Rational design of glycerol dehydratase:Swapping the genes encoding the subunitsof glycerol dehydratase to improve enzymatic properties

作     者：QI Xianghui SUN Liang LUO Zhaofei WU Jiequn MENG Xiaolei TANG Yue WEI Yutuo HUANG Ribo 

作者机构：Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization Guangxi University Nanning 530004 China Guangxi Academy of Sciences Nanning-530003 China Sinozyme Biotech. Co. Ltd. Nanning 530004 China 

出 版 物：《Chinese Science Bulletin》 (CHINESE SCIENCE BULLETIN)

年 卷 期：2006年第51卷第24期

页      面：2977-2985页

核心收录：

学科分类：0710[理学-生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 09[农学] 071007[理学-遗传学] 070303[理学-有机化学] 0901[农学-作物学] 0703[理学-化学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：National Basic Research Developmental Project, (2004CB719606) National High Technology Research and Development Program of China, (2003AA001039) 

主　　题：甘油脱水酶 基因交换技术 异源同工酶 设计 异基因组 基因克隆 

摘      要：1,3-propanediol (1,3-PD) is an important material for chemical industry,and there has been always much interest in the production of 1,3-PD using all possible routes. The genes encoding glyc-erol dehydratase (GDHt) from Citrobacter freundii,Klebsiella pneumoniae and metagenome were cloned and expressed in E. coli. All glycerol dehy-dratases but the one from metagenome could be detected to show enzyme activities. In order to im-prove the enzymatic properties of GDHts,the genes encoding α and β-γ subunits were cloned,and the enzyme characteristics were evolved by rational de-sign based on their 3D structures which were con-structed by homology modeling. Six heteroenzymes were obtained by swapping the α subunit genes of these three different-source-derived GDHts. The pH,thermal stability and Vmax of some heteroenzymes were dramatically improved by 2―5 times compared with the wild one (GDHtKP). The GDHt cloned from metagenome,originally proved to be with no enzyme activity,was converted into active enzyme by swap-ping its subunits with other different GDHts. In addi-tion,the effect of subtle 3D structural changes on the properties of the enzyme was also observed.