ABSTRACT
Actinomycetes are most valuable microbes because they are reported to produce different kinds of novel antibiotics and bioactive compounds. L-Asparaginase enzyme is a chemotherapeutic agent used in treatment of cancer. Present study reports production and characterization of purified L-Asparaginase from an actinomycetes isolate. The isolate (A-164) was found to be a potent producer of L-Asparaginase with an enzyme activity of 26.67 IU/ml. Biochemical characterization of the isolate indicated that it belongs to genus Streptomyces. Laboratory scale enzyme production studies were undertaken using standard medium. Further, enzyme was purified by salt precipitation, anion exchange chromatography and size exclusion chromatography. The purified enzyme exhibited specific activity of 390 IU/mg with 60-fold purification and yield of 42 % with molecular weight of 34 KDa. The purified enzyme demonstrated highest activity at pH 6.0 and temperature 35oC. The enzyme retained its activity till temperature of 50oC, indicating that it is a thermostable enzyme. A study on the effect of various additives on enzyme activity was also undertaken. The Km and Vmax values for the purified L-Asparaginase were 0.065 mM and 20.80 IU/ml respectively. The enzyme can further be used for healthcare and industrial applications.
Key words: Keywords: L-Asparaginase, Streptomyces sp., Biochemical characterization, Chromatography, SDS-PAGE, Kinetic studies.
|
