Reprogramming Nonribosomal Peptide Synthetases for Site-Specific Insertion of α-Hydroxy Acids
Autor: | Camus, Anna, Truong, Gisèle, Mittl, Peer R E, Markert, Greta, Hilvert, Donald |
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Přispěvatelé: | University of Zurich, Hilvert, Donald |
Rok vydání: | 2022 |
Předmět: |
Biological Products
1303 Biochemistry 1503 Catalysis 610 Medicine & health 1600 General Chemistry 1505 Colloid and Surface Chemistry General Chemistry Biochemistry Catalysis Substrate Specificity Colloid and Surface Chemistry Depsipeptides 10019 Department of Biochemistry 570 Life sciences biology Amines Amino Acids Peptide Synthases Hydroxy Acids |
Zdroj: | Journal of the American Chemical Society. 144(38) |
ISSN: | 1520-5126 |
Popis: | High-throughput engineering has the potential to revolutionize the customization of biosynthetic assembly lines for the sustainable production of pharmaceutically relevant natural product analogs. Here, we show that the substrate specificity of gatekeeper adenylation domains of nonribosomal peptide synthetases can be switched from an α-amino acid to an α-hydroxy acid in a single round of combinatorial mutagenesis and selection using yeast cell surface display. In addition to shedding light on how such proteins discriminate between amino and hydroxy groups, the remodeled domains function in a pathway context to produce α-hydroxy acid-containing linear peptides and cyclic depsipeptides with high efficiency. Site-specific replacement of backbone amines with oxygens by an engineered synthetase provides the means to probe and tune the activities of diverse peptide metabolites in a simple and predictable fashion. |
Databáze: | OpenAIRE |
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