title: Identification and manipulation of the pleuromutilin gene cluster from Clitopilus passeckerianus for increased rapid antibiotic production.
abstract: Semi-synthetic derivatives of the tricyclic diterpene antibiotic pleuromutilin from the basidiomycete Clitopilus passeckerianus are important in combatting bacterial infections in human and veterinary medicine. These compounds belong to the only new class of antibiotics for human applications, with novel mode of action and lack of cross-resistance, representing a class with great potential. Basidiomycete fungi, being dikaryotic, are not generally amenable to strain improvement. We report identification of the seven-gene pleuromutilin gene cluster and verify that using various targeted approaches aimed at increasing antibiotic production in C. passeckerianus, no improvement in yield was achieved. The seven-gene pleuromutilin cluster was reconstructed within Aspergillus oryzae giving production of pleuromutilin in an ascomycete, with a significant increase (2106%) in production. This is the first gene cluster from a basidiomycete to be successfully expressed in an ascomycete, and paves the way for the exploitation of a metabolically rich but traditionally overlooked group of fungi.
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Andy M
authors.initials: AM
authors.lastname: Bailey
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Alberti
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Sreedhar
authors.initials: S
authors.lastname: Kilaru
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Collins
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Kate
authors.initials: K
authors.lastname: de Mattos-Shipley
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Hartley
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Patrick
authors.initials: P
authors.lastname: Hayes
authors.affiliation: GSK, Southdownview Way, Worthing, West Sussex, BN14 8QH, UK.
authors.forename: Alison
authors.initials: A
authors.lastname: Griffin
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Colin M
authors.initials: CM
authors.lastname: Lazarus
authors.affiliation: School of Chemistry, University of Bristol, Cantock's Close Bristol, BS8 1TS, UK.
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Cox
authors.affiliation: School of Chemistry, University of Bristol, Cantock's Close Bristol, BS8 1TS, UK.
authors.forename: Christine L
authors.initials: CL
authors.lastname: Willis
authors.affiliation: GSK, 1250 S. Collegeville Road, Collegeville, Pennsylvania, 19426-0989, United States.
authors.forename: Karen
authors.initials: K
authors.lastname: O'Dwyer
authors.affiliation: School of Chemistry, University of Bristol, Cantock's Close Bristol, BS8 1TS, UK.
authors.forename: David W
authors.initials: DW
authors.lastname: Spence
authors.affiliation: School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, UK.
authors.forename: Gary D
authors.initials: GD
authors.lastname: Foster
mesh_terms: D000363:Agaricales / Q000235:genetics* / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism; D053898:Biosynthetic Pathways / Q000235:genetics*; D003001:Cloning, Molecular; D004224:Diterpenes / Q000378:metabolism; D060847:Metabolic Engineering; D005810:Multigene Family; D011083:Polycyclic Compounds; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000097567:Pleuromutilins
chemical_list: D000900:Anti-Bacterial Agents; D004224:Diterpenes; D011083:Polycyclic Compounds; D011994:Recombinant Proteins; D000097567:Pleuromutilins