Augmenting the Laccase Production in the E.coli Rosetta 2 strain for Future Biotechnological Applications

Laccases have been used for the decolorization and detoxification of synthetic dyes due to their ability to oxidize a wide variety of dyes with water as the sole byproduct. A laccase encoding gene from Bacillus licheniformis was cloned and expressed in E.coli Rosetta2 strain under the control of T7 promoter. Rosetta host strains are Tuner derivatives designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E. coli. To improve the expression the optimal conditions were designed for shake flask experiments. Laccase gene was expressed at 65KDa mol wt size. The ideal condition for laccase production was induction at 0.1mM IPTG, 0.5mM CuCl₂ & incubation at 25°C for 4hrs at 120rpm and then static for overnight to create microaerobic condition. Hence the present study suggests that E.coli Rosetta 2 strain can efficiently express Bacillus licheniformis laccase gene and can be utilized for commercial purpose in future.