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Polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas stutzeri 1317 had different substrate specificities
Authors:Chen Jing-Yu  Liu Tao  Zheng Zhong  Chen Jin-Chun  Chen Guo-Qiang
Affiliation:MOE Laboratory for Protein Sciences, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, PR China.
Abstract:The whole polyhydroxyalkanoate (PHA) synthesis gene locus of Pseudomonas stutzeri strain 1317 containing PHA synthase genes phaC1Ps, phaC2Ps and PHA depolymerase gene phaZPs was cloned using a PCR cloning strategy. The sequence analysis results of the phaC1Ps, phaC2Ps and phaZPs showed high homology to the corresponding pha loci of the known Pseudomonas strains, respectively. PhaC1Ps and PhaC2Ps were functionally expressed in recombinant Escherichia coli strains and their substrate specificity was compared. The results demonstrated that PhaC1Ps and PhaC2Ps from P. stutzeri 1317 had different substrate specificities when expressed in E. coli. In details, PhaC2Ps could incorporate both short-chain-length 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates (mcl 3HA) into PHA, while PhaC1Ps only favored mcl 3HA for polymerization.
Keywords:Polyhydroxyalkanoates    PHB    PHA synthase    PCR cloning    Pseudomonas stutzeri    Escherichia coli
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