A kind of amino acid racemase and its application

An amino acid, leucine technology, applied in the direction of racemase/epimerase, enzyme, isomerase, etc., can solve problems such as narrow substrate spectrum

Active Publication Date: 2022-03-08
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the reported amino acid racemases are derived from mesophilic microorganisms with narrow substrate spectrum, such as alanine racemase, glutamate racemase, serine racemase and aspartate racemase, etc.

Method used

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  • A kind of amino acid racemase and its application
  • A kind of amino acid racemase and its application
  • A kind of amino acid racemase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Construction of recombinant expression vector

[0031] The coding gene TkAAR (nucleotide sequence shown in SEQ ID NO.2) was connected to the cloning vector pUC118, and the connection product was transformed into E. coli Escherichia coli DH5α competent cells, and spread on the LB plate culture medium containing ampicillin . After picking a single colony, insert it into LB liquid medium for culture and extract the plasmid for sequencing. The one with the correct sequencing result is the cloning vector pUC118-TK1211.

[0032] The recombinant cloning vector pUC118-TK1211 and the ESS00988 vector containing a strong promoter Pcsg were treated with NdeI and SalI double digestion, and the target gene and the vector after double digestion were ligated overnight at 16°C. The ligation product was transformed into Escherichia coli E.coli DH5α competent cells, and spread on LB plate medium containing ampicillin after recovery. Pick a single colony, insert it into LB liquid medi...

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PUM

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Abstract

The invention discloses an amino acid racemase and its application in catalyzing the racemization of amino acids, belonging to the field of biotechnology. The amino acid racemase is a polypeptide I having an amino acid sequence as shown in SEQ ID NO.1 or a polypeptide II having at least 70% identity with the amino acid sequence shown in SEQ ID NO.1 and having the function of catalyzing amino acid racemization. Amino acid racemase has racemization activity for various amino acid isomers such as Leu and Met, and is thermostable, so it can be applied to enzyme-catalyzed digestion of L-Leu / D-Leu, L-Met / D-Met Rotation is of great significance for the mutual transformation of amino acid enantiomers.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a thermophilic amino acid racemase and its application. Background technique [0002] Except for glycine, the 19 common amino acids in nature all contain α-carbon, and have two enantiomers of L-form and D-form. Although D-amino acid is not the basic structural unit of protein, many plants, animals, and microorganisms contain D-amino acid, which has important physiological functions. For example, D-amino acid is the structural unit of the peptidoglycan skeleton of bacterial cell wall, teichoic acid and poly-γ-glutamic acid; some D-amino acids can be used as signal molecules to regulate the growth and metabolism of bacteria, such as D-Leu in specific The environment can cause the disintegration of cell membranes; polypeptides containing D-amino acids such as D-aspartic acid, cysteine, and glutamic acid are used clinically due to their antibacterial activity. [0003] D-amino acid is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P41/00C12P13/06C12P13/12C12N9/90
CPCC12P41/00C12P13/06C12P13/12C12N9/90C12Y501/0101
Inventor 郑仁朝卢夏锋郑裕国
Owner ZHEJIANG UNIV OF TECH
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