title: Solving the Puzzle of One-Carbon Loss in Ripostatin Biosynthesis.
abstract: Ripostatin is a promising antibiotic that inhibits RNA polymerase by binding to a novel binding site. In this study, the characterization of the biosynthetic gene cluster of ripostatin, which is a peculiar polyketide synthase (PKS) hybrid cluster encoding cis- and trans-acyltransferase PKS genes, is reported. Moreover, an unprecedented mechanism for phenyl acetic acid formation and loading as a starter unit was discovered. This phenyl-C2 unit is derived from phenylpyruvate (phenyl-C3) and the mechanism described herein explains the mysterious loss of one carbon atom in ripostatin biosynthesis from the phenyl-C3 precursor. Through in vitro reconstitution of the whole loading process, a pyruvate dehydrogenase like protein complex was revealed that performs thiamine pyrophosphate dependent decarboxylation of phenylpyruvate to form a phenylacetyl-S-acyl carrier protein species, which is supplied to the subsequent biosynthetic assembly line for chain extension to finally yield ripostatin.
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.
authors.forename: Chengzhang
authors.initials: C
authors.lastname: Fu
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Auerbach
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.|Current address: Laboratory Molecules of Communication and Adaptation of Microorganisms (MCAM, UMR 7245 CNRS-MNHN), Sorbonne Universités, Muséum National d'Histoire Naturelle, Centre National de la Recherche Scientifique, CP 54, 57 rue Cuvier, 75005, Paris, France.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.|German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Braunschweig, Germany.
authors.forename: Ullrich
authors.initials: U
authors.lastname: Scheid
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.|German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Braunschweig, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Luxenburger
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.|German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Braunschweig, Germany.
authors.forename: Ronald
authors.initials: R
authors.lastname: Garcia
authors.affiliation: Helmholtz Centre for Infection Research (HZI), Inhoffenstrasse 7, 38124, Braunschweig, Germany.
authors.forename: Herbert
authors.initials: H
authors.lastname: Irschik
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, Campus Building E8.1, 66123, Saarbrücken, Germany.|German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Braunschweig, Germany.
authors.forename: Rolf
authors.identifier: 0000-0001-6571-6630
authors.initials: R
authors.lastname: Müller
mesh_terms: D000213:Acyl Carrier Protein / Q000235:genetics / Q000378:metabolism; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D053898:Biosynthetic Pathways; D005798:Genes, Bacterial; D007783:Lactones / Q000378:metabolism*; D005810:Multigene Family; D009231:Myxococcales / Q000235:genetics / Q000378:metabolism*; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism
chemical_list: D000213:Acyl Carrier Protein; D000900:Anti-Bacterial Agents; D007783:Lactones; C096600:ripostatin A; D048630:Polyketide Synthases; D000217:Acyltransferases
keywords: biosynthesis; phenylpyruvate; polyketide synthases; pyruvate dehydrogenase complex; ripostatin