重组假结核耶尔森氏菌L-天冬酰胺酶的产生菌构建、表达优化及纯化

Creation of a producent, optimization of expression, and purification of recombinant Yersinia pseudotuberculosis L-asparaginase.

作者信息

Sidoruk K V, Pokrovsky V S, Borisova A A, Omeljanuk N M, Aleksandrova S S, Pokrovskaya M V, Gladilina Ju A, Bogush V G, Sokolov N N

机构信息

Institute of Genetics; Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia.

出版信息

Bull Exp Biol Med. 2011 Dec;152(2):219-23. doi: 10.1007/s10517-011-1493-7.

DOI:10.1007/s10517-011-1493-7

PMID:22808465

Abstract

Recombinant E. coli strain producing Y. pseudotuberculosis Q66CJ2 (YpA) L-asparaginase II was created. Gene ansB homologue encoding Y. pseudotuberculosis IP 32953 L-asparaginase precursor was synthesized. The gene was cloned in pBad24 expression vector and expressed in E. coli BL21 (DE3) strain. Optimal conditions for the producer strain culturing were selected. An effective method for isolation and purification of the enzyme by two-staged column chromatography was developed.

摘要

构建了产生假结核耶尔森菌Q66CJ2（YpA）L-天冬酰胺酶II的重组大肠杆菌菌株。合成了编码假结核耶尔森菌IP 32953 L-天冬酰胺酶前体的ansB同源基因。该基因被克隆到pBad24表达载体中，并在大肠杆菌BL21（DE3）菌株中表达。选择了生产菌株培养的最佳条件。开发了一种通过两步柱色谱法分离和纯化该酶的有效方法。