A citric acid cycle-deficient Escherichia coli as an efficient chassis for aerobic fermentations

Hang Zhou, Yiwen Zhang, Christopher P. Long, Xuesen Xia, Yanfen Xue, Yanhe Ma (), Maciek R. Antoniewicz (), Yong Tao () and Baixue Lin ()
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Hang Zhou: Chinese Academy of Sciences
Yiwen Zhang: Chinese Academy of Sciences
Christopher P. Long: University of Delaware
Xuesen Xia: South China University of Technology
Yanfen Xue: Chinese Academy of Sciences
Yanhe Ma: Chinese Academy of Sciences
Maciek R. Antoniewicz: University of Delaware
Yong Tao: Chinese Academy of Sciences
Baixue Lin: Chinese Academy of Sciences

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: Abstract Tricarboxylic acid cycle (TCA cycle) plays an important role for aerobic growth of heterotrophic bacteria. Theoretically, eliminating TCA cycle would decrease carbon dissipation and facilitate chemicals biosynthesis. Here, we construct an E. coli strain without a functional TCA cycle that can serve as a versatile chassis for chemicals biosynthesis. We first use adaptive laboratory evolution to recover aerobic growth in minimal medium of TCA cycle-deficient E. coli. Inactivation of succinate dehydrogenase is a key event in the evolutionary trajectory. Supply of succinyl-CoA is identified as the growth limiting factor. By replacing endogenous succinyl-CoA dependent enzymes, we obtain an optimized TCA cycle-deficient E. coli strain. As a proof of concept, the strain is engineered for high-yield production of four separate products. This work enhances our understanding of the role of the TCA cycle in E. coli metabolism and demonstrates the advantages of using TCA cycle-deficient E. coli strain for biotechnological applications.

Date: 2024
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DOI: 10.1038/s41467-024-46655-4

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