Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes

Kinetics of a lipase isolated from Bacillus sp. was studied. The enzyme showed maximum activity at pH 9 and temperature 60°C. The Michaelis constant (K(M) 0.31 mM) obtained from three different plots i.e., Lineweaver-Burk, Hanes-Wolf and Hofstee, was found to be lower than already reported lipases that confirmed higher affinity of the enzyme for its substrate p-NPL (p-nitrophenyl laurate). V(max) of the enzyme was found to be 7.6 µM/mL/min. Energy of activation calculated from Arrhenius plot was found to be 20.607 kJmol(−1). Activation enthalpy (ΔH*) had negative trend and the value for the hydrolysis of p-NPL by the enzyme at optimum temperature was −2.748 kJmol(−1). Activation entropy (ΔS*) and free energy of activation (ΔG*) of the enzyme were found to be 1.468 Jmol(−1)K(−1) and −3.237 kJmol(−1), respectively at optimum temperature. Low value of Q(10) (0.04788) shows high catalytic activity of the enzyme. Mn(2+), Fe(2+) and Mg(2+) enhanced the lipase activity whereas Cu(2+), Na(+) and Co(2+) inhibited the enzyme activity. However, the enzyme activity was not affected significantly by K(+) ions. EDTA and SDS also significantly inhibited the lipase activity. Activity of the enzyme was increased in n-hexane while decreased with increase in concentration of acetone, chloroform, ethanol and isopropanol.