Acetyl coenzyme A (CoA) carboxylation is the natural route for endogenous malonyl-CoA formation; however, this pathway presents slow kinetics, carbon and energy inefficiencies, tight regulations and a complicated architecture. These shortcomings limit flux towards malonyl-CoA and become a bottleneck towards the biosynthesis of malonyl-CoA-derived products (MDPs). Here, we design the non-carboxylative malonyl-CoA pathway as a non-natural route for malonyl-CoA biosynthesis, independent from acetyl-CoA. The designed pathway features enzymes such as β-alanine-pyruvate transaminase and malonyl-CoA reductase, exhibits fast kinetics and circumvents tight regulations and the architecture associated with the natural pathway. Furthermore, introducing this pathway into microbes enhances the production of MDPs, including short-chain fatty acids and representative phenol, quinone, alkene, aminoglycoside and macrolide polyketide families, such as spinosad. In summary, this malonyl-CoA formation pathway avoids intrinsic inefficiencies of the natural pathway and can serve as a versatile platform for obtaining MDPs that could be used as fuels, fine chemicals and pharmaceuticals.

乙酰辅酶 A (CoA) 羧化是内源性丙二酰辅酶 A 形成的天然途径；然而，这条途径存在动力学缓慢、碳和能源效率低下、严格的法规和复杂的架构。这些缺点限制了丙二酰辅酶A的通量，并成为丙二酰辅酶A衍生产物（MDP）生物合成的瓶颈。在这里，我们设计了非羧化丙二酰辅酶A途径作为丙二酰辅酶A生物合成的非天然途径，独立于乙酰辅酶A。设计的途径以β-丙氨酸-丙酮酸转氨酶和丙二酰辅酶A还原酶等酶为特征，表现出快速动力学，并规避了严格的法规和与自然途径相关的结构。此外，将该途径引入微生物中可以增强MDP的产生，包括短链脂肪酸和代表性的苯酚、醌、烯烃、氨基糖苷和大环内酯聚酮化合物家族，例如多杀菌素。总之，这种丙二酰辅酶A形成途径避免了自然途径固有的低效率，并且可以作为获得可用作燃料、精细化学品和药品的MDP的多功能平台。