The column was eluted with phosphate buffer (0.02M, pH 7.4). Initially, 12 fractions (fraction
volumeD3ml, each) were collected using 0.02 M phosphate buffer (pH 7.4) followed by elution with 0.02M phosphate buffer (pH 7.4) containing 1M KCl. All eluted fractions were assayed both for the lipase activity as well as their protein content [11]. The fractions showing lipase activity were pooled and lyophilized. The lyophilized lipase was reconstituted in 2ml of distilled water and extensively dialyzed against 0.05 M Tris buffer (pH 7.5). The specific activity of the purified enzyme was compared with that of the initial crude enzyme and fold purification was calculated.