Production, purification and characterization of lipase enzyme from Bacillus species

ABSTRACT: Extracellular lipase was isolated and purified from culture broth of bacillus species. The optimum pH and temperature for activity of lipase was found to be 8.0 and 40ºC. The lipase was found to be stable in the pH range 6.5–8.5 and temperature range 30–60ºC. The enzyme was incubated with 50% organic solvents showed a good stability and enzyme with 30% and 70% showed slight stability decreases. Fe2+, Cu2+, at 10mM enhanced hydrolytic activity of the lipase. Interestingly, K+ ions resulted in a maximal increase in lipase activity but Na+, Mg2+, and Ca2+ ions showed an antagonistic effect on this enzyme. EDTA at 10mM concentration inhibited the activity of lipase but Tween80 and Triton X-100 strongly stimulated lipase activity Sodium perchlorate and SDS (10mM) decreased 98% of original activity of lipase. The lipase enzyme activity was investigated using various concentration of p-nitrophenyl palmitate (pNPP). The Vmax and Km was calculated by various kinetic plots. Keywords: Hydrolytic activity, organic solvents, antagonistic effect, sodium perchlorate, p-nitrophenyl palmitate, kinetic plots http://www.ijbst.org/Home/papers-published/ijbst-2011-volume-4-issue-