SELECTIVE SEPARATION AND PURIFICATION OF 2 LIPASES FROM CHROMOBACTERIUM-VISCOSUM USING AOT REVERSED MICELLES

Selective separation and purification of two lipases from Chromobacterium viscosum were carried out by liquid-liquid extraction using a reversed micellar system. Optimum parameters for extraction were determined using a 250 mM AOT micellar solution in isooctane. Complete separation of the two lipases was achieved at pH 6.0 with a 50 mM potassium phosphate buffer solution containing 50 mM KCl. By adding 2.5% by volume of ethanol to the lipase-loaded micellar solution, 85% of the extracted lipase could be recovered in a new aqueous phase, 50 mM K2HPO4 with 50 mM KCl, at pH 9.0. Lipase A was purified 2.6-fold with a recovery of 86%, and lipase B by 1.5-fold with a recovery of 76%.