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-0...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 1 862.2 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 mrnol/L and 60.6 mmol/（L.min）, respectively. The results show that this novel lipase has potential industrial applications.展开更多
文摘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 1 862.2 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 mrnol/L and 60.6 mmol/（L.min）, respectively. The results show that this novel lipase has potential industrial applications.
