Production, purification and characterisation of extracellular L-asparaginase from a soil isolate of Bacillus sp.


Vidhya Moorthy, Aishwarya Ramalingam, Alagarsamy Sumantha* and Rajesh Tippapur Shankaranaya

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. 

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