Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 109, Issue 8, Pages 2070-2081Publisher
WILEY
DOI: 10.1002/bit.24486
Keywords
citrate synthase; fluxes; metabolite pools; in vivo kinetics; metabolite production
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Funding
- [0313704]
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We here developed a series of Corynebacterium glutamicum strains with gradual decreased specific citrate synthase (CS) activity and quantified in a multifaceted approach the consequences of residual activity on the transcriptome, metabolome, and fluxome level as well as on L-lysine formation and growth. We achieved an intended gradual L-lysine yield increase and recognized and overcame further new limitations in the L-lysine biosynthesis pathway to result in a strain with the highest yield reported so far when assayed under comparable conditions. As a non-intended outcome, a detailed flux analysis revealed an almost constant flux through CS at 10% remaining CS activity, whereas the metabolome data revealed an increase in the oxaloacetate and acetyl-CoA concentrations. Hence reduced CS activity is apparently efficiently buffered by increased concentrations of CS substrates, implying a certain robustness of the central metabolism in response of the imposed gene expressions. Biotechnol. Bioeng. 2012; 109:20702081. (c) 2012 Wiley Periodicals, Inc.
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