Laboratory of Biotechnology, Chulabhorn Research Institute, Kamphaengphet 6 Road, Bangkok 10210, Thailand.
Mol Biotechnol. 2013 Jan;53(1):55-62. doi: 10.1007/s12033-012-9518-7.
Proteus sp. SW1 was found to produce an extracellular solvent tolerant lipase. The gene, lipA, encoding a bacterial lipase, was cloned from total Proteus sp. SW1 DNA. lipA was predicted to encode a 287 amino acid protein of 31.2 kDa belonging to the Group I proteobacterial lipases. Purified His-tagged LipA exhibited optimal activity at pH 10.0 and 55°C. It was highly stable in organic solvents retaining 112% of its activity in 100% isopropanol after 24 h, and exhibited more than 200% of its initial activity upon exposure to 60% acetone, ethanol, and hexane for 18 h. Biodiesel synthesis reactions, using a single step addition of 13% an acyl acceptor ethanol, showed that LipA was highly effective at converting palm oil into biodiesel.
发现 Proteus sp. SW1 产生一种细胞外溶剂耐受脂肪酶。从 Proteus sp. SW1 的总 DNA 中克隆了编码细菌脂肪酶的基因 lipA。lipA 预测编码一种 287 个氨基酸的 31.2 kDa 蛋白，属于第 I 组细菌脂肪酶。纯化的 His 标记的 LipA 在 pH 值为 10.0 和 55°C 时表现出最佳活性。它在有机溶剂中非常稳定，在 24 小时后在 100%异丙醇中保留了 112%的活性，在暴露于 60%丙酮、乙醇和正己烷 18 小时后，其初始活性超过 200%。使用酰基接受物乙醇的单次添加 13%的生物柴油合成反应表明，LipA 非常有效地将棕榈油转化为生物柴油。
