PARTIAL PURIFICATION AND CHARACTERISATION OF L-ASPARAGINASE FROM GORDONIA SP KM067296 ISOLATED FROM LONAR LAKE (MAHARASHTRA.INDIA).

A unique extracellular L-Asparaginase or L-asparagine amino hydrolase (E.C.3.5.1.1) was studied as a potent anti-leukemic candidate. Lonar lake provides an extraordinary environment for halo-tolerant and alkali tolerant microbes. Many researches have been carried out on its microbial flora but the study on L-asparaginase producing species have not been reported very extensively and an attempt for the same is made in this study. In the present study new potential bacterial L-asparaginase producer Gordonia terrae KM067296 has been studied for enzyme production and partial purification and characterization of the enzyme was also carried out. The crude enzyme extract was partially purified using ammonium sulfate precipitation, dialysis, and gel filtration. The partially purified L-asparaginase showed specific activity of 38.61 IU/mg and was purified up to 14 fold post gel filtration and showed maximal activity against L-asparagine when incubated at pH 7.4 at 370C for 30 minutes with substrate concentration of 0.04 M. The activity of L-asparaginase was enhanced by MgCl2 and was inhibited by EDTA. The molecular weight of the enzyme was found to be approximately 12 kDa. A Line weaver Burk analysis showed a Km value of 0.061 mM and Vmax of 0.57 IU. This study revealed Gordonia terrae KM067296 as a prospective source for high yielding and highly substrate specific production of L–asparaginase.