手性醇脱氢酶与甲酸脱氢酶的融合蛋白体系的构建  

Construction of fusion protein systems consisting of a chiral alcohol dehydrogenase and a formate dehydrogenase

作　　者：吴希[1] 张翀[1] 邢新会[1] 

机构地区：[1]清华大学化学工程系,北京100084

出　　处：《化工学报》2009年第10期2562-2567,共6页CIESC Journal

基　　金：国家自然科学基金项目(20836004)~~

摘　　要：The bifunctional fusion protein systems consisting of Rhodococcus erythropolis chiral alcohol dehydrogenase(READH),Candida boidinii formate dehydrogenase(CbFDH) or maltose binding protein(MBP) were constructed to regenerate the cofactors for biocatalysis.READH originated from Rhodococcus erythropolis is an(S)-specific nicotinamide adenine dinucleotide(NADH)-dependent alcohol dehydrogenase,meanwhile,CbFDH originated from Candida boidinii is an NADH-dependent formate dehydrogenase.The strategies of the different fusion protein systems included:(1) fusion of the N terminus of READH to the C terminus of MBP,(2) fusion of the N terminus of CbFDH to the C terminus of MBP,(3) fusion of the N terminus of READH to the C terminus of CbFDH,(4) fusion of the C terminus of READH to the N terminus of CbFDH.The activities of READHs were depressed in all fusion strategies.When the N terminus of READH was fused to the C terminus of CbFDH,READH reached the highest activity,but CbFDH had no activity.In contrast,when the C terminus of READH was fused to the N terminus of CbFDH,CbFDH showed the highest activity,and both moieties displayed activities.From this study,the authors suggest that the rational design of the bifunctional fusion protein system may improve the biocatalysis efficiency by the simultaneous cofactor regeneration.The bifunctional fusion protein systems consisting of Rhodococcus erythropolis chiral alcohol dehydrogenase （READH）, Candida boidinii formate dehydrogenase （CbFDH） or maltose binding protein （MBP） were constructed to regenerate the cofactors for biocatalysis. READH originated from Rhodococcus erythropolis is an （S）-specific nicotinamide adenine dinucleotide （NADH）-dependent alcohol dehydrogenase, meanwhile, CbFDH originated from Candida boidinii is an NADH-dependent formate dehydrogenase. The strategies of the different fusion protein systems included： （1） fusion of the N terminus of READH to the C terminus of MBP, （2） fusion of the N terminus of CbFDH to the C terminus of MBP, （3） fusion of the N terminus of READH to the C terminus of CbFDH, （4） fusion of the C terminus of READH to the N terminus of CbFDH. The activities of READHs were depressed in all fusion strategies. When the N terminus of READH was fused to the C terminus of CbFDH, READH reached the highest activity, but CbFDH had no activity. In contrast, when the C terminus of READH was fused to the N terminus of CbFDH, CbFDH showed the highest activity, and both moieties displayed activities. From this study, the authors suggest that the rational design of the bifunctional fusion protein system may improve the biocatalysis efficiency by the simultaneous cofactor regeneration.

关 键 词：融合蛋白 辅酶再生 手性醇脱氢酶 甲酸脱氢酶 麦芽糖结合蛋白 

分 类 号：Q81[生物学—生物工程]