Characterization of optimized production, purification and application of laccase from Ganoderma lucidum

Abstract

We show for the first-time Ganoderma lucidum laccase enzyme production using medium containing 3% (v/v) ethanol, which enhanced the enzyme production up to 14.1 folds. A more than 400-folds increase could be achieved if grown in the presence of the novel lignocellulosic biomass tamarind shell plus ethanol (3%, v/v), CuSO4 (0.4 mM) and gallic acid (1 mM). A 38.3 kDa laccase enzyme was purified from the initial protein preparation with an overall yield of 32% using Sephadex G-100 and DEAE-cellulose column chromatography. The enzyme was identified through MALDI-TOF/TOF tandem mass spectrometry (MS/MS) as G. lucidum laccase-3. This enzyme exerted its optimal activity at a pH of 5 and a temperature of 55 degrees C with ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) as an ideal substrate. The catalytic efficiencies (k(cat)/K-m) determined for ABTS and guaiacol were 11.5 x 10(5) and 3.9 x 10(5)s(-1) M-1, respectively. The G. lucidum laccase decolorized various textile dyes and industrial textile dye effluent up to 90% and 97%, respectively. (C) 2012 Elsevier B.V. All rights reserved.