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Purification and characterization of laccase-1 fromPleurotus florida

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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. TheK m 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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  1. Indian Institute of Chemical Biology, 700 032, Calcutta, India

    N. Das, T. K. Chakraborty & M. Mukherjee

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  1. N. Das
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  2. T. K. Chakraborty
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  3. M. Mukherjee
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Correspondence to M. Mukherjee.

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Das, N., Chakraborty, T.K. & Mukherjee, M. Purification and characterization of laccase-1 fromPleurotus florida . Folia Microbiol 45, 447–451 (2000). https://doi.org/10.1007/BF02817619

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