假单胞菌甲苯单加氧酶基因的克隆表达及其在手性亚砜合成中的应用  

Cloning and Expression of Pseudomonas Toluene Monooxygenase Genes and Its Application in Synthesis of Chiral Sulfoxide

作　　者：周阳 刘莉[1] 宋宇 吴奇梅 欧国香 彭雪娇 欧刚卫[1] 杨加伟[1] Zhou Yang;Liu Li;Song Yu;Wu Qimei;Ou Guoxiang;Peng Xuejiao;Ou Gangwei;Yang Jiawei(Department of Biochemistry, Zunyi Medical University, Zunyi, 56300)

机构地区：[1]遵义医学院生物化学教研室

出　　处：《基因组学与应用生物学》2018年第4期1465-1470,共6页Genomics and Applied Biology

基　　金：国家自然科学基金(31460230);遵义医学院大学生创新创业训练计划项目(院发[2015]5027)共同资助

摘　　要：在手性亚砜类药物合成领域,利用单加氧酶催化硫醚底物生成手性亚砜中间体是最便利和绿色的新方法。但是,现阶段具有高底物耐受、高选择性和高活性的加氧酶相当匮乏。本研究从课题组前期获得的能够高效催化硫醚生成手性亚砜的假单胞菌中克隆出编码甲苯单加氧酶的两个基因,构建共表达载体进行重组表达。最后将整细胞作为催化剂催化苯甲硫醚,初步探讨其在生物催化合成苯甲亚砜中的活性。结果表明,成功获得了这两个基因的可溶性的重组蛋白。而且,该重组蛋白具有一定的催化活性,能够有效催化少量的苯甲硫醚生成苯甲亚砜。本研究将为以后更深入的利用生物酶重组蛋白催化硫醚底物生成对应的手性亚砜打下基础。In the synthesis of c hiral sulfoxide drugs, the use of monooxygenase to catalyze sulfide to form chiral sulfoxide intermediates is the most convenient and green method. However, monooxygenases with high substrate tolerance, selectivity and activity are scarce at this stage. In this study, a toluene monooxygenase from a Pseudomonas strain isolated in our previous report were cloned to construct the expression plasmids. Then,the recombinant proteins was used as biocatalyst to catalyze thioanisole to study the activity of this enzyme in the synthesis of methyl phenyl sulfoxide. The results showed that the expression vector of this gene was successfully constructed and expressed a certain amount of soluble toluene monooxygenase recombinant proteins. The results also showed that the recombinant protein had the catalytic activity on biotransformation from methyl phenyl thioether to methyl phenyl sulfoxide. This study would contribute to use recombinant enzyme to synthesize chiral sulfoxides.

关 键 词：甲苯单加氧酶 手性亚砜 生物催化 

分 类 号：Q78[生物学—分子生物学] Q936