Optimization of Temperature, Cellulase Concentration and pH value of Enzymatic Saccharification for Producing Sugar from Ozone Pre-treated Oil Palm Empty Fruit Bunch

Enzymatic saccharification is a crucial step in biomass conversion, where cellulase enzymes break down cellulose into fermentable sugars. This study focuses on optimizing the enzymatic saccharification process for total reducing sugar (TRS) production from ozone pre-treated oil palm empty fruit bunches (OPEFB). A Face-Centred Central Composite Design (FCCD) was employed to evaluate the effects of three key process parameters: temperature (30 °C to 60 °C), cellulase concentration (0.5 mg/mL to 1.5 mg/mL), and pH (3 to 7). A total of 20 experimental runs were conducted. The response variable measured was the concentration of total reducing sugar (TRS) produced. The highest TRS yield achieved was 3.598 mg/mL under the optimized conditions of 39.157 °C, 1.177 mg/mL cellulase concentration, and pH 6.824. These findings demonstrate that ozone pre-treatment significantly enhances the enzymatic hydrolysis efficiency of OPEFB. The optimized conditions suggest a promising pathway for the efficient production of TRS, supporting future upscaling and potential commercialization of bio-based products.