Bioethanol and plant oil-derived biodiesel are generally considered first generation biofuels. Recognizing their apparent disadvantages, scientists and engineers are developing more sustainable and economically feasible second generation biofuels. The new microbial fuels summarized here have great potential to become viable replacements or at least supplements of petroleum-derived liquid transportation fuels. Yields and efficiencies of the four metabolic pathways leading to these microbial fuels-mostly designed and optimized in Escherichia coli and Saccharomyces cerevisiae using modern tools of metabolic engineering and synthetic biology-and the robustness of the biocatalysts that convert the metabolic intermediates to, in some cases, finished and engine-ready fuels, will determine if they can be commercially successful and contribute to alleviating our dependence on fossil fuels.