Pass-back chain extension expands multimodular assembly line biosynthesis

TitlePass-back chain extension expands multimodular assembly line biosynthesis
Publication TypeJournal Article
Year of Publication2020
AuthorsZhang J.J, Tang X.Y, Huan T., Ross AC, Moore BS
Date Published2020/01
Type of ArticleArticle
ISBN Number1552-4450
Accession NumberWOS:000503057200012
Keywordsadenylation; Biochemistry & Molecular Biology; condensation; crystal-structure; diversity; domain; gene-cluster; module; nonribosomal peptide synthetases; polyketide synthase; products; proteins

Modular nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) enzymatic assembly lines are large and dynamic protein machines that generally effect a linear sequence of catalytic cycles. Here, we report the heterologous reconstitution and comprehensive characterization of two hybrid NRPS-PKS assembly lines that defy many standard rules of assembly line biosynthesis to generate a large combinatorial library of cyclic lipodepsipeptide protease inhibitors called thalassospiramides. We generate a series of precise domain-inactivating mutations in thalassospiramide assembly lines, and present evidence for an unprecedented biosynthetic model that invokes intermodule substrate activation and tailoring, module skipping and pass-back chain extension, whereby the ability to pass the growing chain back to a preceding module is flexible and substrate driven. Expanding bidirectional intermodule domain interactions could represent a viable mechanism for generating chemical diversity without increasing the size of biosynthetic assembly lines and challenges our understanding of the potential elasticity of multimodular megaenzymes.

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