The metabolic engineering approach for production of bioethanol from second generation biomass (portion of non-feeding purpose) is the most emerging research matter in the field of biotechnology. The second generation bio-mass such as agricultural waste, saw dust, grasses, etc. are mainly composed of cellulose, hemi cellulose, and lignin making biomass resistant to degradation. Lignin is one of the major components of lignocellulosic biomass after cellulose and hemi-cellulose. The hydrophobic nature of lignin prevents hydrolysis of cellulose and hemi-cellulose to release fermentable sugar such as glucose and xylose. The three copper centered metalloezyme, laccase of class oxidoreductase (EC catalyze oxidation of lignin resulting to hydrolysis of hemicellulose and cellulose. The basidiomycetes such as White rot fungi, play an important role in lignin degradation by producing laccase enzyme. The laccase encoding 1578 bp synthetic DNA was cloned under GPD promoter for over production of laccase enzyme in Saccharomyces cerevisiae. The recombinant S. cerevisae would be capable of degrading lignin, thus producing bio-ethanol from the lignocellulosic biomass without pretreatment.

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