Diaphorase mutant with high thermal stability, diaphorase mutant gene with high thermal stability and preparation method of diaphorase mutant

A technology with high thermal stability and diaphorase, applied in the field of bioengineering, can solve the problems of high price, low output, inconvenient large-scale production, etc., achieve the effect of wide application range of pH and reduce production cost
CN111662888AActive Publication Date: 2020-09-15北京达成生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
北京达成生物科技有限公司
Publication Date
2020-09-15

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Abstract

The invention provides a diaphorase mutant with high thermal stability, a diaphorase mutant gene with high thermal stability and a preparation method of the diaphorase mutant. The diaphorase mutant isobtained through mutation of the 122nd position amino acid in the amino acid sequence of wild-type diaphorase from glycine to aspartic acid, and the amino acid sequence of the diaphorase mutant is shown in SEQ ID NO: 1. The enzymatic properties of the diaphorase mutant are the same as the enzymatic properties of the wild-type diaphorase, the thermal stability of the diaphorase mutant is significantly higher than that of the wild-type diaphorase, and the stable range of the pH value is wide; an engineering means is adopted by the preparation method of the diaphorase mutant, genes of the diaphorase mutant are cloned into an expression vector, prokaryotic expression is achieved through transformation of host cells, and a large number of highly stable diaphorase mutants can be obtained; and alot of raw materials are provided for development of clinical diagnostic reagents, and the production cost is reduced.
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Description

technical field

[0001] The invention belongs to the field of bioengineering, and specifically relates to a diaphorase mutant with high thermal stability, a gene and a preparation method thereof. Background technique

[0002] Diaphorases (DIAphorases), referred to as DIA, also known as diaphorase, lipoamide dehydrogenase. Diaphorase plays an important role in the electron transfer system in the body. It can catalyze the dehydrogenation of dihydrolipoamide or dihydrolipoic acid to become lipoamide and lipoic acid respectively. It can also catalyze the reduction of artificial electron acceptors. Such as certain dyes, ferricyanate, quinone, etc. The two hydrogens removed can reduce FAD (flavin adenine dinucleotide) to FADH 2 , and then passed to NAD+ (nicotinamide adenine dinucleotide+) to form NADH and H + . In addition, the application of diaphorase in vitro has also been extensively studied, such as in the production of useful materials, energy-related materials, environm...

Claims

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