Self-sustained metabolic pathways in microcompartments are the corner-stone for living systems. From a technological viewpoint, such pathways are a mandatory prerequisite for the reliable design of artificial cells functioning out-of-equilibrium. Here we develop a microfluidic platform for the miniaturization and analysis of metabolic pathways in man-made microcompartments formed of water-in-oil droplets. In a modular approach, we integrate in the microcompartments a nicotinamide adenine dinucleotide (NAD)-dependent enzymatic reaction and a NAD-regeneration module as a minimal metabolism. We show that the microcompartments sustain a metabolically active state until the substrate is fully consumed. Reversibly, the external addition of the substrate reboots the metabolic activity of the microcompartments back to an active state. We therefore control the metabolic state of thousands of independent monodisperse microcompartments, a step of relevance for the construction of large populations of metabolically active artificial cells.

中文翻译：

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用于代谢功能自下而上整合的失衡微区室。

微隔间中的自我维持代谢途径是生命系统的基石。从技术角度来看，这些途径是可靠设计失衡的人工细胞的必要先决条件。在这里，我们开发了一个微流体平台，用于小型化和分析由油包水液滴形成的人造微隔室中的代谢途径。在模块化方法中，我们将烟酰胺腺嘌呤二核苷酸 (NAD) 依赖性酶促反应和 NAD 再生模块集成到微隔室中作为最小代谢。我们表明，微隔间保持代谢活跃状态，直到底物被完全消耗。可逆地，底物的外部添加将微区室的代谢活动重新启动回活跃状态。