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%.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
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
关键词
酯酶
PE8
结晶
Esterase
PE8
Crystallization
