Method for reducing NAD analogues by utilizing methanol

Abstract

The invention discloses a method for reducing NAD analogues by utilizing methanol and applications. In the method, a reducing agent is the methanol, a catalyst is the methanol dehydrogenase capable ofutilizing the methanol, and NAD analogues can be converted into the reduced states of the NAD analogues while the methanol dehydrogenase oxidizes the methanol; the method can be used for producing the reduced state NAD analogues or the reduced state analogues of deuteration and can provide reduced state coenzymes for the enzymatic reaction consuming the reduced state NAD analogues; the reduced state NAD analogues can be used as coenzymes to be applied to the reduction reaction catalyzed by enzymes such as malic enzyme ME-L310R/Q401C, D-lactic dehydrogenase DLDH-V152R and saccharomyces cerevisiae alcohol dehydrogenase, so that the wide application of the NAD analogues can be realized; the reduction method of the NAD analogues can be performed under a mild condition; and the regenerated reduced state NAD analogues can be used for preparing malic acid or lactic acid and can be used as redox power to control and regulate the metabolic intensity of the malic acid or lactic acid in microorganisms.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an enzyme-catalyzed reduction method and application of coenzyme nicotinamide adenine dinucleotide (NAD) analogues. Specifically, methanol is used as a reducing agent to decompose NAD analogues under enzyme catalysis. Converted to its reduced state, and can be used by other enzymes as a coenzyme for reduction reactions. Background technique [0002] Nicotinamide adenine dinucleotide (NAD) and its reduced state NADH are important coenzymes in the life process, participating in redox metabolism and a series of other important biochemical processes in living organisms. These coenzymes can be used in the production of chiral chemicals and in the preparation of isotopic labels. Since many oxidoreductases use NADH or NADPH as a coenzyme, any operation that changes the concentration of NAD and its redox state will have a global impact on the cell, and it is difficult to control specific oxid...

AI Technical Summary

Problems solved by technology

[0006] Although the regeneration cycle of NAD analogs is of great significance to the fields of biocatalysis and synthetic biology, there are few literatures on the efficient reduction of NAD analogs by modifying the structure of the enzyme, and there is no literature on how to modify methanol dehydrogenase to efficiently reduce NAD analogs

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