PARTIAL PURIFICATION AND CHARACTERIZATION OF LIPASE PRODUCED BY SACCHAROMONOSPORA AZUREA AP11/18.A. R. PADHIAR AND H.A. MODI
An extra cellular lipase produced by Saccharomonospora azurea AP11/18 was purified by ammonium sulphate precipitation, ultra filtration, dialysis followed by gel exclusion chromatography on Sephacryl S-200 using Phosphate buffer (pH 8.0). The lipase is made up of two polypeptide chain having molecular weight 50kDa and 42kDa as determined by gel filtration and by SDS- polyacrylamide gel electrophoresis. The Km and Vmax values of lipase were found to be 21.5 μMoles and 17.85 U/mg respectively. The purified lipase was active within the pH range of 6.0-13.0, with an optimum pH of 11.0, and within the temperature range of 40-80°C, with optimum temperature for the hydrolysis of pNPP at 50°C. The hydrolytic activity of the enzyme was enhanced by Mn+2 but strongly inhibited by heavy metals Hg+2 as well as EDTA. While no effect in the presence of Cu+2 and Ca+2 salts.
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