Transaminase mutant and application thereof

A technology of transaminase and mutant, which is applied to the mutant of transaminase and its application field, and can solve the problem that transaminase is difficult to catalyze synthesis of large sterically hindered chiral amine compounds and the like

Active Publication Date: 2021-06-18
ASYMCHEM LAB TIANJIN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main purpose of the present invention is to provide a transaminase mutant and its application to solve the problem that transaminase is difficult to catalyze the synthesis of large sterically hindered chiral amine compounds in the prior art

Method used

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  • Transaminase mutant and application thereof
  • Transaminase mutant and application thereof
  • Transaminase mutant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] The directional mutation was carried out on the basis of the parent parent. The specific mutation sites are shown in the following table, and the catalytic activity of the mutant was tested according to the following reaction conditions:

[0085] 1 mL system includes 1 mg substrate 1, 1 mg PLP, 1 mg isopropylamine hydrochloride, 50 mg enzyme powder, pH 8.0 100 mM phosphate buffer, 50 o C reacted for 40 h.

[0086] The test results are shown in the table below:

[0087] Table 1:

[0088]

[0089] .

[0090] Note: In the above table, 0 means the conversion rate is less than 1%, + means the conversion rate is greater than or equal to 1% and less than 5%, and ++ means the conversion rate is greater than or equal to 5% and less than or equal to 10%.

Embodiment 2

[0092] On the basis of Example 1, combined mutation was performed, and the activity of the combined mutation was screened according to the same reaction conditions as Example 1. The results are shown in the following table.

[0093] Table 2:

[0094]

[0095] .

[0096] Note: In the above table, 0 means the conversion rate is less than 1%, + means the conversion rate is greater than or equal to 1% and less than 5%, ++ means the conversion rate is greater than or equal to 5% and less than 10%, and +++ means the conversion rate is greater than or equal to 10 % and less than 15%, ++++ means the conversion rate is greater than or equal to 15% and less than 20%.

Embodiment 3

[0098] On the basis of Example 2, combined mutation was carried out, and the activity of the combined mutation was screened according to the same reaction conditions as Example 1. The results are shown in the following table.

[0099] table 3:

[0100]

[0101] .

[0102] Note: 0 means conversion rate <1%, + means conversion rate is greater than or equal to 1% and less than 5%, ++ means conversion rate is greater than or equal to 5% and less than 10%, +++ means conversion rate is greater than or equal to 10% and less than 15 %, ++++ means the conversion rate is greater than or equal to 15% and less than 20%, +++++ means the conversion rate is greater than or equal to 20% and less than or equal to 40%.

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Abstract

The invention provides a transaminase mutant and application thereof. The transaminase mutant is obtained by mutation of one or more amino acids in SEQ ID NO: 2 or is obtained by mutation of conserved amino acids in the 54th-63rd and the 84th-96th sites by taking the sequence SEQ ID NO: 1 of wild type CvTA transaminase as a reference. The catalytic activity of the mutants is improved to different extents compared with that of wild transaminase, so that when the mutants are used for synthesizing chiral amine compounds, particularly synthesizing large-steric-hindrance chiral amine, the production efficiency can be improved, and the industrial production cost can be reduced. And green chemical synthesis is carried out by adopting biological enzyme, so that industrial three wastes are reduced.

Description

technical field [0001] The present invention relates to the application field of biological enzymes, in particular, to a transaminase mutant and its application. Background technique [0002] Large sterically hindered chiral amine compounds (in this application, the group next to the potential chiral carbonyl group is larger than the methyl group) is a class of optically active substances and functional molecules, which are widely used in the synthesis of drugs and ligands. chiral intermediates. However, there are few reports on the synthesis of such chiral compounds, and satisfactory results cannot be obtained. For example, the selectivity of phenyl tert-butylamino ester with a bulky sterically hindered carbonyl side is 76% (Angew. Chem. Int. Ed. 2007, 46, 4367). At present, for the asymmetric hydrogenation reaction of small steric hindrance (in this application, the group next to the potential chiral carbonyl group is H or methyl), some metal catalysts, such as ruthenium...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12P13/00C12P17/12C12P17/14
CPCC12N9/1096C12Y206/01C12P13/001C12P17/12C12P17/14C12N15/52C12Y206/01018
Inventor 洪浩詹姆斯·盖吉肖毅张娜焦学成马玉磊牟慧艳王祖建孙凯华李响赵桐曹珊
Owner ASYMCHEM LAB TIANJIN
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