Fervidobacterium changbaicum Lip1: identification, cloning, and characterization of the thermophilic lipase as a new member of bacterial lipase family V
Fervidobacterium changbaicum Lip1: identification, cloning, and characterization of the thermophilic lipase as a new member of bacterial lipase family V
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Fervidobacter changbaicum Lip1：作为细菌脂肪酶家族 V 新成员的嗜热脂肪酶的鉴定、克隆和表征
DOI：
10.1007/s00253-010-2971-y
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发表时间：
2011-03-01
影响因子：
5
通讯作者：
Feng, Yan
中科院分区：
文献类型：
--
作者：
Cai, Jingang;Xie, Yuan;Feng, Yan
利用脂肪酶探测引物和基因组步移技术，从极端嗜热菌长白铁杆菌CBS-1中克隆到一个新的脂肪酶基因，编码315个氨基酸。序列比对和系统发育分析表明，该酶为细菌脂肪酶家族V的新成员。长白菌脂肪酶1（FCLip 1）在78°C和pH7.8时具有最大活性。它在70°C下显示出极高的热稳定性，并且在6.0至12.0的宽pH范围内也是稳定的。动力学研究表明，FCLip 1优先水解中等长度的酰基链，特别是对硝基苯基癸酸酯和三辛酸甘油酯。以癸酸对硝基苯酯为底物，该酶的Km和kcat分别为4.67 μM和22.7/s。此外，FCLip 1对各种洗涤剂和有机溶剂具有抗性。该酶是首次报道的来自嗜热菌科的嗜热脂肪酶。其对温度和pH的极端稳定性，沿着其甘油三酯水解活性，表明FCLip 1具有很高的未来应用潜力。
A novel lipase gene encoded 315 amino acid residues was obtained using lipase-prospecting primers and genome walking from hyperthermophilic bacterium Fervidobacterium changbaicum CBS-1. Sequence alignment and phylogenetic analysis revealed this novel lipase is a new member of bacterial lipase family V. The recombinant enzyme F. changbaicum lipase 1 (FCLip1) showed maximum activity at 78°C and pH 7.8. It displayed extreme thermostability at 70°C and was also stable across a wide pH range from 6.0 to 12.0. Kinetic study demonstrated FCLip1 preferentially hydrolyzed middle-length acyl chains, especially p-nitrophenyl caprate and tricaprylin. With p-nitrophenyl caprate as a substrate, the enzyme exhibited a Km and kcat of 4.67 μM and 22.7/s, respectively. In addition, FCLip1 was resistant to various detergents and organic solvents. This enzyme is the first reported thermophilic lipase from bacterial family Thermotogaceae. Its extreme stability with respect to temperature and pH, along with its triglyceride hydrolysis activity, indicate that FCLip1 has high potential for future application.