Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Metabolic Flux Distribution in a Metabolically Engineered Escherichia coli Strain Producing Succinic Acid

  • Hong, Soon-Ho (Metabolic and Biomolecular Engineering, National Research Laboratory, and Department of Chemical Engineering and BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology) ;
  • Lee, Sang-Yup (Metabolic and Biomolecular Engineering, National Research Laboratory, and Department of Chemical Engineering and BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology)
  • Published : 2000.08.01
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Abstract

Escherichia cole NZN111, which is known as a pfl ldhA double mutant strin, was metabolically engineered to produce succinic acid by overexpressing malic enzyme into the E. coli controlled by a trc promoter. Fermentation studies were carried out in a LB medium by first growing cells aerobically to an $OD_{600}$ of 5. At this point, 0.01 mM IPTG was added to induce the overexpression of malic enzyme and the agitation speed was gradually lowered. When the culture $OD_{600}$ reached 11, a complete anaerobic condition was achieved by flushing with a $CO_3-H_2$ gas mixture. When NZN111(pTrcML) was cultured at $37^{\circ}C$, the final succinic acid concentration of 2.8 g/l could be obtained after 30 h of anaerobic cultivation. The fermentation results were analyzed by the calculation of metabolic fluxes. Metaolic flux analysis showed that about 85% of phosphoenolpyruvate (PEP) was converted to pyruvate, and further converted to malic acid by malic enzyme.

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