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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 (0.4mM) and gallic acid (1mM). A 38.3kDa 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°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 11.5×10⁵ and 3.9×10⁵s …