Abstract
Present study reports a novel and robust GH9 processive endoglucanase β-1,4-glucanase from Bacillus sp. PCH94 (EGase(BL)) with thermo-alkali stable properties. The EGase(BL) gene was cloned in pET-28b(+) and expressed in Escherichia coli BL21(DE3) cells. The recombinant protein was purified 94-fold with a yield of 67.8%. The biochemical characterization revealed an active enzyme at a wide pH (4.0-10.0) and temperature (4-100°C). It showed a Km and Vmax of 1.10 mg/ml and 208.24 IU/mg, respectively, using β-glucan as a substrate. The EGase(BL) showed dual activities for endoglucanase (134.17 IU/mg) and exoglucanase (28.76 IU/mg), assayed using substrates β-glucan and Avicel, respectively. The enzyme is highly stable in neutral and alkaline pH and showed a half-life of 11.29 h, and 8.31 h in pH 7.0 and 9.0, respectively. The enzyme is also compatible with commercial detergents (Tide, Surf, Ghadi, Raj, and Healing tree) of the Indian market and retained > 85% enzyme activity. Concisely, robustness, extreme functionality, and detergent compatibility endorse EGase(BL) as a potential bioresource for the detergent industry, in addition to its implications for the bioethanol industry. Highlights - Cloning, expression, and purification of putative novel GH9 family β-1,4-glucanase.- Processive endoglucanase with CBM3 domain and bi-functional (endo/exo) activity.- Broad pH-temperature active and stable enzyme.- Compatible with commercial detergent powders.