title: Substrate Flexibility of a Mutated Acyltransferase Domain and Implications for Polyketide Biosynthesis.
abstract: Polyketides are natural products frequently used for the treatment of various diseases, but their structural complexity hinders efficient derivatization. In this context, we recently introduced enzyme-directed mutasynthesis to incorporate non-native extender units into the biosynthesis of erythromycin. Modeling and mutagenesis studies led to the discovery of a variant of an acyltransferase domain in the erythromycin polyketide synthase capable of accepting a propargylated substrate. Here, we extend molecular rationalization of enzyme-substrate interactions through modeling, to investigate the incorporation of substrates with different degrees of saturation of the malonic acid side chain. This allowed the engineered biosynthesis of new erythromycin derivatives and the introduction of additional mutations into the AT domain for a further shift of the enzyme's substrate scope. Our approach yields non-native polyketide structures with functional groups that will simplify future derivatization approaches, and provides a blueprint for the engineering of AT domains to achieve efficient polyketide synthase diversification.
authors.affiliation: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Kenny
authors.initials: K
authors.lastname: Bravo-Rodriguez
authors.affiliation: Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Klopries
authors.affiliation: Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany.
authors.forename: Kyra R M
authors.initials: KRM
authors.lastname: Koopmans
authors.affiliation: Dr. Fooke-Achterrath Laboratorien GmbH, Habichtweg 16, 41468 Neuss, Germany.
authors.forename: Uschi
authors.initials: U
authors.lastname: Sundermann
authors.affiliation: Université de Caen Basse-Normandie, Centre d'Etudes et de Recherche sur le Médicament de Normandie, 14032 Caen, France.
authors.forename: Samir
authors.initials: S
authors.lastname: Yahiaoui
authors.affiliation: Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Arens
authors.affiliation: Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany.
authors.forename: Susanna
authors.initials: S
authors.lastname: Kushnir
authors.affiliation: Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany. Electronic address: frank.schulz@rub.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Schulz
authors.affiliation: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany. Electronic address: esanchez@mpi-muelheim.mpg.de.
authors.forename: Elsa
authors.initials: E
authors.lastname: Sanchez-Garcia
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism; D004917:Erythromycin / Q000031:analogs & derivatives / Q000096:biosynthesis; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D048630:Polyketide Synthases / Q000737:chemistry; D061065:Polyketides / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
chemical_list: D000900:Anti-Bacterial Agents; D061065:Polyketides; D004917:Erythromycin; D048630:Polyketide Synthases; D000217:Acyltransferases