Production and Partial Purification Of L-Asparaginase from Bacillus Sp.

Abstract: The present study focuses on the production & partial purification of L-asparaginase enzyme from Bacillus sp. The rhizosphere soil sample was collected and screened for L-asparaginase producing bacteria on M9 media respectively using phenol red indicator growth medium. The potential isolate was selected and tested for morphological and biochemical characteristics, which confirmed Bacillus sp. The enzyme production was carried out by submerged fermentation. The different carbon and nitrogen sources were used for the enzyme production and xylulose was found to be the better carbon source, ammonium sulphate was used as a nitrogen source. The activity of crude enzyme was checked in different physical conditions of varying pH and temperature, The characterized enzyme exhibited maximal enzyme activity at pH 7 and temperature 30°C. The crude L-asparaginase was partially purified with ammonium sulfate precipitation and dialysis. Optimization of growth parameters showed significant effect on production of enzyme L- asparaginase from 2.18 to 2.92 μmol/mL/min. Thus, use of optimized M9 broth increased enzyme activity by 33.94%.

Keywords: L-asparaginase, Bacillus sp. Partial purification, submerged fermentation.