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Omega-aminotransferase mutant gene and application thereof

A transaminase and mutant technology, applied in the field of molecular biology, can solve the problems of substrate and product inhibition, enzyme substrate specificity, enantioselectivity and/or low conversion rate, etc., to achieve the effect of improving efficiency

Inactive Publication Date: 2015-09-09
ZHEJIANG UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the substrate specificity, enantioselectivity and / or conversion of commonly used enzymes are not high enough for industrial processes
In addition, one of the most important drawbacks of using transaminases to produce optically active amines is that substrate and product inhibition often occurs

Method used

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  • Omega-aminotransferase mutant gene and application thereof
  • Omega-aminotransferase mutant gene and application thereof
  • Omega-aminotransferase mutant gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] In 2 mL of 20 mM pH 8.0 phosphate buffer, add 0.25 mg ω-TA, 5 μmol trifluoroacetone, 15 μmol L-aspartic acid and 0.1 μmol PLP, react at 25 °C for 1 h, and the conversion rate is 97.0 % of trifluoropropylamine.

Embodiment 2

[0028] In 2 mL of 20 mM pH 7.5 phosphate buffer, add 0.5 mg ω-TA, 10 μmol trifluoroacetone, 30 μmol L-alanine and 0.2 μmol PLP, react at 25 °C for 2 hours, and the conversion rate is 93.0% of trifluoropropylamine.

Embodiment 3

[0030] In 10 mL of 20 mM pH 8.0 phosphate buffer, add 2 mg ω-TA, 50 μmol trifluoroacetone, 150 μmol L-aspartic acid and 1 μmol PLP, react at 25 °C for 3 h, and the conversion rate is 92.0 % of trifluoropropylamine.

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PUM

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Abstract

The invention discloses an omega-aminotransferase mutant gene and an application thereof, and belongs to the field of the molecular biological technology. The nucleotide sequence of the omega-aminotransferase mutant gene is shown as SEQ ID NO.1. The omega-aminotransferase amino acid sequence embodied by the code of the omega-aminotransferase mutant gene is shown as SEQ ID NO.2. The omega-aminotransferase mutant gene is firstly provided, and the omega- aminotransferase corresponding to the code of the omega-aminotransferase mutant gene has a high enzyme amount, so that the biological catalysis and conversion efficiency is improved in industrial production. The invention further provides the application of the omega-aminotransferase embodied by the code of the omega-aminotransferase mutant gene in preparation of trifluoro-propylamine. The trifluoro-propylamine is added to a buffer solution to serve as the substrate for the reaction, and then products of trifluoromethyl amine compounds are collected from the reaction liquid. The method for preparing the trifluoro-propylamine has the obvious advantages that the reaction conditions are mild, no pollution is caused, and the process is simple.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to an omega-transaminase mutant gene and application thereof. Background technique [0002] Optically pure chiral amines are a class of important pharmaceutical and fine chemical intermediates, and in-depth research on the asymmetric synthesis of chiral amines has great economic benefits and application value. At present, more than 70% of drugs are chiral amines and their derivatives. For example, the synthesis of neurological drugs, cardiovascular drugs, antihypertensive drugs, anti-infective drugs and vaccines uses chiral amines as intermediates. [0003] Transaminase can not only split racemic amines through kinetics, but also generate chiral amines through asymmetric synthesis of ketones. Compared with traditional chemical catalytic methods, transaminases are more attractive and competitive, and have become industrially used to produce amino acids and chiral...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12P13/00
Inventor 黄俊蒋成君程桂林楼坚
Owner ZHEJIANG UNIVERSITY OF SCIENCE AND TECHNOLOGY
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