ABSTRACT
Extracellular lipases of Mucor hiemalis were purified from the culture grown in shake flasks for 96 hours. The broth was saturated with ammonium sulfate to 70%, the precipitates were dissolved and the solution desalted through a Sephadex G-25 column. The crude preparation was fractionated by chromatography with DEAE-Sephadex A-50 using a linear salt gradient. The two enzyme activities as obtained by pooling active fractions were purified further by passing each of these through a Sephadex G-l50 column. Each of the enzymes, F 1 and F 2, was found to be homogeneous by SDSPAGE. Enzyme actitives were also detected on polyacramide gel rods incorporated with 0.1% olive oil as substrate. Molecular weights of lipases F 1 and F 2 as determined by gel filteration with Sephadex G-150 were found to be 45, 710 and 19, 950, while their pH optima was 8.9 and 7.6, respectively. Lipase F 1 was purified 47.85 times with specific activity of 6.7 U/mg while lipase F2 showed 42.5 times purification with specific activity of 5.95 U/mg protein