Chemistry Department, University of Bari Aldo Moro, 70125 Bari, Italy;
Chemistry Department, University of Bari Aldo Moro, 70125 Bari, Italy.
Proc Natl Acad Sci U S A. 2021 Feb 16;118(7). doi: 10.1073/pnas.2012170118.
The construction of energetically autonomous artificial protocells is one of the most ambitious goals in bottom-up synthetic biology. Here, we show an efficient manner to build adenosine 5'-triphosphate (ATP) synthesizing hybrid multicompartment protocells. Bacterial chromatophores from accomplish the photophosphorylation of adenosine 5'-diphosphate (ADP) to ATP, functioning as nanosized photosynthetic organellae when encapsulated inside artificial giant phospholipid vesicles (ATP production rate up to ∼100 ATP∙s per ATP synthase). The chromatophore morphology and the orientation of the photophosphorylation proteins were characterized by cryo-electron microscopy (cryo-EM) and time-resolved spectroscopy. The freshly synthesized ATP has been employed for sustaining the transcription of a DNA gene, following the RNA biosynthesis inside individual vesicles by confocal microscopy. The hybrid multicompartment approach here proposed is very promising for the construction of full-fledged artificial protocells because it relies on easy-to-obtain and ready-to-use chromatophores, paving the way for artificial simplified-autotroph protocells (ASAPs).
构建具有能量自主的人工原细胞是自下而上合成生物学中最具野心的目标之一。在这里，我们展示了一种高效的方法来构建合成的含三磷酸腺苷（ATP）的混合多室原细胞。来自 的细菌类囊体能够将腺苷 5'-二磷酸（ADP）光磷酸化为 ATP，当它们被包裹在人工巨大的磷脂囊泡内时，就可以作为纳米级的光合细胞器发挥作用（每个 ATP 合酶的 ATP 生成速率高达约 100 ATP·s）。通过冷冻电镜（cryo-EM）和时间分辨光谱对类囊体形态和光磷酸化蛋白的取向进行了表征。通过共聚焦显微镜观察到，新合成的 ATP 被用于维持单个囊泡内的 DNA 基因转录，紧随其后的是 RNA 生物合成。这里提出的混合多室方法对于构建成熟的人工原细胞非常有前景，因为它依赖于易于获得和现成的类囊体，为人工简化自养原细胞（ASAPs）铺平了道路。
