title: Cloning, expression, and characterization of catechol 1,2-dioxygenase from a phenol-degrading Candida tropicalis JH8 strain.
abstract: The sequence cato encoding catechol 1,2-dioxygenase from Candida tropicalis JH8 was cloned, sequenced, and expressed in Escherichia coli. The sequence cato contained an ORF of 858 bp encoding a polypeptide of 285 amino acid residues. The recombinant catechol 1,2-dioxygenase exists as a homodimer structure with a subunit molecular mass of 32 KD. Recombinant catechol 1,2-dioxygenase was unstable below pH 5.0 and stable from pH 7.0 to 9.0; its optimum pH was at 7.5. The optimum temperature for the enzyme was 30°C, and it possessed a thermophilic activity within a broad temperature range. Under the optimal conditions with catechol as substrate, the Km and Vmax of recombinant catechol 1,2-dioxygenase were 9.2 µM and 0.987 µM/min, respectively. This is the first article presenting cloning and expressing in E. coli of catechol 1,2-dioxygenase from C. tropicalis and characterization of the recombinant catechol 1,2-dioxygenase.
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 , China.|b Hubei Provincial Cooperative Innovation Center of Industrial Fermentation , Wuhan University, Wuhan , China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Long
authors.affiliation: c College of Life Sciences , Hubei University , Wuhan , China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
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 , China.|b Hubei Provincial Cooperative Innovation Center of Industrial Fermentation , Wuhan University, Wuhan , China.
authors.forename: Zhixiong
authors.initials: Z
authors.lastname: Xie
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 , China.
authors.forename: Li
authors.initials: L
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D041022:Candida tropicalis / Q000201:enzymology* / Q000378:metabolism; D050548:Catechol 1,2-Dioxygenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010636:Phenols / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013696:Temperature
chemical_list: D010636:Phenols; D006899:Mixed Function Oxygenases; D050548:Catechol 1,2-Dioxygenase; C022336:phenol 2-monooxygenase
keywords: Candida tropicalis JH8; catechol 1,2-dioxygenase; cloning and expression; enzymatic properties