title: Purification and characterization of laccase-1 from Pleurotus florida.
abstract: Pleurotus florida (ITCC 3308) produces two laccase enzymes (L1 and L2) in potato-dextrose media containing 0.5% yeast extract. Concentrated culture filtrate was separated on DEAE-Sephadex (A-50) column into two enzyme peaks, subsequently named L1 and L2. The L1 enzyme has been purified to homogeneity by ion-exchange and gel-permeation chromatography. L1 is a monomeric glycoprotein with a molar mass of 77 and 82 kDa as determined by SDS-PAGE and gel-filtration chromatography, respectively. The pI value of L1 has been determined to be 4.1. The optimum reaction temperature of the enzyme is 50 degrees C. The Km and some other kinetic parameters of L1 have been determined. Cyanide and azide completely inhibit the enzyme activity. The enzyme was fully active in 1:1 (V/V) buffer-chloroform for at least 2 h. Spectroscopic analysis revealed that the enzyme has four copper atoms, a type 1 copper, a type 2 copper and a type 3 binuclear copper.
authors.affiliation: Indian Institute of Chemical Biology, Calcutta 700 032, India.
authors.forename: N
authors.initials: N
authors.lastname: Das
authors.forename: T K
authors.initials: TK
authors.lastname: Chakraborty
authors.forename: M
authors.initials: M
authors.lastname: Mukherjee
mesh_terms: D003300:Copper / Q000032:analysis; D003470:Culture Media; D007700:Kinetics; D042845:Laccase; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D020076:Pleurotus / Q000201:enzymology* / Q000254:growth & development
chemical_list: D003470:Culture Media; D003300:Copper; D010088:Oxidoreductases; D042845:Laccase