Mechanistic Insights into Fractal Kinetics and Cellulase Adsorption in the Saccharification of Avicel PH-101 and Pretreated Hemp Hurd

he enzymatic saccharification of cellulose is governed by heterogeneous reaction environments that deviate from classical Michaelis-Menten behavior. Methods: Fractal kinetics were applied to describe the hydrolysis of microcrystalline cellulose (Avicel PH-101) and pretreated hemp hurds using Cellic CTec2. Optimal enzyme loading was first established on Avicel, and the influence of mixing regimes was evaluated. Results: Rotational agitation markedly improved hydrolysis efficiency. Organosolv-based pretreatments generated cellulose-enriched substrates that exhibited higher reactivity than Avicel, while redeposited lignin showed minimal inhibitory effects. Enzyme adsorption studies revealed substantial binding to lignocellulosic substrates, suggesting nonspecific interactions and crowding effects that influence kinetic parameters. Conclusions: Fractal coefficients k and h successfully captured differences in substrate accessibility and reactivity, demonstrating the suitability of fractal models for describing cellulose saccharification in complex solid-liquid systems. Organosolv pretreatment allows a high degree of saccharification, whereas redeposited lignin does not interfere with the enzymatic reaction.