2 8 6 5 2 1 2 g
Home About us MoEF Contact us Sitemap Tamil Website  
About Envis
Whats New
Microorganisms
Research on Microbes
Database
Bibliography
Publications
Library
E-Resources
Microbiology Experts
Events
Online Submission
Access Statistics

Site Visitors

blog tracking


 
Sci Rep
Vol. 6, 2016

Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82

Noura El-Ahmady El-Naggar, Sahar F. Deraz, Hoda M. Soliman, Nehal M. El-Deeb, and Sara M. El-Ewasy

Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt.

Abstract

L-asparaginase is an important enzyme as therapeutic agents used in combination with other drugs in the treatment of acute lymphoblastic leukemia. A newly isolated actinomycetes strain, Streptomyces sp. NEAE-82, was potentially producing extracellular L-asparaginase, it was identified as Streptomyces fradiae NEAE-82, sequencing product was deposited in the GenBank database under accession number KJ467538. L-asparaginase was purified from the crude enzyme using ammonium sulfate precipitation, dialysis and ion exchange chromatography using DEAE Sepharose CL-6B. Further the kinetic studies of purified enzyme were carried out. The optimum pH, temperature and incubation time for maximum L-asparaginase activity were found to be 8.5, 40°C and 30min, respectively. The optimum substrate concentration was found to be 0.06M. The Km and Vmax of the enzyme were 0.01007M and 95.08Um-l min-l , respectively. The half-life time (T1/2) was 184.91min at 50°С, while being 179.53min at 60°С. The molecular weight of the subunits of L-asparaginase was found to be approximately 53kDa by SDS–PAGE analysis. The purified L-asparaginase showed a final specific activity of 30.636U/mg protein and was purified 3.338-fold. The present work for the first time reported more information in the production, purification and characterization of L-asparaginase produced by newly isolated actinomycetes Streptomyces fradiae NEAE-82.

 

Copyright © 2005 ENVIS Centre ! All rights reserved
This site is optimized for 1024 x 768 screen resolution