The growing demand for renewable energy has intensified the interest in utilizing agricultural biomass as a sustainable feedstock for bioenergy production. Agricultural residues such as crop waste, forestry by-products, and agro-industrial waste are rich in lignocellulosic components. However, the inherent structural complexity and recalcitrance of these biomaterials pose a major barrier to their efficient conversion into biofuels. Microbial enzymes play a pivotal role in degrading lignocellulosic biomass into fermentable sugars, which facilitates bioenergy generation. The microbial sources of these enzymes range from bacteria and fungi to genetically engineered microorganisms specifically tailored for improved enzyme production and activity. Enzymatic hydrolysis is achieved through the synergistic action of multiple enzymes, which work synergistically to break down complex polymers into fermentable sugars. These sugars are then converted into various bioenergy products, including bioethanol, biogas, biodiesel, biohydrogen, and notably jet biofuel, which is a critical focus in the decarbonization of the aviation industry. Jet biofuel is typically produced via alcohol-to-jet or hydroprocessed esters and fatty acids pathways using intermediates derived from lignocellulosic sugars through enzymatic and microbial processes. Despite considerable technological progress, challenges such as enzyme instability, high production costs, and limited process scalability continue to hinder commercial deployment. To address these issues, ongoing research focuses on enzyme engineering, metabolic optimization of microbial hosts, and bioprocess innovations aimed at improving yield, reducing costs, and ensuring industrial feasibility. This chapter offers a comprehensive overview of microbial enzymes in lignocellulosic biomass conversion, detailing their sources, mechanisms, and roles in sustainable and next-generation biofuel production, including the emerging potential of jet biofuels. It also highlights prevailing challenges and the future prospects of enzymatic technologies in advancing global bioenergy solutions.