Biofuels are renewable energy sources derived from biological materials and have gained significant attention as alternatives to fossil fuels due to their potential to reduce greenhouse gas emissions and promote sustainable energy production. In the context of environmental biotechnology, the development of biofuels has progressed through multiple generations, each with distinctive characteristics, feedstocks, and technological innovations. The first generation of biofuels primarily relies on food crops that are rich in sugars, starch, or oils. Common examples include sugarcane, maize, wheat, and oilseeds like soybean and rapeseed. The main biofuels produced in this generation are bioethanol and biodiesel. Bioethanol is typically generated through fermentation of sugars extracted from crops like sugarcane or starch from maize, which is enzymatically converted to glucose and subsequently fermented by microorganisms such as *Saccharomyces cerevisiae*. Biodiesel, on the other hand, is produced by transesterification of vegetable oils or animal fats using catalysts like sodium or potassium hydroxide, resulting in fatty acid methyl esters (FAME). The salient features of first-generation biofuels include their relatively simple production technology, well-established industrial infrastructure, and compatibility with existing internal combustion engines. However, the primary drawbacks are significant competition with food resources, large land and water requirements, and environmental impacts such as deforestation and loss of biodiversity. Despite these limitations, first-generation biofuels served as the foundation for bioenergy development, providing proof-of-concept and commercial viability. The second generation of biofuels was developed to overcome the limitations of the first generation by utilizing non-food biomass, such as lignocellulosic materials. These include agricultural residues (e.g., wheat straw, rice husk, corn stover), forestry residues, and energy crops like switchgrass and miscanthus. Second-generation biofuels still produce bioethanol and biodiesel, but the raw materials are primarily composed of cellulose, hemicellulose, and lignin, which are more complex and resistant to degradation. The production involves pretreatment processes to break down lignocellulose into fermentable sugars, often using chemical, physical, or enzymatic methods. Enzymatic