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Engineering natural and noncanonical nicotinamide cofactor-dependent enzymes: design principles and technology development.

ABSTRACT: Nicotinamide cofactors enable oxidoreductases to catalyze a myriad of important reactions in biomanufacturing. Decades of research has focused on optimizing enzymes which utilize natural nicotinamide cofactors, namely nicotinamide adenine dinucleotide (phosphate) (NAD(P)⁺). Recent findings reignite the interest in engineering enzymes to utilize noncanonical cofactors, the mimetics of NAD⁺ (mNADs), which exhibit superior industrial properties in vitro and enable specific electron delivery in vivo. We compare recent advances in engineering natural versus noncanonical cofactor-utilizing enzymes, discuss design principles discovered, and survey emerging high-throughput platforms beyond the traditional 96-well plate-based methods. Obtaining mNAD-dependent enzymes remains challenging with a limited toolkit. To this end, we highlight design principles and technologies which can potentially be translated from engineering natural to noncanonical cofactor-dependent enzymes.

SUBMITTER: King E

PROVIDER: S-EPMC7744333 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor.

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Creating enzymes and self-sufficient cells for biosynthesis of the non-natural cofactor nicotinamide cytosine dinucleotide.

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Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis.

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