title: Purification and characterization of alkylcatechol 2,3-dioxygenase from butylphenol degradation pathway of Pseudomonas putida MT4.
abstract: Alkylcatechol 2,3-dioxygenase was purified from the cell extract of recombinant Escherichia coli JM109 harboring the alkylcatechol 2,3-dioxygenase gene (bupB) cloned from the butylphenol-degrading bacterium Pseudomonas putida MT4. The purified enzyme (BupB) showed relative meta-cleavage activities for the following catechols: catechol (100%), 4-methylcatechol (572%), 4-n-butylcatechol (185%), 4-n-hexylcatechol (53%), 4-n-heptylcatechol (45%), 4-n-nonylcatechol (10%), 4-tert-butylcatechol (0%), and 3-methylcatechol (33%). The kinetic parameters, namely, K(m) and V(max), for catechol, 4-methylcatechol, and 4-n-butylcatechol, were 23.4, 8.4, and 6.5 microM and 25.8, 76.9, and 18.0 U mg(-1), respectively. These results suggest that BupB has broad substrate specificity for 4-n-alkylcatechols.
authors.affiliation: Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo, 2167 Shosha, Himeji, Hyogo 671-2201, Japan. takeo@eng.u-hyogo.ac.jp
authors.forename: Masahiro
authors.initials: M
authors.lastname: Takeo
authors.forename: Munehiro
authors.initials: M
authors.lastname: Nishimura
authors.forename: Hana
authors.initials: H
authors.lastname: Takahashi
authors.forename: Chitoshi
authors.initials: C
authors.lastname: Kitamura
authors.forename: Dai-Ichiro
authors.initials: D
authors.lastname: Kato
authors.forename: Seiji
authors.initials: S
authors.lastname: Negoro
mesh_terms: D000478:Alkylation; D000595:Amino Acid Sequence; D001673:Biodegradation, Environmental; D050557:Catechol 2,3-Dioxygenase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation; D004795:Enzyme Stability; D007700:Kinetics; D008969:Molecular Sequence Data; D010636:Phenols / Q000378:metabolism*; D016958:Pseudomonas putida / Q000145:classification* / Q000201:enzymology*; D013045:Species Specificity
chemical_list: D010636:Phenols; D050557:Catechol 2,3-Dioxygenase