Production and characterization of alkaline extracellular lipase from newly isolated strain Aspergil

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A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori.The growth conditions and nutritional factors for lipase production by strain HB-03 were optimized,and the maximum lipase production of (45.9±2.3) U/mL was obtained at 30 ℃ and pH 7.0 after 36 h using olive oil (1%) and sucrose (0.5%) as carbon sources and combination of peptone (2%),yeast extract (0.5%) and ammonium sulfate (0.1%) as nitrogen sources.The lipase was purified to homogeneity with 10.6-fold,18.84% yield and a specific activity of I 862.2 U/mg using ammonium sulfate precipitation followed by SephadexG-75 gel filtration chromatography.The purified lipase with molecular mass of 68 ku was estimated by SDS-PAGE.The optimum pH and temperature for the purified lipase were found to be 8.5 and 40 ℃,respectively.The lipase kept more than 80% of activity in pH 7.0-10.0 and temperatures up to 45 ℃.The metal ions of Mn2+,Ba2+ significantly enhanced the lipase activity,whereas Cu2+,Fe3+ and Mg2+ strongly reduced the lipase activity.The Km and Vmax values of the purified enzyme for p-nitrophenyl palmitate were 0.13 mmol/L and 60.6 mmol/(L·min),respectively.The results show that this novel lipase has potential industrial applications.
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