title: Purification and characterization of dichloromuconate cycloisomerase from Alcaligenes eutrophus JMP 134.
abstract: Dichloromuconate cycloisomerase from Alcaligenes eutrophus JMP 134 was purified to homogeneity. The enzyme has an Mr of about 270,000 as determined by gel filtration and consists of six to eight subunits of identical Mr 40,000 as determined by SDS/PAGE. Mn2+ ions as well as thiol groups are required for activity. A high Km value of about 4 mM for cis,cis-muconate explains the reported low activity with this compound. Relatively high Km values were also calculated for monochloro-substituted cis,cis-muconates (300-500 microM), in contrast with the low Km value of 20 microM for 2,4-dichloro-cis,cis-muconate. The catalytic constant of the pure enzyme was 3820 min-1 when measured with 2,4-dichloro-cis,cis-muconate.
authors.affiliation: Institut für Mikrobiologie, Universität Stuttgart, Federal Republic of Germany.
authors.forename: A E
authors.initials: AE
authors.lastname: Kuhm
authors.forename: M
authors.initials: M
authors.lastname: Schlömann
authors.forename: H J
authors.initials: HJ
authors.lastname: Knackmuss
authors.forename: D H
authors.initials: DH
authors.lastname: Pieper
mesh_terms: D000421:Alcaligenes / Q000201:enzymology*; D019753:Intramolecular Lyases; D007535:Isomerases / Q000032:analysis*; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity
chemical_list: D007535:Isomerases; D019753:Intramolecular Lyases; C063594:dichloromuconate cycloisomerase