Nanocontainers or macromolecular scaffolds for artificial biocatalytic cascades facilitate sequential enzyme reactions but diffusive escape of intermediates limits rate enhancement. Nonribosomal peptide synthetases (NRPS) naturally form gigantic assembly lines and prevent escape by covalently tethering intermediates. Here, we have built DNA-templated NRPS (DT-NRPS) by adding zinc finger tags to split NRPS modules. The zinc fingers direct the NRPS modules to 9-bp binding sites on a DNA strand, where they form a catalytically active enzyme cascade. DT-NRPS outperform previously reported DNA templated enzyme cascades in terms of DNA acceleration which demonstrates that covalent intermediate channeling is possible along the DNA template. Attachment of assembly line enzymes to a DNA scaffold is a promising catalytic strategy for the sequence-controlled biosynthesis of nonribosomal peptides and other polymers.

中文翻译：

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工程DNA模板化非核糖体肽合成

用于人工生物催化级联反应的纳米容器或大分子支架可促进顺序酶反应，但中间体的扩散逸出限制了速率的提高。非核糖体肽合成酶（NRPS）自然形成巨大的装配线，并通过共价束缚中间体防止逸出。在这里，我们通过添加锌指标签来拆分NRPS模块，从而构建了DNA模板NRPS（DT-NRPS）。锌指将NRPS模块引导至DNA链上的9 bp结合位点，在那里它们形成了催化活性的酶级联反应。DT-NRPS在DNA加速方面优于先前报道的DNA模板化酶级联反应，这表明共价中间通道可能沿着DNA模板发生。