This study aims to optimize the fermentation conditions of Bacillus sp.01 for the simultaneous production of cellulase and reducing sugars from oil palm empty fruit bunch (OPEFB). Milled OPEFB was used as the sole carbohydrate source in a mineral salt medium to investigate the effects of temperature (30-45 °C), pH (5.0-8.0), and nitrogen sources on the fermentation performance of Bacillus sp.01. The results showed that the highest sugar and cellulase production were achieved at pH 7.0 and 35 °C, yielding 2.52 mg/mL and 0.93 U/mL, respectively, in a 100 mL batch system enriched with 1% milled OPEFB. Regression analysis showed that the optimum sugar and cellulase production conditions were pH 6.73 and 32.5°C. Organic nitrogen sources (beef extract and peptone) showed better performance in promoting sugar and cellulase production than inorganic nitrogen sources (NH4 NO3 and (NH4 )2 SO4 ). These results show the potential of using OPEFB as a substrate for bioconversion by Bacillus sp.01, which could contribute to developing sustainable waste management strategies in the palm oil industry.
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