Two novel routes for the production of gasoline from pyrolysis oil (from timber pine) and biogas (from ley grass) are simulated, followed by a cradle-to-gate life-cycle assessment of the two production routes. The main aim of this work is to conduct a holistic evaluation of the proposed routes and benchmark them against the conventional route of producing gasoline from natural gas. A previously commercialized method of synthesizing gasoline involves conversion of natural gas to syngas, which is further converted to methanol, and then as a last step, the methanol is converted to gasoline. In the new proposed routes, the syngas production step is different; syngas is produced from a mixture of pyrolysis oil and biogas in the following two ways: (i) autothermal reforming of pyrolysis oil and biogas, in which there are two reactions in one reactor (ATR) and (ii) steam reforming of pyrolysis oil and catalytic partial oxidation of biogas, in which there are separated but thermally coupled reactions and reactors (CR). The other two steps to produce methanol from syngas, and gasoline from methanol, remain the same. The purpose of this simulation is to have an ex-ante comparison of the performance of the new routes against a reference, in terms of energy and sustainability. Thus, at this stage of simulations, nonrigorous, equilibrium-based models have been used for reactors, which will give the best case conversions for each step. For the conventional production route, conversion and yield data available in the literature have been used, wherever available.The results of the process design showed that the second method (separate, but thermally coupled reforming) has a carbon efficiency of 0.53, compared to the conventional route (0.48), as well as the first route (0.40). The life-cycle assessment results revealed that the newly proposed processes have a clear advantage over the conventional process in some categories, particularly the global warming potential and primary energy demand; but there are also some in which the conventional route fares better, such as the human toxicity potential and the categories related to land-use change such as biotic production potential and the groundwater resistance indicator. The results confirmed that even though using biomass such as timber pine as raw material does result in reduced greenhouse gas emissions, the activities associated with biomass, such as cultivation and harvesting, contribute to the environmental footprint, particularly the land use change categories. This gives an impetus to investigate the potential of agricultural, forest, or even food waste, which would be likely to have a substantially lower impact on the environment. Moreover, it could be seen that the source of electricity used in the process has a major impact on the environmental performance.