Laccase is among the major enzymes which plays an important role in ligninolytic system of fungi. Laccase production by Aspergillus heteromorphus was studied using anaerobically treated distillery spent wash (ADSW) and lignocellulosic biomass. Lignocellulosic biomass (rice straw, wheat straw and sugarcane bagasse) generated during biomass processing leads to solid waste and distillery spent wash is unwanted liquid waste produced by distilleries, both causes environmental pollution. Two mineral media and anaerobically treated distillery spent wash medium was tested for laccase production. Enzyme production in various media and in presence and absence of lignocellulosic biomass supplements showed that anaerobically treated distillery spent wash medium was a better laccase inducer medium than the mineral media. Addition of lignocellulosic biomass enhances laccase production and highest laccase activity was obtained in 5% anaerobically treated distillery spent wash medium with rice straw. (C) 2009 Elsevier B.V. All rights reserved.