Deregulation of feedback inhibition of phosphoenolpyruvate carboxylase for improved lysine production in Corynebacterium glutamicum

Applied and Environmental Microbiology
Zhen ChenAn-Ping Zeng

Abstract

Allosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition of Corynebacterium glutamicum PEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations (N917G) into the ppc gene, encoding PEPC of the lysine-producing strain C. glutamicum LC298, resulted in ∼37% improved lysine production. In vitro enzyme assays and (13)C-based metabolic flux analysis showed ca. 20 and 30% increases in the PEPC activity and corresponding flux, respectively, in the mutant strain. Higher demand for NADPH in the mutant strain increased the flux toward pentose phosphate pathway, which increased the supply of NADPH for enhanced lysine production. The present study highlights the importance of allosteric regulation on the flux control of central metabolism. The strategy described here can also be implemented to improve other oxaloacetate-derived products.

References

Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·S G GrantD Hanahan
Feb 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·Y KaiK Izui
Aug 11, 2000·Metabolic Engineering·B Christensen, J Nielsen
Dec 29, 2004·FEMS Microbiology Letters·Junko OhnishiMasato Ikeda
Aug 17, 2005·FEMS Microbiology Reviews·Uwe Sauer, Bernhard J Eikmanns
Mar 1, 2006·Journal of Industrial Microbiology & Biotechnology·Masato IkedaSatoshi Mitsuhashi
Mar 15, 2008·Microbial Cell Factories·Judith BeckerChristoph Wittmann
Mar 15, 1993·Biotechnology and Bioengineering·J J Vallino, G Stephanopoulos
Nov 6, 2008·Biotechnology and Bioengineering·Kjeld Raunkjaer Kjeldsen, Jens Nielsen
May 30, 2009·Nature Protocols·Nicola ZamboniUwe Sauer
Nov 1, 2012·Bioinformatics·Michael WeitzelWolfgang Wiechert

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Citations

Jan 13, 2016·Applied Microbiology and Biotechnology·Pengfei GuQingsheng Qi
Aug 19, 2015·Frontiers in Bioengineering and Biotechnology·Bastian Blombach, Ralf Takors
Aug 16, 2016·Current Opinion in Biotechnology·Zhen Chen, An-Ping Zeng
Feb 20, 2018·Biotechnology and Bioengineering·Kento TokuyamaHiroshi Shimizu
Mar 13, 2015·Applied Microbiology and Biotechnology·Lothar Eggeling, Michael Bott
Sep 24, 2019·Critical Reviews in Biotechnology·Fernanda Karine do Carmo FélixCarlos Ricardo Soccol
Feb 1, 2018·Proceedings of the National Academy of Sciences of the United States of America·Yu-Bin SuXuan-Xian Peng
Jun 15, 2020·Journal of Industrial Microbiology & Biotechnology·Ying-Yu WangWei-Guo Zhang
Jun 27, 2019·Applied Microbiology and Biotechnology·Maike KortmannMichael Bott
Nov 12, 2020·Metabolites·Yujue WangXiaoyang Su
Dec 22, 2020·The Plant Journal : for Cell and Molecular Biology·Robert J DiMarioAsaph B Cousins
Dec 5, 2018·Metabolic Engineering·Abigail N LeistraLydia M Contreras
Nov 8, 2017·Metabolic Engineering·Travis C KoroshBrian F Pfleger

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