Expression of recombinant escherichia coli L-asparaginase II, purification and characterization
EJB-Egyptian Journal of Biochemistry and Molecular Biology [The]. 2011; 29 (2): 237-256
in English
| IMEMR
| ID: emr-117193
ABSTRACT
L-Asparaginase [ASNase] is an anti-cancer [[antineoplastic] or [cytotoxic]] chemotherapy drug that is used for the treatment of acute lymphoblastic leukemia [ALL]. An efficient and economical scheme was developed for over expression and rapid purification of the Escherichia coli enzyme. The gene encoding for the Escherichia coli L-asparaginase was PCR-amplified and cloned in pGEX-4Tl expression vector. The recombinant L-asparaginase was purified to homogeneity by affinity chromatography on glutathione Sepharose column. The recombinant enzyme had an apparent MW of 152 kDa and a K[m] value of 12.5 microM for the main physiological substrate L-asparagine. The pI value was 5.6 while the turnover number [catalytic constant] was 1 x 10[2] s[-1] and the K[cat]/K[m] value [specificity constant] was 0.8 x 10[7] M[-1]s[-1]
Search on Google
Index:
IMEMR (Eastern Mediterranean)
Main subject:
Asparaginase
/
Cloning, Organism
/
Escherichia coli
/
Antineoplastic Agents
Language:
English
Journal:
Egypt. J. Biochem. Mol. Biol.
Year:
2011
Similar
MEDLINE
...
LILACS
LIS