Tang Leli, Xia Yulin, Wu Xiuling, Chen Xiaoli, Zhang Xueling, Li He
School of Basic Courses, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
School of Basic Courses, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
Gene. 2017 Aug 20;625:64-71. doi: 10.1016/j.gene.2017.04.046. Epub 2017 Apr 27.
One clone (Lip906) exhibiting lipase activity was screened from a metagenomic library by using a medium containing tricaprylin. A novel lipase gene from the inserted fragment of Lip906 was obtained by sequencing. The phylogenetic analysis of Lip906 lipase exhibited 34% and 32% homologue to lipases from Streptomyces sp. MspMP-M5 and Rhodopirellula europaea. This gene was expressed in Escherichia coli (E. coli) BL21 (DE3), and the recombinant protein was purified and characterized. The best substrate of the recombinant Lip906 lipase was p-nitrophenyl myristate (C14). The lipase expressed maximum activity at 74°C and pH7.8, and it was found to be stable at pH values and temperatures ranging from 6.0-8.0 and 4-78°C, respectively. Furthermore, the lipase was found to be highly resistant to commercial detergent, DMSO, and EDTA, whereas its activity was stimulated in the presence of methanol and ethanol at low concentrations. The lipase showed enhanced activity in the presence of Hg, whereas the presence of the metal ions Fe, Ca, Co, and Mg inhibited the activity. These beneficial characteristics of Lip906 lipase provide some advantages for its potential application in industry.
通过使用含有三辛酸甘油酯的培养基，从宏基因组文库中筛选出一个具有脂肪酶活性的克隆（Lip906）。通过测序获得了来自Lip906插入片段的一个新的脂肪酶基因。对Lip906脂肪酶的系统发育分析表明，它与链霉菌属MspMP-M5和欧洲玫瑰单胞菌的脂肪酶具有34%和32%的同源性。该基因在大肠杆菌BL21（DE3）中表达，重组蛋白被纯化并进行了特性分析。重组Lip906脂肪酶的最佳底物是肉豆蔻酸对硝基苯酯（C14）。该脂肪酶在74°C和pH7.8时表现出最大活性，并且发现在pH值6.0 - 8.0和温度4 - 78°C范围内分别保持稳定。此外，该脂肪酶对商业洗涤剂、二甲基亚砜和乙二胺四乙酸具有高度抗性，而在低浓度甲醇和乙醇存在下其活性受到刺激。该脂肪酶在汞存在下活性增强，而金属离子铁、钙、钴和镁的存在则抑制其活性。Lip906脂肪酶的这些有益特性为其在工业中的潜在应用提供了一些优势。
