深海耐盐杆菌来源酯酶PE8的表达纯化、结晶和晶体X-ray衍射研究（英文）  

Purification,Crystallization and X-ray Diffraction Analysis of a Halotolerant Esterase PE8

作　　者：纪锐[1,2] 匡思运 霍颖异[3] 黄静[1,2] 兰星洋 许学伟[3] 李继喜[1,2] 

机构地区：[1]复旦大学生命科学学院上海市工业菌株工程技术研究中心,上海200438 [2]复旦大学遗传工程国家重点实验室,上海200438 [3]国家海洋局第二海洋研究所海洋生态系统和生物地球化学重点实验室,浙江杭州310012

出　　处：《复旦学报（自然科学版）》2016年第4期534-537,共4页Journal of Fudan University：Natural Science

基　　金：the Program for Professor of Special Appointment(Eastern Scholar)at Shanghai Institutions of Higher Learning(No.TP2014010)to LI Jixi;China Ocean Mineral Resources R&D Association(COMRA)Special Foundation(DY125-14-E-02)to XU Xuewei

摘　　要：从深海耐盐杆菌Pelagibacterium halotolerans B2T中分离鉴定的酯酶PE8隶属于酯酶第六家族.因具有耐盐、较高的热稳定性、高手性选择性,PE8是工业上高效生产手性药物前体(R)-3-(4-氟苯基)戊二酸单甲酯((R)-3-MFG)很有开发潜力的催化剂,但其分子催化机理并不清楚.PE8在大肠杆菌中体外表达,并通过镍柱亲和层析,以及凝胶过滤层析纯化,最终通过悬滴结晶方法获得了PE8蛋白晶体.晶体经过X-ray衍射分析获得1.66的分辨率,属于空间群P21,晶胞参数为a=41.772,b=73.398,c=66.403,β=102.37°.一个异构单元包含2个蛋白分子,溶剂(水)含量为35%.The esterase PE8 was firstly identified as a new member of type Ⅵ lipolytic enzyme from the marine bacterium Pelagibacterium halotolerans B2T. Due to its balotolerance, hyperthermostability, and high enantioselectivity, PE8 is a potential candidate for the highly efficient production of chiral（R）-3-MFG in industry. Here, PE8 was purified to a high homogeneity by gel filtration chromatography, and then crystallized by hanging drop method. The crystals diffracted to 1.66 A resolution, and belong to space group P21, with unit-cell parameters a=41. 772, b=73. 398, c=66. 403 A, β= 102.37°. An asymmetric unit contains two protein molecules, with a solvent containing at 35%.

关 键 词：酯酶 PE8 结晶 

分 类 号：Q518.3[生物学—生物化学]