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

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

doi:10.1007/s10517-011-1493-7

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

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

[Article in English, Russian]

Authors

K V Sidoruk¹, V S Pokrovsky, A A Borisova, N M Omeljanuk, S S Aleksandrova, M V Pokrovskaya, Ju A Gladilina, V G Bogush, N N Sokolov

Affiliation

¹ Institute of Genetics; Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia.

PMID: 22808465
DOI: 10.1007/s10517-011-1493-7

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.

MeSH terms

Asparaginase / isolation & purification*
Asparaginase / metabolism*
Escherichia coli / enzymology
Escherichia coli / genetics
Yersinia pseudotuberculosis / enzymology*

Substances

Asparaginase