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《China Biotechnology》 2009-01
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Molecular Cloning,Co-expression and Characterization of dhaF and dhaG Genes Encoding Glycerol Dehydratase Reactivating Factor of Citrobacter freundii

QI Xiang-hui1,2 LIANG Tian2 CAO Bo2 LUO Zhao-fei3 WEI Yu-tuo2YANG Deng-feng4 HUANG Ri-bo4 (1 Food and Biotechnology Engineering School,Jiangsu University,Zhenjiang 212013,China)(2 Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization,Guangxi University,Nanning 530004,China)(3 RichGene Biotech.Co.Ltd.,Shijiazhuang 050000,China 4 Guangxi Academy of Sciences,Nanning 530003,China)  
1,3-propanediol(1,3-PD) is an important material for chemical industry,therefore,there is much interest in the production of 1,3-PD.The genes dhaF and dhaG encoding glycerol dehydratase reactivase were amplified by using the genomic DNA of Citrobacter freundii.Two segments about 1.8kb and 0.4kb were obtained and inserted into pMD-18T.The similarity between the cloning genes and corresponding genes of other bacteria reach to 86%.Then,dhaF and dhaG genes were inserted into pSE380 and co-expressed in E.coli.SDS-PAGE showed: recombinant reactivase was about 30 per in the whole crude protein and was purified homogeneous,and subunits of reactivase had apparent molecular masses about 63 and 12 kDa(α and β),nondenaturing PAGE analysis the molecular mass was about 150kDa.Therefore,its subunit structure was most likely α2β2.In the presence of free adenosylcobalamin,ATP,and Mg2+,the factor reactivated glycerol dehydratase from C.freundii,which had been inactivated.The study patially elucidated the mechanism of reactivase from C.freundii and improve the manufacture of 1,3-PD by the biological pathway.
【Fund】: 国家“973”计划(2004CB719606);; 国家“863”计划(2006AA0201037)资助项目
【CateGory Index】: Q78
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1 Liying Ping,Zhiqiang liu,Yaping Xue,and Yuguo Zheng Institute of Bioengineering,Zhejiang University of Technology,Hangzhou 310014,China;Cloning and expression of Lactobacillus reuteri glycerol dehydratase gene in Escherichia coli[J];Chinese Journal of Biotechnology;2009-12
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1 QI Xiang-hui LUO Zhao-fei WEI Yu-tuo CHEN Fa-zhong WANG Shan-shan HOU Shou-hai LIAO Dong-qing HUANG Ri-bo (Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization, Guangxi University Nanning 530005, China) (Sinozyme Biotech. Co. Ltd Nanning 530005, China);Cloning, Sequence Analysis and Expression of dhaT Gene from Citrobacter freundii and Purification and Property of Corresponding Recombinant Enzyme[J];China Biotechnology;2006-07
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1 CHEN Yong-sheng△,LIU Chang-jiang,Shao Jing-wei,et al (△College of Agronomy,Inner Mongolia University for Nationalities,Tongliao 028042,China);Cloning and Expression of Glycerol Dehydratase α-Subunit Gene in E.coli and Purification of Expressed Product[J];Chinese Journal of Biologicals;2008-01
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