Fermentation and chemical methods for industrial cytidine diphosphate choline (CDP-choline) catalytic production both suffer from several disadvantages such as relatively low efficiency and productivity. To overcome these problems, we applied the concept of synthetic biology to develop a new one-pot multi-enzyme system to produce CDP-choline from orotic acid. Enzymes from different sources were selected and optimized as building blocks of the system, and parameters such as oxygen supply were also optimized. This system shows a titer of 37.6 ± 1.1 mM and a reaction rate of 1.6 mM L^-1 h⁻¹, both increase 66 % from traditional processes. It also has an efficiency of energy of 25.4%, improves 2-folds. This new one-pot CDP-choline-producing system has a potential for industrial use, and the procedure to design one-pot multi-enzyme system can be applied to build other one-pot system producing energy-rich compounds.

用于工业胞苷二磷酸胆碱 (CDP-胆碱) 催化生产的发酵和化学方法都存在一些缺点，例如效率和生产率相对较低。为了克服这些问题，我们应用合成生物学的概念开发了一种新的单锅多酶系统，以从乳清酸生产 CDP-胆碱。选择和优化来自不同来源的酶作为系统的构建块，并且还优化了供氧等参数。该系统的滴度为 37.6 ± 1.1 mM，反应速率为 1.6 mM L ^-1 h ^-1，两者都比传统工艺增加了 66%。它还具有 25.4% 的能量效率，提高了 2 倍。这种新的一锅 CDP 胆碱生产系统具有工业应用的潜力，设计一锅多酶系统的程序可用于构建其他生产富含能量的化合物的一锅系统。