title: Genetic engineering activates biosynthesis of aromatic fumaric acid amides in the human pathogen Aspergillus fumigatus.
abstract: The Aspergillus fumigatus nonribosomal peptide synthetase FtpA is among the few of this species whose natural product has remained unknown. Both FtpA adenylation domains were characterized in vitro. Fumaric acid was identified as preferred substrate of the first and both l-tyrosine and l-phenylalanine as preferred substrates of the second adenylation domain. Genetically engineered A. fumigatus strains expressed either ftpA or the regulator gene ftpR, encoded in the same cluster of genes, under the control of the doxycycline-inducible tetracycline-induced transcriptional activation (tet-on) cassette. These strains produced fumaryl-l-tyrosine and fumaryl-l-phenylalanine which were identified by liquid chromatography and high-resolution mass spectrometry. Modeling of the first adenylation domain in silico provided insight into the structural requirements to bind fumaric acid as peptide synthetase substrate. This work adds aromatic fumaric acid amides to the secondary metabolome of the important human pathogen A. fumigatus which was previously not known as a producer of these compounds.
authors.affiliation: Department of Pharmaceutical Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Friedrich-Schiller-Universität, Jena, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kalb
authors.affiliation: Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Jena, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Heinekamp
authors.affiliation: Department of Pharmaceutical Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Friedrich-Schiller-Universität, Jena, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Lackner
authors.affiliation: Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Jena, Germany.
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Scharf
authors.affiliation: Department of Infection Biology, Leibniz Institute for Natural Product Research and Infection Biology, Jena, Germany.
authors.forename: Hans-Martin
authors.initials: HM
authors.lastname: Dahse
authors.affiliation: Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Jena, Germany.
authors.forename: Axel A
authors.initials: AA
authors.lastname: Brakhage
authors.affiliation: Department of Pharmaceutical Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Friedrich-Schiller-Universität, Jena, Germany dirk.hoffmeister@hki-jena.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Hoffmeister
mesh_terms: D000577:Amides / Q000378:metabolism*; D001232:Aspergillus fumigatus / Q000235:genetics* / Q000378:metabolism*; D002853:Chromatography, Liquid; D005650:Fumarates / Q000378:metabolism*; D015870:Gene Expression; D013058:Mass Spectrometry; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D005810:Multigene Family; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D000577:Amides; D005650:Fumarates; D011994:Recombinant Proteins; C032005:fumaric acid