title: Identification of a metagenome-derived prephenate dehydrogenase gene from an alkaline-polluted soil microorganism.
abstract: A novel prephenate dehydrogenase gene designated pdhE-1 was cloned by sequence-based screening of a plasmid metagenomic library from uncultured alkaline-polluted microorganisms. The deduced amino acid sequence comparison and phylogenetic analysis indicated that PdhE-1 and other putative prephenate dehydrogenases were closely related. The putative prephenate dehydrogenase gene was subcloned into pETBlue-2 vector and overexpressed in Escherichia coli BL21(DE3) pLacI. The recombinant protein was purified to homogeneity. The maximum activity of the PdhE-1 protein occurred at pH 8.0 and 45 °C using prephenic acid as the substrate. The prephenate dehydrogenase had an apparent K m value of 0.87 mM, a V max value of 41.5 U/mg, a k cat value of 604.8/min and a k cat/K m value of 1.16 × 10(4)/mol/s. L-Tyrosine did not obviously inhibit the recombinant PdhE-1 protein. The identification of a metagnome-derived prephenate dehydrogenase provides novel material for studies and application of proteins involved in tyrosine biosynthesis.
authors.affiliation: The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi, 530004, China. jiangcj0520@gmail.com
authors.forename: Chengjian
authors.initials: C
authors.lastname: Jiang
authors.forename: Bangqiao
authors.initials: B
authors.lastname: Yin
authors.forename: Mingze
authors.initials: M
authors.lastname: Tang
authors.forename: Gaochao
authors.initials: G
authors.lastname: Zhao
authors.forename: Jianjun
authors.initials: J
authors.lastname: He
authors.forename: Peihong
authors.initials: P
authors.lastname: Shen
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015698:Genomic Library; D007700:Kinetics; D054892:Metagenome; D008969:Molecular Sequence Data; D010802:Phylogeny; D011303:Prephenate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D012988:Soil Microbiology; D014443:Tyrosine / Q000737:chemistry / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; D014443:Tyrosine; D011303:Prephenate Dehydrogenase