International Journal of Microbiology Research, ISSN: 0975-5276, Volume 1, Issue 2, 2009, pp-09-12 Copyright © 2009, Bioinfo Publications, International Journal of Microbiology Research, ISSN: 0975-5276, Volume 1, Issue 2, 2009 Optimization of production and partial purification of laccase by Phanerochaete chrysosporium using submerged fermenation Jhadav A. 1 , Vamsi K.K. 2 , Khairnar Y. 1 , Boraste A. 3 , Gupta N. 5 , Trivedi S. 4 , Patil P. 6 , Gupta G. 7 , Gupta M. 5 , Mujapara A.K. 8 , Joshi B. 9 , Mishra D. 10 1 Padmashree Dr. D.Y. Patil University, Navi Mumbai, 400614, India 2 Rai foundations College CBD Belapur Navi Mumbai 3 S.V.P.M. College of Pharmacy, Baramati, Pune 4 V.V.P. Engineering College, Rajkot, Gujrat 5 Sindhu Mahavidyalaya Panchpaoli Nagpur 6 Dr. D. Y. Patil ACS College, Pimpri, Pune 7 S.D.S.M. College Palghar, Mumbai 8 Sir PP Institute of Science, Bhavnagar, Gujrat, adarshmujapara@gmail.com 9 Rural College of Pharmacy, D.S Road, Bevanahalli, Banglore 10 Senior Research Coordinator, Reliable analytical laboratory, Thane Abstract- Laccases are multi copper oxidases having wide substrate specificity mainly found in white-rot fungi, which are the only microorganisms able to degrade the whole wood components. In contrast to most enzymes, which are generally very substrate specific, laccases act on a surprisingly broad range of substrates, including diphenols, polyphenols, different substituted phenols, diamines, aromatic amines, benzenethiols and even some inorganic compounds such as iodine. As they are capable of degrading a wide variety of compounds they are commercially very significant. This project aims at studying the production optimization of laccase using different carbon sources. Introduction Proteins that contain copper atoms as cofactors are crucial for numerous reactions in cellular metabolism. They are involved in photosynthesis, oxidative phosphorylation, metal ion homeostasis and catabolism of various nutrients and toxic chemical compounds. Laccase is one such copper protein belonging to the oxidoreductase family [1,2]. When oxidized by a laccase, the reducing substrate loses a single electron and usually forms a free radical. The unstable radical may undergo further laccase-catalysed oxidation or non-enzymatic reactions including hydration, disproportionation and polymerization. Figure no.1 shows the schematic laccase-catalyzed oxidation of a p-diphenol. In laccase-mediated reactions, diphenolic compounds undergo a four- electron oxidation. Fig. 1- Laccase-catalyzed oxidation of a diphenol Laccase is used industrially for pulp delignification, polycyclic aromatic hydrogen degradation, pesticide or insecticide degradation and organic synthesis. With microbial enzymes dominating world markets, more innovation and improvisation is needed to increase the efficiency of production at an economical rate [3]. Laccase is the most widely distributed of all the large blue copper-containing proteins, as it is found in a wide range of higher plants and fungi as well as in bacteria. Laccases in plants have been identified in trees, cabbages, turnips, beets, apples, asparagus, potatoes, pears, and various other vegetables. Laccases have been isolated from Ascomyceteous, Deuteromyceteous and Basidiomyceteous fungi. In the fungi, Ascomycetes and Deuteromycetes have not been a focus for lignin degradation studies as much as the white-rot Basidiomycetes. Laccase from Monocillium indicum was the first laccase to be characterized from an ascomycete showing peroxidative activity. The white-rot basidiomycetes are the most efficient degraders of lignin and also the most widely studied. The enzymes implicated in lignin degradation are: lignin peroxidase, which catalyses the oxidation of both phenolic and non-phenolic units, manganese-dependant peroxidase and laccase, which oxidizes phenolic compounds to give phenoxy radicals and quinines; glucose oxidase and glyoxal oxidase for H2O2 production and cellobiose-quinone oxidoreductase for quinone reduction. Phanerochaete chrysosporium is a crust fungus, which forms flat fused reproductive fruiting bodies instead of the mushroom structure. This fungi exhibit an interesting pattern of septate hyphae, giving a stronger line of defense in times of distress. The hyphae network has some branching, with diameters ranging from 3-9 μm. At the ends of the hyphae rests chlamydospores, thick-walled spores varying from 50-60 μm. The conidiophores give rise to round asexual blastoconidia, which are 6-9 μm in diameter. Due to its sustainability at moderate to higher temperatures, specifically 40oC, this white-rot fungus can be found in forests ranging from North America, to areas of Europe and in Iran. Phanerochaete chrysosporium is a saprophytic fungus capable of organic breakdown of the