title: Identification and characterization of phenol hydroxylase from phenol-degrading Candida tropicalis strain JH8.
abstract: The gene phhY encoding phenol hydroxylase from Candida tropicalis JH8 was cloned, sequenced, and expressed in Escherichia coli. The gene phhY contained an open reading frame of 2130 bp encoding a polypeptide of 709 amino acid residues. From its sequence analysis, it is a member of a family of flavin-containing aromatic hydroxylases and shares 41% amino acid identity with phenol hydroxylase from Trichosporon cutaneum. The recombinant phenol hydroxylase exists as a homotetramer structure with a native molecular mass of 320 kDa. Recombinant phenol hydroxylase was insensitive to pH treatment; its optimum pH was at 7.6. The optimum temperature for the enzyme was 30 °C, and its activity was rapidly lost at temperatures above 60 °C. Under the optimal conditions with phenol as substrate, the K(m) and V(max) of recombinant phenol hydroxylase were 0.21 mmol·L(-1) and 0.077 μmol·L(-1)·min(-1), respectively. This is the first paper presenting the cloning and expression in E. coli of the phenol hydroxylase gene from C. tropicalis and the characterization of the recombinant phenol hydroxylase.
authors.affiliation: a College of Life Sciences, Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), State Key Laboratory of Virology, Wuhan University, Wuhan 430072, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Long
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.forename: Zhixiong
authors.initials: Z
authors.lastname: Xie
authors.forename: Li
authors.initials: L
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D041022:Candida tropicalis / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004791:Enzyme Inhibitors / Q000737:chemistry; D004795:Enzyme Stability; D004926:Escherichia coli; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010084:Oxidation-Reduction; D019800:Phenol; D010636:Phenols / Q000737:chemistry*; D020836:Protein Structure, Quaternary; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D010636:Phenols; D019800:Phenol; D006899:Mixed Function Oxygenases; C022336:phenol 2-monooxygenase
keywords: Candida tropicalis JH8; clonage et expression; cloning and expression; enzymatic properties; gène de la phénol-hydroxalase; phenol hydroxylase gene; propriétés enzymatiques