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
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
Content 01 WINTER•Background Template•Research Objective•Research Hypothesis•Experimental - Material & Method - Experimental setup•Expected Result•Research Plan
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.
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.
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.
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.
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.
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.
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.
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
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.
Background 13 WINTER-The separation methods studied for SA Templaterecovery mainly include ;-Direct crystallization,-Precipitation,-Membrane separation,-Extraction,-Chromatography,-In situ separation..
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..
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.
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.
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
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.
Research Objectives 20-To obtain a high purity of calciumsuccinate powder which facilitatessubsequent downstream processing.
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.
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.
Experimental setup 27Experimental setup for extractive fermentationof SA using external precipitation method. Calcium succinate
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 .
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.