title: A branched biosynthetic pathway is involved in production of roquefortine and related compounds in Penicillium chrysogenum.
abstract: Profiling and structural elucidation of secondary metabolites produced by the filamentous fungus Penicillium chrysogenum and derived deletion strains were used to identify the various metabolites and enzymatic steps belonging to the roquefortine/meleagrin pathway. Major abundant metabolites of this pathway were identified as histidyltryptophanyldiketopiperazine (HTD), dehydrohistidyltryptophanyldi-ketopiperazine (DHTD), roquefortine D, roquefortine C, glandicoline A, glandicoline B and meleagrin. Specific genes could be assigned to each enzymatic reaction step. The nonribosomal peptide synthetase RoqA accepts L-histidine and L-tryptophan as substrates leading to the production of the diketopiperazine HTD. DHTD, previously suggested to be a degradation product of roquefortine C, was found to be derived from HTD involving the cytochrome P450 oxidoreductase RoqR. The dimethylallyltryptophan synthetase RoqD prenylates both HTD and DHTD yielding directly the products roquefortine D and roquefortine C without the synthesis of a previously suggested intermediate and the involvement of RoqM. This leads to a branch in the otherwise linear pathway. Roquefortine C is subsequently converted into glandicoline B with glandicoline A as intermediates, involving two monooxygenases (RoqM and RoqO) which were mixed up in an earlier attempt to elucidate the biosynthetic pathway. Eventually, meleagrin is produced from glandicoline B involving a methyltransferase (RoqN). It is concluded that roquefortine C and meleagrin are derived from a branched biosynthetic pathway.
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Zernike Institute for Advanced Materials, University of Groningen, Groningen, The Netherlands.
authors.forename: Hazrat
authors.initials: H
authors.lastname: Ali
authors.forename: Marco I
authors.initials: MI
authors.lastname: Ries
authors.forename: Jeroen G
authors.initials: JG
authors.lastname: Nijland
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lankhorst
authors.forename: Thomas
authors.initials: T
authors.lastname: Hankemeier
authors.forename: Roel A L
authors.initials: RA
authors.lastname: Bovenberg
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Vreeken
authors.forename: Arnold J M
authors.initials: AJ
authors.lastname: Driessen
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics* / Q000502:physiology*; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers / Q000235:genetics; D006576:Heterocyclic Compounds, 4 or More Rings / Q000378:metabolism; D007211:Indoles / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D046228:Microarray Analysis; D005810:Multigene Family / Q000235:genetics*; D010059:Ovomucin / Q000096:biosynthesis*; D010408:Penicillium chrysogenum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D010879:Piperazines / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D006576:Heterocyclic Compounds, 4 or More Rings; D007211:Indoles; D010879:Piperazines; D010059:Ovomucin; C060672:meleagrin; C012536:roquefortine