The term "biofuel" refers to a liquid or gas fuel obtained mostly from biomass for use in transportation. Biobutanol is a potential replacement biofuel for fossil-based liquid fuels as they become depleted. Biobutanol is a transportation fuel that may be simply combined with either gas or petrol at any ratio. Clostridia are the most prevalent fermentative organisms used in biobutanol production. It might well be recognized for its ability to use the acetone-butanol-ethanol (ABE) fermentation route to convert various types of renewable biomass to biobutanol. Various aspects of biobutanol fermentation, including butanol toxicity and product titer, have also been addressed. The recent advancement in lignocellulosic biomass treatment technology, which is significantly greener and safer for the environment, has been clearly articulated. This chapter also covers several metabolic engineering and simple engineering laboratory approaches such as Adaptive Laboratory Evolution (ALE) for strain improvement to overcome butanol toxicity. These biobutanol difficulties could be solved to improve microorganism resistance to high solvent concentrations and hence increase biobutanol output. The final section of this chapter will discuss the future of biobutanol production as a new sustainable and renewable future transportation fuel. Overall, this chapter will provide a better grasp of current trends and issues in biomass-based biobutanol production.