~ 1977 ~  Journal of Pharmacognosy and Phytochemistry 2020; 9(1): 1977-1985             E-ISSN: 2278-4136 P-ISSN: 2349-8234 www.phytojournal.com JPP 2020; 9(1): 1977-1985 Received: 23-11-2019 Accepted: 26-12-2019   Sachchidanand Ex P.G scholar, Department of Industrial Microbiology, SHUATS Allahabad, Uttar Pradesh, India Ajay Kumar Singh Department of Food Process Engineering, SHUATS Allahabad, Uttar Pradesh, India Yashab Kumar Department of Industrial Microbiology, SHUATS Allahabad, Uttar Pradesh, India Harison Masih Department of Industrial Microbiology, SHUATS Allahabad, Uttar Pradesh, India Kunal Singh Researcher, Department of Food Process Engineering, SHUATS Allahabad, Uttar Pradesh, India Corresponding Author: Kunal Singh Researcher, Department of Food Process Engineering, SHUATS Allahabad, Uttar Pradesh, India Industrial application of fungal laccase produced by solid state fermentation of agro pulse by products Sachchidanand, Ajay Kumar Singh, Yashab Kumar, Harison Masih and Kunal Singh Abstract Laccase is a copper containing polyphenol oxidase that acts on a wide range of substrate. A laccase producing fungi (Trichoderma spp.) was isolated and subjected to purification, characterization and dye decolorization study. Laccase activity was highest when operated at the following condition, Bengal gram used as a solid state substrate, 7 days incubation, 25 °C temperature, and pH-6 and inoculum size 2%. Guaiacol and sodium acetate buffer were used to assay laccase production. The enzyme was partially purified by ammonium sulphate precipitation and dialysis method. The purified enzyme was obtained with a specific activity of 89.959 IU/mg protein and a final yield of 70.26%. The optimal pH and temperature of the laccase was recorded to be 5 and 45 °C respectively. The values of kinetic parameters Km and Vmax for purified laccase were noted at 0.055 and 37.037 respectively, for guaiacol as substrate. The enzyme activity of crude enzyme and partially purified enzyme was observed to be 18.181 IU/ml and 44.710 IU/ml respectively. 63.866% of decolorization was observed at 48 h with Congo red and 19.009% with Rhodamine B by laccase enzyme produced by Trichoderma spp. is able to decolorize Congo red dye and Rhodamine B and can be used in controlling environmental pollution. Keywords: Laccase, Trichoderma, Guaiacol, Dye decolorization, Partial purification 1. Introduction Laccase is an enzyme that has potential ability of oxidation. It belongs to those enzymes, which have innate properties of reactive radical production, and its utilization in many fields has been ignored because of its unavailability in the commercial field. There are diverse sources of laccase producing organisms like bacteria, fungi and plants. Textile, pulp and paper industries discharge a huge quantity of waste in the environment, and the disposal of this waste is a big problem. To solve this problem, work has done to discover such an enzyme, which can detoxify these wastes and is not harmful to the environment. Laccase use oxygen and produce water as by product. They can degrade a range of compounds including phenolic and non- phenolic compounds. They also have ability to detoxify a range of environmental pollutants. Laccase are typically found in plants and fungi. Plant laccase participate in the radical-based mechanisms of lignin polymer formation [13, 29] whereas in fungi laccase probably have more roles including morphogenesis, fungal plant-pathogen/host interaction and stress defense and lignin degradation. Although there are also some reports about laccase activity in bacteria Solid state fermentation (SSF) is environmental friendly as it resolves the problem of solid wastes disposal. It has been generally claimed that product yields are mostly higher in SSF when compared to submerged fermentation (SmF). Production of these biocatalysts using agro-biotech substrates under solid-state fermentation conditions provide several advantages in productivity, cost-effectiveness in labour, time and medium components in addition to environmental advantages like less effluent production, waste minimization, etc [20] . There are several reports describing use of agro-industrial residues for the production of laccase e.g. Banana skin by Trametes pubescens [19] , Coconut flesh, groundnut shells and groundnut seeds by T. hirsute [4] . Most of these wastes contains lignin or/and cellulose and hemicellulose, which act as inducers of the ligninolytic activities. Moreover, most of them are rich in sugars, which make the whole process much more economical. All these make them very suitable as raw materials for the production of secondary metabolites of industrial significance by microorganisms.