Petro-fuels-driven transportation sector (road, air, and water) is the largest emitter of greenhouse gases (GHGs), several other toxic gases and particulate matters causing rapid deterioration of the atmosphere. Displacing the major transportation fuels namely petrol/gasoline and diesel with suitable biofuels is viewed as the best alternative to bring down the emission/pollution index without imposing restrictions in the mobility of vehicles for communication, business and travel. The most promising alternative of liquid petro-fuels are alcohols, specifically ethanol, which has good fuel properties and compatibility as a blend-in liquid fuel for the existing designs of the internal combustion (IC) engines. While multiple routes of production of ethanol exist, the biochemical production processes are gaining increased popularity due to their potential waste-to-energy aspects and eco-friendly attributes. A few well-known microorganisms are dedicated as the industrial producers of ethanol which convert the fermentable carbohydrates to ethanol through specific solventogenic metabolism. Both edible carbohydrates and non-edible complex biomass can act as potential feedstocks for ethanol production, but the later ones are more favourable as it does not perturb food security. However, the margin of productivity and yield of the ethanol fermentation processes are limited by several factors and need to be elevated to secure its potential as the alternative fuel in the transportation sectors. This chapter presents the current state of the biochemical ethanol production processes and strategies for the improvement to facilitate its implementation as a dependable transportation fuel.