Record Details

Cloning, expression and characterization of L-asparaginase from Pseudomonas fluorescens for large scale production in E. coli BL21.

IR@CSIR-CFTRI

View Archive Info
 
 
Field Value
 
Relation http://ir.cftri.com/12063/
http://dx.doi.org/10.1007/s13205-015-0300-y
 
Title Cloning, expression and characterization of L-asparaginase
from Pseudomonas fluorescens for large scale production
in E. coli BL21.
 
Creator Vijay, Kishore
Nishita, K. P.
Manonmani, H. K.
 
Subject 03 Enzyme Biotechnology and Engineering
 
Description L-Asparaginase (E.C. 3.5.1.1) is used as an antineoplastic
drug in the treatment of acute lymphoblastic
leukemia. L-Asparaginase from Pseudomonas fluorescens
was cloned and overexpressed in E. coli BL21. The
Enzyme was found to be a Fusion protein-asparaginase
complex which was given a lysozyme treatment and
sonication, and then was purified in a Sepharose 6B column.
The enzymatic properties of the recombinant enzyme
were studied and the kinetic parameters were determined
with kilometre of 109.99 mM and Vmax of 2.88 lM/min.
Recombinant enzyme showed pH optima at 6.3 and temperature
optima at 34 �C. Asp gene was successfully
cloned into E. coli BL21 which produced high level of
asparaginase intracellularly with 85.25 % recovery of enzyme
with a specific activity of 0.94 IU/mg protein. The
enzyme was a tetramer with molecular weight of approximately
141 kDa.
 
Date 2015
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
Rights —
 
Identifier http://ir.cftri.com/12063/1/3%20Biotech%20December%202015%2C%20Volume%205%2C%20Issue%206%2C%20pp%20975-981.pdf
Vijay, Kishore and Nishita, K. P. and Manonmani, H. K. (2015) Cloning, expression and characterization of L-asparaginase from Pseudomonas fluorescens for large scale production in E. coli BL21. 3 Biotech, 5 (6). pp. 975-981.