title: Genetic engineering of Pseudomonas putida KT2442 for biotransformation of aromatic compounds to chiral cis-diols.
abstract: Toluene dioxygenase (TDO) catalyzes asymmetric cis-dihydroxylations of aromatic compounds. Pseudomonas putida KT2442 (pSPM01) harboring TDO genes could effectively biotransform a wide-range of aromatic substrates into their cis-diols products. In shake-flask culture, approximately 2.7gl(-1) benzene cis-diols, 8.8gl(-1) toluene cis-diols and 6.0gl(-1) chlorobenzene cis-diols were obtained from the biotransformation process. Furthermore, vgb gene encoding Vitreoscilla hemoglobin protein (VHb) which enhances oxygen microbial utilization rate under low dissolved oxygen concentration was integrated into P. putida KT2442 genome. The oxidation ability of the mutant strain P. putida KTOY02 (pSPM01) harboring TDO gene was increased in the presence of VHb protein. As a result, approximately 3.8, 15.1 or 6.8gl(-1) different cis-diols production was achieved in P. putida KTOY02 (pSPM01) grown in shake-flasks when benzene, toluene or chlorobenzene was used as the substrate. The above results indicate that P. putida KT2442 could be used as a cell factory to biotransform aromatic compounds.
authors.affiliation: Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
authors.forename: Shao-Ping
authors.initials: SP
authors.lastname: Ouyang
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Shang-Yu
authors.initials: SY
authors.lastname: Sun
authors.forename: Jin-Chun
authors.initials: JC
authors.lastname: Chen
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Chen
mesh_terms: D000438:Alcohols / Q000737:chemistry / Q000378:metabolism*; D001711:Biotransformation; D002876:Chromosomes, Bacterial; D005798:Genes, Bacterial; D005818:Genetic Engineering; D006841:Hydrocarbons, Aromatic / Q000737:chemistry / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D009876:Operon; D010105:Oxygenases / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*