We have developed a system for producing a supramolecular scaffold that permeates the entire Escherichia coli cytoplasm. This cytoscaffold is constructed from a three-component system comprising a bacterial microcompartment shell protein and two complementary de novo coiled-coil peptides. We show that other proteins can be targeted to this intracellular filamentous arrangement. Specifically, the enzymes pyruvate decarboxylase and alcohol dehydrogenase have been directed to the filaments, leading to enhanced ethanol production in these engineered bacterial cells compared to those that do not produce the scaffold. This is consistent with improved metabolic efficiency through enzyme colocation. Finally, the shell-protein scaffold can be directed to the inner membrane of the cell, demonstrating how synthetic cellular organization can be coupled with spatial optimization through in-cell protein design. The cytoscaffold has potential in the development of next-generation cell factories, wherein it could be used to organize enzyme pathways and metabolite transporters to enhance metabolic flux.

我们已经开发出了一种生产能渗入整个大肠杆菌细胞质的超分子支架的系统。该细胞支架由三组分系统构成，该系统包含细菌微区隔壳蛋白和两个互补的从头卷曲螺旋肽。我们表明，其他蛋白质可以靶向这种细胞内的丝状排列。具体地，丙酮酸脱羧酶和醇脱氢酶已被定向到细丝，与不产生支架的那些相比，导致在这些工程细菌细胞中乙醇产生的增加。这与通过酶共配提高的代谢效率相一致。最后，可以将壳蛋白支架直接导向细胞的内膜，这说明如何通过细胞内蛋白设计将合成的细胞组织与空间优化结合在一起。该细胞支架在下一代细胞工厂的发展中具有潜力，其中它可用于组织酶途径和代谢物转运蛋白以增强代谢通量。