title: Cloning, expression and characterization of l-asparaginase from Withania somnifera L. for large scale production.
abstract: l-Asparaginase (E.C. 3.5.1.1) is used as a therapeutic agent in the treatment of acute childhood lymphoblastic leukemia. It is found in a variety of organisms such as microbes, plants and mammals. In plants, l-asparaginase enzymes are required to catalyze the release of ammonia from asparagine, which is the main nitrogen-relocation molecule in these organisms. An Indian medicinal plant, Withania somnifera was reported as a novel source of l-asparaginase. l-Asparaginase from W. somnifera was cloned and overexpressed in E. coli. The enzymatic properties of the recombinant enzyme were investigated and the kinetic parameters (K(m), k(cat)) for a number of substrates were determined. The kinetic parameters of selected substrates were determined at various pH and the pH- and temperature-dependence profiles were analyzed. WA gene successfully cloned into E. coli BL21 (DE3) showed high asparaginase activity with a specific activity of 17.3 IU/mg protein.
authors.affiliation: Department of Plant Biotechnology, B R D School of Biosciences, Sardar Patel Maidan, Vadtal Road, Satellite Campus, Sardar Patel University, Post Box No. 39, Vallabh Vidyanagar, 388 120 Gujarat India.
authors.forename: Vishal P
authors.initials: VP
authors.lastname: Oza
authors.forename: Pritesh P
authors.initials: PP
authors.lastname: Parmar
authors.forename: Darshan H
authors.initials: DH
authors.lastname: Patel
authors.forename: R B
authors.initials: RB
authors.lastname: Subramanian