title: Molecular cloning, co-expression, and characterization of glycerol dehydratase and 1,3-propanediol dehydrogenase from Citrobacter freundii.
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.
authors: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, China. qxh@ujs.edu.cn
authors: Xianghui
authors: X
authors: Qi
authors: Wenying
authors: W
authors: Deng
authors: Fei
authors: F
authors: Wang
authors: Qi
authors: Q
authors: Guo
authors: Huayou
authors: H
authors: Chen
authors: Liang
authors: L
authors: Wang
authors: Xiang
authors: X
authors: He
authors: Ribo
authors: R
authors: Huang
mesh_terms: D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism; D016971:Citrobacter freundii / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006836:Hydro-Lyases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D006836:Hydro-Lyases; C020671:glycerol dehydratase