Purification and characterization of maleylacetate reductase from Nocardioides simplex 3E utilizing phenoxyalcanoic herbicides 2,4-D and 2,4,5-T.
Maleylacetate reductase was isolated and purified from the Gram-positive strain Nocardioides simplex 3E which is able to utilize the phenoxyalcanoic herbicides 2,4-D and 2,4,5-T. Cells were grown on 2,4-D as the sole carbon source. The enzyme was purified by 380-fold with 3.0% yield. The purified maleylacetate reductase is a homodimer with subunit molecular mass of 37 kD. The enzyme required NADH as a cofactor; the Km for maleylacetate is 25 microM; Vmax (with NADH as cofactor) and kcat are 185 U/mg and 6845 min-1, respectively. The enzyme is very unstable; its pH and temperature optima are at 7.0-7.1 and 50 degrees C, respectively.
Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia.
V M
VM
Travkin
E V
EV
Linko
L A
LA
Golovleva
D015085:2,4,5-Trichlorophenoxyacetic Acid / Q000494:pharmacology*; D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology*; D000192:Actinomycetales; D004305:Dose-Response Relationship, Drug; D006540:Herbicides / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009618:Nocardiaceae / Q000201:enzymology*; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification*; D044925:Oxidoreductases Acting on CH-CH Group Donors; D013696:Temperature
D006540:Herbicides; D015084:2,4-Dichlorophenoxyacetic Acid; D015085:2,4,5-Trichlorophenoxyacetic Acid; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C021530:maleylacetate reductase