Production and characterization of alkaline extracellular lipase from newly isolated strain Aspergillus awamori HB-03 |
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Authors: | Jin-lan Xia Bin Huang Zhen-yuan Nie and Wei Wang |
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Affiliation: | (1) Department of Biotechnology, Central Leather Research Institute, 600 020 Adyar, Chennai, India; |
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Abstract: | 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 °C 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 °C, respectively. The lipase
kept more than 80% of activity in pH 7.0–10.0 and temperatures up to 45 °C. The metal ions of Mn2+, Ba2+ significantly enhanced the lipase activity, whereas Cu2+, Fe3+ and Mg2+ strongly reduced the lipase activity. The K
m and V
max 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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Keywords: | alkaline lipase Aspergillus awamori purification characterization |
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