High throughput screening method of alcohol dehydrogenase

A technology of alcohol dehydrogenase and screening method, applied in the field of enzyme screening, can solve the problems of unsuitable whole cell screening, large background interference, high coenzyme cost, etc., achieve high-throughput culture and screening, low background interference, and rapid detection accurate effect
CN106047985AInactive Publication Date: 2016-10-26JIANGNAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
JIANGNAN UNIV
Publication Date
2016-10-26
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a high throughput screening method of alcohol dehydrogenase, and provides a high throughput screening method for aliphatic series or aryl-substituted ketone alcohol dehydrogenase in 96-well plates. According to the method, aliphatic series or aryl substituted ketone is used for performing a chromogenic reaction with 2,4-dinitrophenylhydrazine under alkaline conditions. A deep-hole plate is used for performing micromation cell culture, no cell wall breaking processing needs to be performed, a traditional screening method of detecting additionally-added expensive coenzyme NAD(P)H absorbance changes is broken through, and the method has the advantages of being low in cost, easy and convenient to operate, sensitive in reaction, accurate in detection, low in equipment requirement and high in universality, and the activity and reaction conversion of alcohol dehydrogenase in a whole-cell sample can be rapidly detected.
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Description

technical field

[0001] The invention relates to an enzyme screening method, in particular to a high-throughput screening method for alcohol dehydrogenase, and belongs to the field of biotechnology. Background technique

[0002] Chiral alcohols are recognized as very important and widely used intermediates in organic synthesis. Many drugs use chiral alcohols as the main building blocks, such as the antihistamine bepotastine besilate and the lipid-lowering drug Lipitor and the antiplatelet agglutinin clopidogrel. Asymmetric reduction of prochiral carbonyl compounds to synthesize chiral alcohols is the strategy with the highest atom economy and the most potential for application and development.

[0003] Alcohol dehydrogenase (ADH) is an important oxidoreductase in organisms, which can reversibly catalyze the reduction of carbonyl groups and the oxidation of hydroxyl groups, and is widely used in the fields of biocatalysis and biomedicine. Alcohol dehydrogenase can catalyze c...

Claims

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