Biofuel production has been promoted as a means of saving fossil fuels and reducing greenhouse gas (GHG) emissions. However, there are concerns about the potential of biofuel to improve energy efficiency and contribute to climate change mitigation. This paper investigates energy efficiency, GHG emission performance, and the cost-effectiveness of biofuel as energy for transportation. Energy and GHG balances are calculated for a functional unit of 1 km using life-cycle assessment and considering the effects of land use change (LUC) and managed soils in feedstock production. Based on study results, the ethanol substitution for gasoline in the form of E5 and E10 would achieve energy savings, provided that their fuel consumption (in terms of l km-1) compared to gasoline does not increase by more than 2.4% and 4.5%, respectively. This substitution achieves a GHG emission saving, provided that the fuel consumption of E5 and E10 compared to gasoline does not increase by more than 3.8% and 7.8%, respectively. The substitution of biodiesel for diesel in the form of B5 and B10 would achieve an energy saving, provided that their fuel consumption compared to diesel does not increase by more than 0.66% and 4.1%, respectively. Compared to biodiesel, the production and use of ethanol demonstrates higher cost-effectiveness, energy efficiency, and acceptable GHG emissions performance.