Activity and stability of a Rhizomucor miehei lipase in hydrophobic media

The effects of detergents and organic solvents on a commercial lipase (Lipozyme) from Rhizomucor miehei were investigated. It was shown that the detergent sodium cholate is possibly an activator of the enzyme, increasing lipase activity 2.5 times (250% of the control) when the enzyme was preincubated with 7 mM cholate. Lipozyme activity was over twice as high (230% of the control) in the presence of 80 mM Tween 80 or 90 mM Tween 20 (polyoxyethylenesorbitan monolaurate), apparently through an additional emulsifying action on the substrate. Preincubation with Tween 80 (polyoxyethylenesorbitan mono-oleate) did not affect enzyme activity. In contrast, lipase activity was completely inhibited in the presence of an 8.9 mM concentration of another non-ionic detergent, Brij 58, whereas with a 16.4 mM concentration of the cationic detergent cetyltrimethylammonium bromide (CTAB), enzyme activity was reduced by 80%. Preincubation of Lipozyme with the same concentrations of Brij 58 [poly(oxyethylene)20 cetyl ether] and CTAB promoted total inactivation of the enzyme. Organic solvents had different effects on lipase activity and stability. Of the tested solvents, hexane was least deleterious to lipase activity and did not alter enzyme stability on preincubation. These results suggest that Lipozyme can be used in esterification reactions with hexane as solvent or in hydrolysis reactions with Tween 20 or Tween 80 as emulsifying agents, after pretreatment with sodium cholate.