title: Conversion of 2-fluoromuconate to cis-dienelactone by purified enzymes of Rhodococcus opacus 1cp.
abstract: The present study describes the (19)F nuclear magnetic resonance analysis of the conversion of 3-halocatechols to lactones by purified chlorocatechol 1,2-dioxygenase (ClcA2), chloromuconate cycloisomerase (ClcB2), and chloromuconolactone dehalogenase (ClcF) from Rhodococcus opacus 1cp grown on 2-chlorophenol. The 3-halocatechol substrates were produced from the corresponding 2-halophenols by either phenol hydroxylase from Trichosporon cutaneum or 2-hydroxybiphenyl 3-mono-oxygenase from Pseudomonas azelaica. Several fluoromuconates resulting from intradiol ring cleavage by ClcA2 were identified. ClcB2 converted 2-fluoromuconate to 5-fluoromuconolactone and 2-chloro-4-fluoromuconate to 2-chloro-4-fluoromuconolactone. Especially the cycloisomerization of 2-fluoromuconate is a new observation. ClcF catalyzed the dehalogenation of 5-fluoromuconolactone to cis-dienelactone. The ClcB2 and ClcF-mediated reactions are in line with the recent finding of a second cluster of chlorocatechol catabolic genes in R. opacus 1cp which provides a new route for the microbial dehalogenation of 3-chlorocatechol.
authors.affiliation: G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russian Federation.
authors.forename: Inna P
authors.initials: IP
authors.lastname: Solyanikova
authors.forename: Olga V
authors.initials: OV
authors.lastname: Moiseeva
authors.forename: Sjef
authors.initials: S
authors.lastname: Boeren
authors.forename: Marelle G
authors.initials: MG
authors.lastname: Boersma
authors.forename: Marina P
authors.initials: MP
authors.lastname: Kolomytseva
authors.forename: Jacques
authors.initials: J
authors.lastname: Vervoort
authors.forename: Ivonne M C M
authors.initials: IM
authors.lastname: Rietjens
authors.forename: Ludmila A
authors.initials: LA
authors.lastname: Golovleva
authors.forename: Willem J H
authors.initials: WJ
authors.lastname: van Berkel
mesh_terms: D000272:Adipates / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D049308:Dioxygenases; D019753:Intramolecular Lyases / Q000302:isolation & purification / Q000378:metabolism*; D007783:Lactones / Q000302:isolation & purification / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D010105:Oxygenases / Q000302:isolation & purification / Q000378:metabolism*; D012240:Rhodococcus / Q000201:enzymology
chemical_list: D000272:Adipates; D007783:Lactones; C001240:2-fluoromuconic acid; C084081:dienelactone; D010088:Oxidoreductases; D010105:Oxygenases; D049308:Dioxygenases; C044173:chlorocatechol 1,2-dioxygenase; D019753:Intramolecular Lyases; C055773:chloromuconate cycloisomerase