Creation of a producent, optimization of expression, and purification of recombinant Yersinia pseudotuberculosis L-asparaginase.
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.
Institute of Genetics; Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia.
K V
KV
Sidoruk
V S
VS
Pokrovsky
A A
AA
Borisova
N M
NM
Omeljanuk
S S
SS
Aleksandrova
M V
MV
Pokrovskaya
Ju A
JA
Gladilina
V G
VG
Bogush
N N
NN
Sokolov
D001215:Asparaginase / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015011:Yersinia pseudotuberculosis / Q000201:enzymology*
D001215:Asparaginase