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 .
SA thesis prop
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 warmingproblem has come.-Succinic acid (SA)can be used as a precursorfor synthesis biodegradable plastic- Thailand is a large producer of raw material to produce SA.
Background 03-Commercial WINTER scale production of SA ispetroleum-derived.Template-However, Fermentative production of SAfrom renewable resource has receivedincreasing interest .- SA can be produced in microbial fermentationby various microorganism.
Background 04 WINTER- In fermentation process, cost of downstream Template 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 .
Background 05 WINTER- In situ product removal or extractive Templatefermentation can be solved the problem.-Different technique have been introduce forrecovery technique .-Precipitation is a very interesting techniquedue to its simplicity, and no does not requireadditional chemical or unit operation..
Background 06 WINTER -This work, fermentation of SA will be used commercial Templatestrain of SA producing bacteria. -CaCO3 will be used to control the pH of the broth. - Submerge ceramic microfiltration membrane will beemployed 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 ahigh purityC4H4CaO4
Research Objectives 07-To study the precipitation process of calciumsuccinate by using synthetic solution.-To investigate fermentation kinetic ofActinobacillus succinogenes ATCC 55618using glucose as the carbon source.
Research Objectives 08-To compare SA production performance in termof yield and volumetric productivity between thiswork and conventional batch fermentation-To obtain a high purity of calcium succinatepowder which facilitates subsequent downstreamprocessing.
Research Hypothesis 09- SA can be produced by using Actinobacillussuccinogenes ATCC 55618.- Precipitation of calcium succinate can beachieved in the precipitation tank.- Product inhibition can be minimized and SAproduction is significantly increased.
Methods 13Fed batch fermentation •Fed batch fermentation will be started with 1.2 L of culturemedium and the concentrated culture medium will begradually added. •pH of broth will be controlled at 6.8 •Batch fermentation will be carried out first, feeding of theconcentrated culture medium will be started when amount ofglucose is nearly depleted or lower than 20 g.L-1.
Experimental setup 14Experimental setup for extractive fermentation of succinic acidusing external precipitation method.
Analytical procedure 16 WINTER-Reducing sugar will be measured by Dinitrosalicylic Template(DNS) method according to Miller (1995)- Cell concentration. Cell concentration will be determined by using UV–VIS Spectrometer. Cell viability will be accomplished using fluorescencemicroscopy.- Quantitative analysis of organic acids will be analyzed by HPLC .
Expected results 17 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.