Production, purification, characterization and application of a new halotolerant and thermostable endoglucanase of Botrytis ricini URM 5627.

A halotolerant endoglucanase with a molecular mass of 39 kDa was obtained from the solid fermentation of sugarcane bagasse by the fungus Botrytis ricini URM 5627 and isolated using only two purification processes: fractionation with ammonium sulphate and size-exclusion chromatography resulting in an activity of 1289.83 U/mL. After the isolation, biochemical characterizations were performed, giving a temperature of 50 °C and optimum pH of 5. The enzyme was stable at 39-60 °C for 60 min and at a pH of 4-6. The enzymatic activity increased in the presence of Na+, Mn2+, Mg2+ and Zn2+ and decreased in the presence of Ca2+, Cu2+, and Fe2+. The endoglucanase revealed a halotolerant profile since its activity increased proportionally to an increase in NaCl concentration. The maximum activity was reached at 2 M NaCl with a 75% increase in activity. The enzyme had a Km of 0.1299 ±â€¯0.0096 mg/mL and a Vmax of 0.097 ±â€¯0.00121 mol/min/mL. During application in saccharification tests, the enzyme was able to hydrolyse sugarcane bagasse, rice husk, and wheat bran, with the highest production of reducers/fermentable sugars within 24 h of saccharification for wheat bran (137.21 mg/g). Therefore, these properties combined make this isolated enzyme a potential candidate for biotechnological and industrial applications.