Subscribe Now Subscribe Today
Science Alert
 
Blue
   
Curve Top
Asian Journal of Biochemistry
  Year: 2008 | Volume: 3 | Issue: 6 | Page No.: 337-350
DOI: 10.3923/ajb.2008.337.350
 
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail

Cloning, Purification, Characterization and Immobilization of L-asparaginase II from E. coli W3110

Magdy M. Youssef and Mohammed A. Al-Omair

Abstract:
In the present study, we report the cloning of L-asparaginase II (asnII) gene from E. coli W3110 into pGEX-2T DNA vector. The L-asparaginase II enzyme (E.C.3.5.1.1) was overexpressed in E. coli BL21(DE3) and purified to homogeneity 238.4 fold by utilizing chromatography technique on DEAE-Sepharose fast flow, Glutathione S sepharose 4B columns and thrombin. SDS-PAGE of the purified enzyme revealed that has Mr of 40 kDa. In addition, we found that the enzyme can be efficiently immobilized in calcium alginate gelatin composites. The free enzyme has an optimum pH at 7.5 but this optimum pH is shifted to 8.5 for the immobilized enzyme. The optimum temperature, for free and immobilized enzyme were 37 and 50°C, respectively. The immobilized enzyme retained most of its activity at 60°C with high stability compared with the native enzyme when incubated at 60°C for 30 min.
PDF Fulltext XML References Citation Report Citation
 RELATED ARTICLES:
  •    Identifying the Putative Interacting Partners of the Heat Shock Protein, HtpG through Phage Display Library Screening
  •    Antimicrobial and Antioxidant Activities of Artemisia abyssinica Extracts and DNA Degradation Effects
  •    Sucrose as a Crosslinking Modifier for the Preparation of Calcium Alginate Films via External Gelation
  •    Statistical Assessment of Medium Components by Factorial Design and Surface Methodology of L-Asparaginase Production by Isolated Streptomyces radiopugnans MS1 in Submerged Fermentation using Tapioca Effluent
How to cite this article:

Magdy M. Youssef and Mohammed A. Al-Omair, 2008. Cloning, Purification, Characterization and Immobilization of L-asparaginase II from E. coli W3110. Asian Journal of Biochemistry, 3: 337-350.

DOI: 10.3923/ajb.2008.337.350

URL: https://scialert.net/abstract/?doi=ajb.2008.337.350

COMMENT ON THIS PAPER
 
 
 

 

 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 

Curve Bottom