The past decade has witnessed rapid advance in microbial production of ?drop-in? biofuels from renewable resources. Various biosynthetic pathways have been constructed to produce biofuels with diverse structures, and multiple metabolic engineering strategies have been developed to increase biofuel titers, yields, productivities and system robustness. In this review, we intend to give a brief but comprehensive overview of the most recent progresses on four essential pathways leading to ?drop-in? biofuel production, with an emphasis on the metabolic pathway efficiencies and biofuel structures. Furthermore, we also provide an insightful discussion on optimization strategies to improve the robustness of the microbial platforms for biofuel production.