Engineered Biosynthesis of Alkyne-Tagged Polyketides by Type I PKSs

• Published in: iScience Vol. 23; no. 3; p. 100938
• Main Authors: Porterfield, William B., Poenateetai, Nannalin, Zhang, Wenjun
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 27.03.2020; Elsevier
• Subjects: BASIC BIOLOGICAL SCIENCES; Biochemical Engineering; Biotechnology; Chemical Engineering; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Metabolic Engineering; Metabolic Engineering; Biotechnology; Chemical Engineering; Biochemical Engineering
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2020.100938

Polyketides produced by modular polyketide synthases (PKSs) are important small molecules widely used as drugs, pesticides, and biological probes. Tagging these polyketides with a clickable functionality enables the visualization, diversification, and mode of action study through bio-orthogonal chemistry. We report the de novo biosynthesis of alkyne-tagged polyketides by modular type I PKSs through starter unit engineering. Specifically, we use JamABC, a terminal alkyne biosynthetic machinery from the jamaicamide B biosynthetic pathway, in combination with representative modular PKSs. We demonstrate that JamABC works as a trans loading system for engineered type I PKSs to produce alkyne-tagged polyketides. In addition, the production efficiency can be improved by enhancing the interactions between the carrier protein (JamC) and PKSs using docking domains and site-directed mutagenesis of JamC. This work thus provides engineering guidelines and strategies that are applicable to additional modular type I PKSs to produce targeted alkyne-tagged metabolites for chemical and biological applications. [Display omitted] •Alkyne-tagged polyketides are de novo biosynthesized using type I PKSs•Docking domains and ACP mutagenesis improve alkyne starter unit translocation•Docking domains, but not ACP mutagenesis, perturb alkyne biosynthetic machinery Chemical Engineering; Biochemical Engineering; Metabolic Engineering; Biotechnology