Lipase from marine Aspergillus awamori BTMFW032: Production, partial purification and application in oil effluent treatment
Lipase from marine Aspergillus awamori BTMFW032: Production, partial purification and application in oil effluent treatment
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DOI：
10.1016/j.nbt.2011.04.007
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发表时间：
2011-10-01
影响因子：
5.4
通讯作者：
Chandrasekaran, M.
中科院分区：
文献类型：
--
作者：
Basheer, Soorej M.;Chellappan, Sreeja;Chandrasekaran, M.
从海水中分离出来的海洋真菌 BTMFW032 被鉴定为泡盛曲霉 (Aspergillus awamori)，经观察发现它能产生一种胞外脂肪酶，可减少含油污水中 92% 的脂肪和油含量。在本研究中，采用响应面法对深层发酵下脂肪酶生产的培养基进行统计优化，以实现最大酶产量。含有豆粕的培养基-0.77% (w/v)； (NH4)(2)SO4-0.1M； KH2PO4-0.05M；米糠油-2% (v/v); CaCl2-0.05 M； PEG 6000-0.05% (w/v)；氯化钠-1%（w/v）；接种量-1% (v/v); pH值3.0；孵化温度 35 摄氏度和孵化期五天被确定为最大脂肪酶产量的最佳条件。经过统计建模后，优化条件下的时程实验表明，酶的产生在孵育36小时后开始，并在96小时后达到最大值（495.0 U/ml），而酶的最大比活性在108小时记录到（1164.63 U/mg蛋白质）。优化后，脂肪酶产量总体增加了 4.6 倍。通过(NH4)(2)SO4 沉淀和离子交换色谱进行部分纯化，最终产率为33.7%。该脂肪酶的分子量为 90 kDa，在 pH 7 和 40 摄氏度时具有最佳活性。结果表明了这种海洋真菌脂肪酶在生物修复中的潜在应用范围。
Marine fungus BTMFW032, isolated from seawater and identified as Aspergillus awamori, was observed to produce an extracellular lipase, which could reduce 92% fat and oil content in the effluent laden with oil. In this study, medium for lipase production under submerged fermentation was optimized statistically employing response surface method toward maximal enzyme production. Medium with soyabean meal-0.77% (w/v); (NH4)(2)SO4-0.1 M; KH2PO4-0.05 M; rice bran oil-2% (v/v); CaCl2-0.05 M; PEG 6000-0.05% (w/v); NaCl-1% (w/v); inoculum-1% (v/v); pH 3.0; incubation temperature 35 degrees C and incubation period-five days were identified as optimal conditions for maximal lipase production. The time course experiment under optimized condition, after statistical modeling, indicated that enzyme production commenced after 36 hours of incubation and reached a maximum after 96 hours (495.0 U/ml), whereas maximal specific activity of enzyme was recorded at 108 hours (1164.63 U/mg protein). After optimization an overall 4.6-fold increase in lipase production was achieved. Partial purification by (NH4)(2)SO4 precipitation and ion exchange chromatography resulted in 33.7% final yield. The lipase was noted to have a molecular mass of 90 kDa and optimal activity at pH 7 and 40 degrees C. Results indicated the scope for potential application of this marine fungal lipase in bioremediation.