Metabolic Flux Analysis Of E.coli Mutant For Succinic Acid Production By Fermentation Enhancing And ¹³C-labeling Techniques

Fed-batch fermentations of TUQ2 were performed using complex medium under anaerobic condition. It was found that the succinate production can reach 37.442 g/L, with the yield of 0.86mol/mol glu. While comparing the extracellular products of TUQ2 and TUQ17 cultivated under batch fermentation, it was found that although TUQ17 can effectly eliminate acetate, comparing to the succinate production reach a lower lever. It was learned that the extra carbon flux was directed to lactate.This paper adopts the ~13)C labeling technique combined with Gas Chromatography -Mass Spectrometry (GC-MS) measurement technique as quantitive analysis method to identify the central metabolic flux distributions of TUQ13 under anaerobic enviroment. This project will provide a more accurate bird's view about metabolic flux distributions under yfiD gene knockout condition and will help to locate more protential gene manipulation point for strain production improvement of succinic acid. This paper covers the entire procedures about the ~13)C labling experiment, including: chemostat experiment, GC-MS measurement and FiatFlux calculating the ultimate flux.