ISSN 2736-1756
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (1), pp. 001-006, January, 2018. © International Scholars Journals
Full Length Research Paper
Production, purification and characterisation of extracellular L-asparaginase from a soil isolate of Bacillus sp.
Vidhya Moorthy1, Aishwarya Ramalingam1, Alagarsamy Sumantha1* and Rajesh Tippapur Shankaranaya2
1Department of Biotechnology, DKM College for Women (Autonomous), Sainathapuram, Vellore 632 001, India.
2Azyme Biosciences Private Limited, 1188/20, 3rd Floor, 26th Main, 9th Block, Jaya Nagar, Bengaluru 560 069, India.
Accepted 09 November, 2017
Abstract
L-asparaginase is an anti-neoplastic agent used in lymphoblastic leukaemia chemotherapy. Soil microbial isolates were screened for potential producers of L-asparaginase using a phenol red indicator growth medium and the microbe producing the largest hydrolysis zone was selected. The isolate was characterised by biochemical tests and was found to belong to Bacillus sp. The enzyme production was carried out by submerged fermentation. Two different carbon sources, glucose and maltose were used for the enzyme production and glucose was found to be the better carbon source. The enzyme was partially purified by ammonium sulphate precipitation. Dialysis was carried out to remove the excess salt and complete purification was achieved by ion -exchange chromatography. The characterised enzyme exhibited maximal enzyme activity at pH 7 and temperature 37°C. The enzyme was activated by MgCl2 and inhibited by EDTA. Protein profiling by SDS-PAGE revealed the molecular weight of the protein to be 45 kDa.
Key words: L-asparaginase, Bacillus, submerged fermentation, enzyme, purification.