Molecular Cloning of Laccase Enzyme Producing Gene from Fungal Isolate and Its Expression in E Coli for Industrial Applications

Abstract

Laccases, belonging to the multi-copper oxidase (MCO) family both the newlinehazardous and benign substrates are oxidized by laccase. Applications for it can be newlinefound in the chemical, pharmaceutical, textile, food, and wood processing industries. newlineThis enzyme is an effective catalyst, highly selective, and environmentally friendly as newlineit releases water as its by-product. Large-scale laccase production is crucial for meeting newlinethe demand across various applications. Heterologous production involves expressing newlinelaccase genes in different hosts to enhance output, aiding in isoenzyme identification newlineand minimizing hazardous by-products. Growth parameters significantly influence newlinelaccase production. newlineGrowth parameters significantly impact laccase production, with nutrients such newlineas carbon, nitrogen, and copper being essential. Their concentrations affect enzyme newlinesynthesis differently in various fungi. Agricultural waste, like orange peels and bagasse, newlineis effective and cost-efficient as substrates. Additives such as p-coumaric acid, 2,5- newlinexylidine, and benzoin oxime can enhance laccase production. Operational conditions newlinelike temperature, pH, and aeration levels are crucial. Submerged cultivation in liquid newlinemedia and solid-state fermentation on substrates like lignocellulosic wastes are newlinecommon methods for laccase production. newlineIn the present study, we have isolated Coriolopsis caperata strain and its newlinecapacity to produce laccase is investigated in comparison with other two known laccase newlineproducing organisms Trametes versicolor and Pleurotus ostreatus. Fungal organisms newlinewere screened for laccase production by placing fungal discs on PDA plates containing newlineBromophenol blue. Laccase production was estimated using enzymatic assays, with newlineabsorbance changes indicating enzyme activity.Production involved distinct newlineprocedures for each of the three fungi. To get optimal yield of enzyme from Coriolopsis newlinecaperata Solid-state fermentation was conducted using wheat bran and specific mineral newlinesolutions. Inducers Xylidine and Biotin were added after two days.