<p>The global pursuit of sustainable and renewable energy sources has intensified interest in biofuel production from lignocellulosic biomass. There are challenges for achieving sustainable biofuel production and utilization of lignocelluloses through microbial applications. The feedstock selection, pretreatment techniques, enzymatic hydrolysis, fermentation, and purification are important considerations from a sustainability perspective. Non-edible biomass sources, including agricultural and forest residues, are highlighted for their potential to reduce competition with food crops and minimize environmental impacts. Various pretreatment methods are explored for their efficacy in breaking down the complex lignocellulosic structure and enhancing enzymatic accessibility. Advances in enzyme technologies, metabolic engineering, and microbial biotechnology have significantly improved the efficiency of enzymatic hydrolysis and fermentation processes, resulting in increased sugar release and higher biofuel yields. The review emphasizes sustainability aspects, including energy security, reduced greenhouse gas emissions, and the utilization of renewable resources, in the context of microbial applications. However, overcoming technical and economic challenges, scaling up production, and ensuring commercial viability require further research and development. Continual advancements in microbial processes, coupled with innovation and comprehensive sustainability assessments, hold substantial promise for the sustainable production of biofuels from lignocellulosic biomass, contributing to a greener and more resilient energy future.</p> Graphical Abstract <p></p>