Biomass pretreatment plays a crucial role in the conversion of lignocellulosic biowaste materials into valuable biofuels and biochemicals. Enzymatic pretreatment, in particular, has gained significant attention due to its eco-friendly nature and efficiency in breaking down complex biomass structures. This comprehensive review aims to provide an overview of enzymes used in biomass pretreatment, including cellulases, hemicellulases, ligninases, and their applications in enhancing the efficiency of biomass conversion processes. The review also discusses recent advancements, challenges, and future prospects in the field of enzymatic biomass pretreatment.