This study focused on optimizing endoglucanase production using a peculiar fungal co-culture comprising Rhizopus arrhizus and Aspergillus fumigatus, identified through morphological and 18S rDNA analyses. The co-culture achieved the highest enzyme production after 72 h of fermentation with alkaline-treated substrates. Scanning Electron Microscopy (SEM) revealed substantial structural disruption in pretreated biomass, enhancing enzyme accessibility. Among the tested substrates, pea hulls proved to be the most effective for enzyme production. Optimization of physical and nutritional parameters was performed using Design of Experiments (DOE) approaches, specifically Plackett–Burman Design (PBD) for screening and Central Composite Design (CCD) for fine optimization. The maximum endoglucanase activity of 119.58 U/mL/min was obtained under the optimized conditions of 27.5 °C, pH 5.5, inoculum age 3.5 days, and supplementation with 1.5% fructose, 1.25% yeast extract, 1.25% sodium nitrate, and 1.25% Tween 80. Analysis of Variance (ANOVA) confirmed the significance of these parameters and their interactions at a 95% confidence level, with a strong model fit (R2 = 0.9052). This study demonstrates the potential of waste pea hulls as a cost-effective substrate for enzyme production, supporting waste valorization and contributing to a circular bioeconomy through sustainable biomass utilization.
Ali et al. (Sat,) studied this question.
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