Perakine reductase mutant and application thereof

Abstract

The invention provides a Perakine reductase mutant and an application thereof. The mutant is obtained by single mutation or 126-site and 127-site combined double mutation of a wild Perakine reductaseamino acid sequence at 126th site or 127th site: H126G / R / I / C / P / K / N / W / A / Y, and R127M / P / N / S / F, wherein ' / ' represents 'or '. On the basis of a crystal three-dimensional structure and sequence alignment,a site-specific saturation mutation technology is adopted to perform molecular modification on Perakine reductase, and screening is performed to obtain a mutant with selective reduction ketene doublebonds. A method for selective reduction of ketene double bonds by using the Perikine reductase mutant as a catalyst has the advantages of simple reaction steps, mild reaction conditions, environmental friendliness, high catalytic efficiency, simple enzyme purification, good stability, strong stereoselectivity and the like, and has good application prospects in the fields of ketene double bond reduction and synthesis of related saturated ketone drugs.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a Perakine reductase mutant and its application, especially the application of the Perakine reductase mutant in selective hydrogenation of α, β-unsaturated ketone double bonds. Background technique [0002] Selective reduction of carbon-carbon double bonds of α, β-unsaturated carbonyl compounds to retain carbonyl is an important means of synthesizing saturated carbonyl compounds, and has a wide range of applications in the field of medicine and chemical industry, such as hesperidin, turmeric and dandelion flavonoids and other saturated carbonyl compounds Natural medicines are currently mainly obtained by reducing the corresponding α,β-unsaturated carbonyl compounds. However, when using chemical methods to reduce the carbon-carbon double bonds of α,β-unsaturated carbonyl compounds, it is easy to compete with carbonyl reduction, and it is challenging to ach...

AI Technical Summary

Problems solved by technology

However, when using chemical methods to reduce the carbon-carbon double bonds of α,β-unsaturated carbonyl compounds, it is easy to compete with carbonyl reduction, and it is challenging to achieve high regioselectivity. Currently, chemical methods mainly use transition metals as the The catalyst realizes the selective hydrogenation reaction of the carbon-carbon double bond of α, β-unsaturated carbonyl, but there are still unsatisfactory reaction yields and selectivities, the need to use hydrogen as a hydrogen source with hidden safety, harsh reaction conditions and environmental pollution and other issues

[0003] The biocatalytic reaction relying on double bond reductase is another important way to achieve carbon-carbon double bond reduction. Compared with chemical methods, it has the advantages of high efficiency, environmental protection, high selectivity, and mild reaction. The limitations of its substrate spectrum, enzyme yield, stability, catalytic efficiency, cofactors and other factors still cannot achieve its wide application at the industrial level. Therefore, it is urgent to develop new high-efficiency double bond reductases that can be applied to industrial applications.

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