Construction of N-carbamoylase expression genes and engineering bacteria of N-carbamoylase expression genes

Abstract

The invention provides construction of N-carbamoylase expression genes and engineering bacteria of the N-carbamoylase expression genes. A preparation method of N-carbamoylase comprises following steps: 1, screening of target bacteria is carried out; 2, extraction of bacteria total DNA is carried out; 3, amplification of the bacteria total DNA is carried out so as to obtain N-carbamoylase genes; 4, DNA segments of the N-carbamoylase genes are connected with expression vectors so as to obtain N-carbamoylase recombinant plasmids; 5, the N-carbamoylase recombinant plasmids are transferred into expression hosts, and fermental cultivation is carried out. According to the invention, amino acid sequences coded by the N-carbamoylase genes possess relatively high enzymatic activity when N-carbamoyl-D-p-hydroxyphenylglycine is taken as a substrate.

Description

technical field

[0001] The invention relates to an N-carbamoyl hydrolase expression gene and construction of engineering bacteria thereof, in particular to an N-carbamoyl-D-p-hydroxyphenylglycine hydrolase expression gene and construction of engineering bacteria thereof. Background technique

[0002] D-amino acid and its derivatives are important raw materials in the fields of medicine and food, and are widely used in the synthesis of semi-synthetic antibiotics, polypeptide hormones, pyrethroids, insecticides and sweeteners. Among them, D-p-hydroxyphenylglycine (D-pHPG) is the main side chain in the production of semi-synthetic penicillins and semi-synthetic cephalosporins, and is widely used in the synthesis of semi-synthetic broad-spectrum antibiotics amoxicillin, cefadroxil, and cefoperazone , Cefuroxime has cefadroxol and so on.

[0003] There are two types of synthetic methods for D-p-hydroxyphenylglycine (D-p-HPG), chemical resolution and enzymatic resolution. Due to...

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