Abstract

ISOLATION AND SCREENING OF L-ASPARAGINASE PRODUCING BACTERIA FROM AQUATIC AND TERRESTRIAL HABITATS IN THANE DISTRICT, M.S, INDIA

Rajguru Shubhangi A.* and Deshmukh Padma V.

ABSTRACT

Microbial L-asparaginase (L-asparagine aminohydrolase, EC3.5.1.1) has attracted much attention because of its use as therapeutic agent against acute lymphoblastic leukemia and certain other kinds of Cancer in man. L-asparaginase is an extracellular enzyme produced by large number of microorganisms. Bacterial L-asparaginase from E.coli, Erwinia carotovora and Erwinia chrysanthemi are effective in treating acute lymphosarcoma and lymphoblastic leukemia. However, its prolonged administration produces the corresponding antibody in man and the antibody causes an anaphylactic shock or neutralization of the drug effect. Therefore, there is a considerable need to screen newer organisms in order to obtain strains capable of producing high yields of L-asparaginase. In the present study, an attempt has been made to screen bacterial isolates obtained from different aquatic and terrestrial habitats for the production of L-asparaginase. Three aquatic habitats and five terrestrial habitats were selected for the study. A modified M9 medium containing Phenol red & L-asparagine was used for differentiating L-asparaginase producing bacterial species from non-producers. Out of 384 isolates screened from aquatic and terrestrial habitats, 211 bacterial isolates were found to be L-asparaginase positive. These cultures were screened based on the color change of the medium from yellow to pink. Potent isolates were selected on the basis of extent of enzyme production by Rapid Plate Semi-Quantitative assay. L-asparaginase producing isolates obtained from secondary screening techniques were confirmed for both Extracellular and Intracellular enzyme production. Morphological identification & Characterization studies of selected isolates were carried out.

Keywords: L-asparaginase, acute lymphoblastic leukemia, Modified M9 medium, Identification.