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Title: Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis.

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

Published In Nat Chem Biol, (2020 01)

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.

PubMed ID: 31768035 Exiting the NIEHS site

MeSH Terms: Biocatalysis; Carbon/chemistry; Chromatography, Gas; Cyclohexanones/chemistry; Escherichia coli/metabolism; Kinetics; NAD/chemistry; NADP/chemistry*; Nicotinamide Mononucleotide/chemistry*; Nicotinamide Mononucleotide/genetics; Oxidation-Reduction*; Protein Conformation; Protein Engineering; Pseudomonas putida/metabolism; Ralstonia/metabolism; Software

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