Enhanced hydrolysis of lignocellulosic biomass with doping of a highly thermostable recombinant laccase.

A highly thermostable laccase from Geobacillus sp. strain WSUCF1 was cloned into Escherichia coli (E. coli) using pRham N-His SUMO expression system. The thermostable laccase with a molecular weight ~30 kDa had a t _1/2 (pH 6.0) of 120 h at 50 °C. The homology modelling for laccase structure showed the presence of Cu active centers with His and Cys residues involved in the active site and ligand binding activity of the enzyme, respectively. The K _m , V _max , K _cat and K _cat /K _m values of the purified enzyme with ABTS were found to be 0.146 mM, 1.52 U/mg, 1037 s ^-1 and 7102.7 s ^-1  mM ^-1 , respectively. The doping of recombinant WSUCF1 laccase to commercial enzyme cocktails Accellerase® 1500 and Cellic CTec2 improved the hydrolysis of untreated, alkali and acid treated corn stover by 1.31-2.28 times and bagasse by 1.32-2.02 times. Further, in-house enzyme cocktails with laccase hydrolyzed untreated, alkali and acid treated bagasse and gave 1.44, 1.1, and 0.92 folds higher sugar, respectively, when compared with Accellerase 1500. The results suggested that thermostable laccase can aid in the improved hydrolysis of lignocellulosic biomass.
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