One of the highest costs in bio-butanol production is the separation of butanol from the fermentation broth, which is composed of acetone, butanol, and ethanol. Instead of using butanol, researchers experimentally investigated the utilization of the acetone-butanol-ethanol (ABE) mixture in gasoline and diesel engines to promising results. However, there has been no well-to-wheel (WTW) study to determine the life cycle energy balance and greenhouse gas (GHG) emissions that covers the entire production process and use of ABE as a transportation fuel. In this study, the WTW analysis of ABE from two feedstocks (corn and corn stover-based) and their benefits in replacing a portion of gasoline or diesel with ABE is investigated. Results show that ABE could achieve 56-80% fossil fuel savings and 53-83% GHG emission reductions when compared to both gasoline and diesel. The WTW analysis indicated that ABE from both feedstocks has the potential to become as an alternative biofuel directly and as an additive blend for diesel or gasoline. ? 2021 Elsevier Ltd. All rights reserved.