Characterization of a multifunctional enzyme from Trichoderma harzianum and its application in enhanced enzymatic hydrolysis

Efficient saccharification of lignocellulose to fermentable sugars is crucial for bioconversion, yet the process is often hindered by insufficient β-glucosidase, β-xylosidase, and α-L-arabinofuranosidase activities in enzyme cocktails from Trichoderma reesei. This study addresses this gap by identifying BX1, a multifunctional enzyme from the underexplored fungus Trichoderma harzianum EM0925, which demonstrates a triad of activities targeting hemicellulose-derived oligosaccharides preferentially. We used structural analysis, molecular docking, and mutation studies to elucidate the roles of specific residues (Asp