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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%.
Esterase PE8 isolated and identified from the salt-tolerant Pelagibacterium halotolerans B2T belongs to the esterase family 6. Due to its salt tolerance, high thermostability and high chiral selectivity, PE8 is an industrially efficient pre-chiral drug The (R) -3- (4-fluorophenyl) glutaric acid monomethyl ester ((R) -3-MFG) has great potential for development, but its molecular catalytic mechanism is not clear.PE8 is expressed in E.coli PE8 protein crystals were obtained by hanging drop crystallization and the resolution of 1.66 was obtained by X-ray diffraction analysis, belonging to the space group P21 , The unit cell parameters are a = 41.772, b = 73.398, c = 66.403, β = 102.37 ° .A isomeric unit contains two protein molecules with a solvent (water) content of 35%.