Present scenario has focussed that L- asparaginase has been found to be best anti leukemic agent. It has been characterised based on the assay principle hydrolyzing L-asparagine into L-aspartic acid and ammonia. Withania somnifera (L.) Dunal (Ashwagandha); Tradtional ayurvedic medicinal plant, used in treatment of various diseases, found to have good source of enzyme L-asparaginase. In present study, different cytotypes of W. somnifera were compared for ploidy level and L-asparaginase activity. L-asparaginase was extracted from diploid and tetraploid Withania somnifera plants and tetraploid was found to have three times higher activity. Enzyme was purified using ammonium sulphate precipitation, Ion exchange and gel permeation chromatography. Purified L-asparaginase revealed molecular weight 75 kDa, Km value 1.5 mM and Vmax 526.31 μM/min. Purified enzyme has 61.22 % yield with 13.87 fold increase in specific activity.
