum succiniciproducens, - Mannheimia succiniciproducens and - Recombinant Escherichia coli.
    11. 11. Background 10-Bio-based SA is potential substitute for Templatethe current petroleum-based process.-However, compared with petrochemical-derived SA , biological SA production is still not economically competitive .-Because of its major draw backs:low product concentration in fermentationbroth and difficult product recovery.
    12. 12. Background 11 WINTER-In addition, product inhibition is important Templatefactor for bioprocess development .-The product inhibition effect, which can bedescribed by specific growth rate equation ai  S   P  v = v max   ∏ 1 −   S (  K + S + S2 /K i )      P  m  
    13. 13. Background 12 WINTER- The cost of downstream processing can make a very high Template total production portion in the cost, more than 50% .- It is necessary to reduce the production cost of fermentation process in order to increase its competitiveness with that of petrochemical process.
    14. 14. Background 13 WINTER-The separation methods studied for SA Templaterecovery mainly include ;-Direct crystallization,-Precipitation,-Membrane separation,-Extraction,-Chromatography,-In situ separation..
    15. 15. Background 14WINTERTemplate
    16. 16. Background 15 WINTER-Formation of byproduct is a major problem Templatethat has to be solved because it reducesSA yield and productivity, while increasesthe complexity and cost of SA recovery .-In situ product removal or extractivefermentation can be solved theaforementioned problem..
    17. 17. Background 16 WINTER- The advantage of in situ process Templateseparation that can be improved the finalSA. concentration and reduces end-productinhibition.-Precipitation process is a very interestingtechnique due to its simplicity, and no doesnot require additional chemical or unitoperation.
    18. 18. Background 17 -This work, fermentation of SA will usedcommercial strain of SA producing bacteriaand using precipitation technique couple withfermentation SA . -Calcium carbonate will be used to control thepH of the broth.
    19. 19. Background 18 - Submerge ceramic microfiltration membranewill be employed for take out the clarified brothprior to precipitation. -As a result, Calcium succinate will beprecipitate out , the remaining will then bere-circulated back into the fermenter. -The precipitate will be harvested ,washedand dried to produce a high purity calciumsuccinate
    20. 20. Research Objectives 19-To study the precipitation process ofcalcium succinate by using syntheticsolution.-To compare SA production performance interm of yield and volumetric productivitybetween this work and conventional batchfermentation.
    21. 21. Research Objectives 20-To obtain a high purity of calciumsuccinate powder which facilitatessubsequent downstream processing.
    22. 22. Research Hypothesis 21- SA can be produced by usingActinobacillus succinogenes ATCC55618.- Precipitation of calcium succinate canbe achieved in the precipitation tank.- Product inhibition can be minimized andSA production is significantly increased.
    23. 23. Materials 22-Actinobacillus succinogenesATCC 55618- Glucose - Calcium carbonate- Autoclave - 5 L Bioreactor- HPLC - Glucose analyzer
    24. 24. Methods 23- Fermentation will be carried out in the absence of oxygen byflushing CO2 gas at the beginning of fermentation
    25. 25. Methods 24
    26. 26. Methods/ Batch fermentation 25
    27. 27. Methods/ Fed batch fermentation 26 - Fed batch fermentation will be started with1.2 L of culture medium and the concentratedculture 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 bestarted when amount of glucose is nearly depletedor lower than 20 g.L-1.
    28. 28. Experimental setup 27Experimental setup for extractive fermentationof SA using external precipitation method. Calcium succinate
    29. 29. Analytical procedure 28 WINTER Template
    30. 30. Analytical procedure 29 WINTER-Reducing sugar will be measured by TemplateDinitrosalicylic (DNS) method accordingto Miller (1995)- Cell concentration will be determined byusing UV–VIS Spectrometer. Cell viability will be accomplished using fluorescencemicroscopy.- Quantitative analysis of organic acids willbe analyzed by HPLC .
    31. 31. Expected results 30 WINTER Template1.High yield and volumetric productivityof succinic acid will be achieved usingthe external precipitation method.2.This simple high efficiency methodcan be applied to pilot scale productionof succinic acid.
    32. 32. Research plan 31 WINTER Template
    33. 33. WINTER TemplateTHANK YOU VERY MUCH Jiraphorn Lubsungnoen M5430116

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