Mechanistic analysis of lignocellulosic biomass saccharification by the filamentous fungus Talaromyces cellulolyticus.

Lignocellulosic biomass is a carbon-neutral resource crucial to advancing a bio-based economy. The filamentous fungus Talaromyces cellulolyticus demonstrates superior biomass saccharification efficiency compared to conventional enzyme-producing fungi, making it a promising host for enzymatic biomass conversion. To enable molecular studies, we developed a robust genetic transformation system for T. cellulolyticus and identified key transcription factors regulating saccharifying enzyme genes, classified into three functional groups. Manipulation of these regulators significantly enhanced enzyme production. Additionally, we purified individual enzymes and conducted biochemical and structural analyses, leading to the discovery of a novel xylanase with unique side-chain recognition. The integration of genetic and enzymatic insights advances both our understanding of fungal saccharification systems and the development of more efficient biomass-to-sugar conversion strategies for sustainable biotechnological applications.