Moving cannabinoid production away from the vagaries of plant extraction and into engineered microbes could provide a consistent, purer, cheaper and environmentally benign source of these important therapeutic molecules, but microbial production faces notable challenges. An alternative to microbes and plants is to remove the complexity of cellular systems by employing enzymatic biosynthesis. Here we design and implement a new cell-free system for cannabinoid production with the following features: (1) only low-cost inputs are needed; (2) only 12 enzymes are employed; (3) the system does not require oxygen and (4) we use a nonnatural enzyme system to reduce ATP requirements that is generally applicable to malonyl-CoA-dependent pathways such as polyketide biosynthesis. The system produces ~0.5 g l⁻¹ cannabigerolic acid (CBGA) or cannabigerovarinic acid (CBGVA) from low-cost inputs, nearly two orders of magnitude higher than yeast-based production. Cell-free systems such as this may provide a new route to reliable cannabinoid production.

将大麻素生产从变幻莫测的植物提取转移到工程微生物中可以为这些重要的治疗分子提供一致、更纯净、更便宜和环境友好的来源，但微生物生产面临着显着的挑战。微生物和植物的替代方案是通过采用酶生物合成来消除细胞系统的复杂性。在这里，我们设计并实现了一种用于大麻素生产的新型无细胞系统，具有以下特点：（1）仅需要低成本投入；(2)仅使用12种酶；(3) 该系统不需要氧气，(4) 我们使用非天然酶系统来减少 ATP 需求，这通常适用于丙二酰辅酶 A 依赖性途径，如聚酮化合物生物合成。该系统利用低成本投入生产~0.5 g l ^{-1大麻二酚酸（CBGA）或大麻二酚酸（CBGVA），比基于酵母的生产高出近两个数量级。}诸如此类的无细胞系统可能为可靠的大麻素生产提供一条新途径。