Abstract
1,3-Propanediol (1,3-PD), an important material for chemical industry, is biologically synthesized by glycerol dehydratase (GDHt) and 1,3-propanediol dehydrogenase (PDOR). In present study, the dhaBCE and dhaT genes encoding glycerol dehydratase and 1,3-propanediol dehydrogenase respectively were cloned from Citrobacter freundii and co-expressed in E. coli. Sequence analysis revealed that the cloned genes were 85 and 77 % identical to corresponding gene of C. freundii DSM 30040 (GenBank No. U09771), respectively. The over-expressed recombinant enzymes were purified by nickel-chelate chromatography combined with gel filtration, and recombinant GDHt and PDOR were characterized by activity assay, kinetic analysis, pH, and temperature optimization. This research may form a basis for the future work on biological synthesis of 1,3-PD.
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Acknowledgments
We thank Prof. Yong Wang of Jiangsu University for his revision to the manuscript. This study was supported by the National Natural Science Foundation of China under Grant No. 21006041, National “973” project (2011CBA00801), Scientific Research Promotion Fund for the Talents of Jiangsu University under Grant No. 08JDG009, 2010094, and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Qi, X., Deng, W., Wang, F. et al. Molecular Cloning, Co-expression, and Characterization of Glycerol Dehydratase and 1,3-Propanediol Dehydrogenase from Citrobacter freundii . Mol Biotechnol 54, 469–474 (2013). https://doi.org/10.1007/s12033-012-9585-9
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DOI: https://doi.org/10.1007/s12033-012-9585-9