Biocatalytic cascades guide complex, efficient and selective intracellular transformations. These unique features originate from the spatial organization of the biocatalysts in confined cellular environments that allow the directional channelling of reaction intermediates across the cells. Here we address efforts directed towards the development of synthetic cell analogues and supramolecular ensembles acting as nano/microenvironments for operating biocatalytic cascades. Multienzyme systems are integrated within metal–organic frameworks, polymersomes, lipid-stabilized microdroplets and hydrogel microparticles acting as cell-like containments. Also, multienzyme systems are spatially positioned on one-dimensional DNA wires, two-dimensional DNA strips or origami tiles, and three-dimensional DNA origami bundles or cages, and specific protein–protein interactions or peptide–protein complexes provide versatile scaffolds for engineering enzyme assemblies. Biocatalytic cascades operating on these scaffolds or in confined nano/microenvironments reveal substantially enhanced reaction yields compared with the analogous diffusional mixtures of the biocomponents. Mechanistic pathways accounting for the enhanced biocatalytic activities and future challenges in developing and applying biocatalytic cascades are presented.

生物催化级联反应指导复杂，有效和选择性的细胞内转化。这些独特的特征源于在狭窄的细胞环境中生物催化剂的空间组织，该空间组织允许反应中间体在细胞之间定向引导。在这里，我们致力于开发用于操作生物催化级联反应的纳米/微环境的合成细胞类似物和超分子集合体。多酶系统集成在金属-有机框架，聚合物小体，脂质稳定的微滴和充当细胞样容器的水凝胶微粒中。同样，多酶系统在空间上位于一维DNA折线，二维DNA条或折纸砖以及三维DNA折纸束或笼子上，特定的蛋白质-蛋白质相互作用或肽-蛋白质复合物为工程酶组装提供了多功能的支架。与这些生物组分的类似扩散混合物相比，在这些支架上或在受限的纳米/微环境中运行的生物催化级联反应显示出显着提高的反应产率。提出了机制途径，解释了增强的生物催化活性以及在开发和应用生物催化级联反应中的未来挑战。