Optimization of Laccase Production by Bacillus sp. Strain AKRC01 in Presence of Agro-waste as Effective Substrate using Response Surface Methodology

Laccases are the widely distributed oxidative enzymes belong to the group of phenoloxidases. These are considered as model enzymes and widely distributed in nature. Bacterial laccases have extremophilic nature and several other unique properties that are not features of fungal laccase such as high-temperature stability and pH. The present study focused on the optimization of laccase production from Bacillus sp. AKRC01 under submerged fermentation state using agro-waste as the substrate. The optimized environmental conditions such as incubation period 96 hours, temperature 35 degrees C and pH 8.0 were obtained maximum enzyme production 3.832 U/ml. In addition, optimized the carbon and nitrogen sources such as 2.0% glucose (4.967 U/mL) and 1.0% peptone (6.236 U/mL) resulted in maximum enzyme production with rice bran as an effective agro-waste substrate. Laccase enzyme was partially purified via ammonium sulfate precipitation method and chromatography using the sephadex column and determination of the molecular weight of 61 kDa compared with the protein ladder was done by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The findings of this study demonstrated the ability of Bacillus sp. AKRC01 using low-cost agro-waste for laccase production. Laccases are important enzymes that have several industrial applications such as bioremediation, bio pulping and food industry.