Folia Microbiologica

, 45:447

Purification and characterization of laccase-1 fromPleurotus florida

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DOI: 10.1007/BF02817619

Cite this article as:
Das, N., Chakraborty, T.K. & Mukherjee, M. Folia Microbiol (2000) 45: 447. doi:10.1007/BF02817619

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 gelfiltration chromatography, respectively. The pI value of L1 has been determined to be 4.1. The optimum reaction temperature of the enzyme is 50°C. TheKm 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.

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© Folia Microbiologica 2000

Authors and Affiliations

  1. 1.Indian Institute of Chemical BiologyCalcuttaIndia