Cellulases are an important industrial enzyme used for bioethanol production. The present research was designed to generate hypercellulase from a locally isolated strain of fungus. For this purpose, fungal strains were collected from local soil, vegetable waste, and agrowaste. The isolated strains were first morphologically identified, then genetically sequenced for further use. The highest cellulase production was achieved by optimizing physico-chemical parameters through RSM by SSF. Furthermore, cellulase was purified using (NH₄)₂SO₄ precipitation, dialysis, and chromatography. The results showed GU227345.1 Aspergillus terreus gave the highest yield (86.665 IU/min/mL) by maintaining parameters, i.e., incubation time of 4 days, temperature of (35ºC), substrate (6 g), pH (6), moisture (90%), inoculum (5 mL), (NH4)2SO4 (0.5 g), and MgSO₄ (0.25 g). It was further enhanced by (NH₄)₂SO₄ precipitation, membrane dialysis, and chromatography. The Km of cellulase was 7.375 mg/mL, and the Vmax value was 1250 mmol min^-1 mg^-1.

Keywords:

Enzyme, Biowaste, Bioethanol, Cellulase, Fungal, SSF, RSM.