Biological CO₂ fixation is so far the most effective means for CO₂ reduction at scale and accounts for most of the CO₂ fixed on Earth. Through this process, carbon is fixed in cellular components and biomass during organismal growth. To uncouple CO₂ fixation from growth and cellular regulation, cell-free CO₂ fixation systems represent an alternative approach since the rate can be independently manipulated. Here we designed an oxygen-insensitive, self-replenishing CO₂ fixation system with opto-sensing. The system comprises a synthetic reductive glyoxylate and pyruvate synthesis (rGPS) cycle and the malyl-CoA-glycerate (MCG) pathway to produce acetyl-coenzyme A (CoA), pyruvate and malate from CO₂, which are also intermediates in the cycle. We solved various problems associated with the in vitro system, and implemented opto-sensing modules to control the regeneration of cofactors. We accomplished sustained operation for 6 hours with a CO₂-fixing rate comparable to or greater than typical CO₂ fixation rates of photosynthetic or lithoautotrophic organisms.

迄今为止，生物固定CO _{2是大规模减少CO 2}的最有效手段，并且占地球上固定CO _{2的大部分。}通过这个过程，碳在有机体生长过程中固定在细胞成分和生物量中。为了将 CO ₂固定与生长和细胞调节分离，无细胞 CO ₂固定系统代表了一种替代方法，因为可以独立控制速率。在这里，我们设计了一种对氧不敏感、自我补充的 CO ₂带光感应的固定系统。该系统包括合成还原乙醛酸和丙酮酸合成 (rGPS) 循环和苹果酰-CoA-甘油酸 (MCG) 途径以从 CO ₂生产乙酰辅酶 A (CoA)、丙酮酸和苹果酸，它们也是循环中的中间体。我们解决了与体外系统相关的各种问题，并实施了光传感模块来控制辅助因子的再生。我们完成了 6 小时的持续操作，其CO ₂固定率与光合或岩石自养生物的典型 CO 2 固定_率相当或更高。