Amino acid racemase and application thereof

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

Active Publication Date: 2020-06-23
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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  • Amino acid racemase and application thereof
  • Amino acid racemase and application thereof
  • Amino acid racemase and application thereof

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 ligated to the cloning vector pUC118, and the ligation product was transformed into Escherichia coli DH5α competent cells, and spread on LB plate medium containing ampicillin . The single bacteria were picked and then cultured in LB liquid medium and the plasmid was extracted for sequencing. The cloned vector pUC118-TK1211 was the one with correct sequencing result.

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

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PUM

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Abstract

The invention discloses an amino acid racemase and application of the amino acid racemase in amino acid racemization catalysis, and belongs to the biotechnology field. The amino acid racemase is polypeptide I of which the amino acid sequence is disclosed by SEQ ID NO.1 or polypeptide II which has at least 70% identity with the amino acid sequence disclosed by SEQ ID NO.1 and has an amino acid racemization catalysis function. The amino acid racemase has racemization activity for various amino acid isomers, including Leu, Met and the like, and also has thermal stability, and the amino acid racemase can be applied to the racemization of enzyme catalysis L-Leu / D-Leu, and L-met / D-Met, and has an important meaning for realizing 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] In addition to glycine, the 19 common amino acids in nature all contain α-carbons, with two enantiomers of L-type and D-type. Although D-amino acids are not the basic structural units of proteins, many plants, animals, and microorganisms contain D-amino acids, which have important physiological functions. For example, D-amino acids are the structural units that make up the bacterial cell wall peptidoglycan backbone, teichoic acid and poly-γ-glutamic acid; some D-amino acids can be used as signal molecules to regulate bacterial growth and metabolism, such as D-Leu in specific The environment can cause the disintegration of cell membranes; peptides containing D-amino acids such as D-aspartic acid, cysteine, glutamic acid, etc., have antibacterial activity and are used in clinical applications. [0003] D-a...

Claims

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Application Information

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Patent Type & Authority Applications(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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