Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis

William B. Black, Linyue Zhang, Wai Shun Mak, Sarah Maxel, Youtian Cui, Edward King, Bonnie Fong, Alicia Sanchez Martinez, Justin B. Siegel, Han Li

Research output: Contribution to journalArticle

Abstract

Biological production of chemicals often requires the use of cellular cofactors, such as nicotinamide adenine dinucleotide phosphate (NADP+). These cofactors are expensive to use in vitro and difficult to control in vivo. We demonstrate the development of a noncanonical redox cofactor system based on nicotinamide mononucleotide (NMN+). The key enzyme in the system is a computationally designed glucose dehydrogenase with a 107-fold cofactor specificity switch toward NMN+ over NADP+ based on apparent enzymatic activity. We demonstrate that this system can be used to support diverse redox chemistries in vitro with high total turnover number (~39,000), to channel reducing power in Escherichia coli whole cells specifically from glucose to a pharmaceutical intermediate, levodione, and to sustain the high metabolic flux required for the central carbon metabolism to support growth. Overall, this work demonstrates efficient use of a noncanonical cofactor in biocatalysis and metabolic pathway design.

Original languageEnglish (US)
JournalNature Chemical Biology
DOIs
StateAccepted/In press - Jan 1 2019

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Black, W. B., Zhang, L., Mak, W. S., Maxel, S., Cui, Y., King, E., Fong, B., Sanchez Martinez, A., Siegel, J. B., & Li, H. (Accepted/In press). Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis. Nature Chemical Biology. https://doi.org/10.1038/s41589-019-0402-7