Screening for cellulases and preliminary optimisation of glucose tolerant β-glucosidase production and characterisation.

The search for a novel microbial producer of cellulases including a glucose tolerant β-glucosidase is a challenge as most are inhibited by their product glucose. This study aims to screen for cellulolytic fungi using qualitative and quantitative screening methods. Primary screening revealed 34 of 46 fungal isolates with β-glucosidase activity. Eleven and 13 of these also displayed endoglucanase and exoglucanase activities, respectively. During secondary screening, this number was reduced to 26 β-glucosidase producers with 13 also having endoglucanase and exoglucanase activities. Isolate C1 displayed enhanced production of β-glucosidases in the presence of 0.05 M glucose (69% higher activity). Optimisation of growth conditions for β-glucosidase production by one variable at a time experiments improved production for (isolates) PS1 (64%), MB5 (84%), and C2 (69%). Isolate PS1 identified as sp. BBA70074 displayed the highest tolerance to glucose, retaining 10% of β-glucosidase activity in the presence of 0.8 M glucose. Tolerance to glucose increased to 14% when produced under optimal conditions. β-Glucosidase had a molecular weight of 170 kDa with a pH and temperature optima of 6 and 70°C, respectively. Future studies will include optimisation of the production of the glucose tolerant enzyme by sp. BBA70074.