Optimization and Bioreactor Scale-Up of Cellulase Production in Trichoderma sp. KMF006 for Higher Yield and Performance

This study optimized operating parameters to enhance cellulase production and evaluated scale-up feasibility in submerged fermentation (SmF) using <i>Trichoderma</i> sp. KMF006. Flask-scale experiments assessed the effects of Avicel:cellulose ratios (4:0-0:4), agitation speeds (150-210 rpm), and turbulence (baffled vs. non-baffled flasks), with optimized conditions applied to a 10 L bioreactor. A 3:1 Avicel:cellulose ratio (A3C1) significantly accelerated cellulase production, reaching peak activity 6 days earlier than Avicel alone. An agitation speed of 180 rpm was optimal, balancing enzyme activity and energy efficiency. Turbulence enhanced cellulase yields, with baffled flasks increasing EG, BGL, and CBH activities 19.9-, 6.2-, and 8.9-fold, respectively, compared to the control. Biochar further improved cellulase production but only under turbulent conditions, demonstrating a synergistic effect. At the bioreactor scale, the A3-180_Imp (A3C1, 180 rpm, impeller-induced turbulence) achieved the highest enzymatic activity (33.60 U/mL EG, 3.46 U/mL BGL, and 0.63 U/mL CBH). The filter paper unit (FPU) was 84 FPU/mL, a two-fold increase compared to the control. However, excessive turbulence at 210 rpm reduced enzyme stability, emphasizing the importance of balancing shear stress. These findings provide a systematic framework for optimizing SmF conditions, highlighting the significance of balancing hydrodynamic conditions for efficient cellulase production at an industrial scale.