Concerns over climate change have necessitated a rethinking of our transportation infrastructure. One possible alternative to carbon-polluting fossil fuels is biofuels produced by engineered microorganisms that use a renewable carbon source. Two biofuels, ethanol and biodiesel, have made inroads in displacing petroleum-based fuels, but their uptake has been limited by the amounts that can be used in conventional engines and by their cost. Advanced biofuels that mimic petroleum-based fuels are not limited by the amounts that can be used in existing transportation infrastructure but have had limited uptake due to costs. In this Review, we discuss engineering metabolic pathways to produce advanced biofuels, challenges with substrate and product toxicity with regard to host microorganisms and methods to engineer tolerance, and the use of functional genomics and machine learning approaches to produce advanced biofuels and prospects for reducing their costs.

对气候变化的担忧使我们有必要重新考虑我们的交通基础设施。碳污染化石燃料的一种可能替代品是由使用可再生碳源的工程微生物生产的生物燃料。乙醇和生物柴油这两种生物燃料已在取代石油基燃料方面取得进展，但它们的应用受到传统发动机中可用量及其成本的限制。模仿石油基燃料的先进生物燃料不受现有交通基础设施中可使用的数量的限制，但由于成本而受到限制。在这篇综述中，我们讨论了工程化代谢途径以生产先进的生物燃料、底物和产品毒性对宿主微生物的挑战以及工程耐受性的方法，