title: Genomic mutational analysis of the impact of the classical strain improvement program on β-lactam producing Penicillium chrysogenum.
abstract: BACKGROUND
  Penicillium chrysogenum is a filamentous fungus that is employed as an industrial producer of β-lactams. The high β-lactam titers of current strains is the result of a classical strain improvement program (CSI) starting with a wild-type like strain more than six decades ago. This involved extensive mutagenesis and strain selection for improved β-lactam titers and growth characteristics. However, the impact of the CSI on the secondary metabolism in general remains unknown.
  
  
  RESULTS
  To examine the impact of CSI on secondary metabolism, a comparative genomic analysis of β-lactam producing strains was carried out by genome sequencing of three P. chrysogenum strains that are part of a lineage of the CSI, i.e., strains NRRL1951, Wisconsin 54-1255, DS17690, and the derived penicillin biosynthesis cluster free strain DS68530. CSI has resulted in a wide spread of mutations, that statistically did not result in an over- or underrepresentation of specific gene classes. However, in this set of mutations, 8 out of 31 secondary metabolite genes (20 polyketide synthases and 11 non-ribosomal peptide synthetases) were targeted with a corresponding and progressive loss in the production of a range of secondary metabolites unrelated to β-lactam production. Additionally, key Velvet complex proteins (LeaA and VelA) involved in global regulation of secondary metabolism have been repeatedly targeted for mutagenesis during CSI. Using comparative metabolic profiling, the polyketide synthetase gene cluster was identified that is responsible for sorbicillinoid biosynthesis, a group of yellow-colored metabolites that are abundantly produced by early production strains of P. chrysogenum.
  
  
  CONCLUSIONS
  The classical industrial strain improvement of P. chrysogenum has had a broad mutagenic impact on metabolism and has resulted in silencing of specific secondary metabolite genes with the concomitant diversion of metabolism towards the production of β-lactams.
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands. o.v.salo@rug.nl.
authors.forename: Oleksandr V
authors.initials: OV
authors.lastname: Salo
authors.affiliation: Division of Analytical Biosciences, Leiden/Amsterdam Center for Drug Research, Leiden, The Netherlands. marco.ries@gmx.de.|Netherlands Metabolomics Centre, Leiden University, Leiden, The Netherlands. marco.ries@gmx.de.
authors.forename: Marco
authors.initials: M
authors.lastname: Ries
authors.affiliation: Bioinformatics, Wageningen University, Wageningen, The Netherlands. marnix.medema@wur.nl.
authors.forename: Marnix H
authors.initials: MH
authors.lastname: Medema
authors.affiliation: DSM Biotechnology Centre, Delft, The Netherlands. Peter.Lankhorst@dsm.com.
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lankhorst
authors.affiliation: Division of Analytical Biosciences, Leiden/Amsterdam Center for Drug Research, Leiden, The Netherlands. robvreeken@gmail.com.|Netherlands Metabolomics Centre, Leiden University, Leiden, The Netherlands. robvreeken@gmail.com.
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Vreeken
authors.affiliation: DSM Biotechnology Centre, Delft, The Netherlands. Roel.Bovenberg@DSM.COM.|Synthetic Biology and Cell Engineering, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands. Roel.Bovenberg@DSM.COM.
authors.forename: Roel A L
authors.initials: RA
authors.lastname: Bovenberg
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands. a.j.m.driessen@rug.nl.|Kluyver Centre for Genomics of Industrial Fermentations, Julianalaan 67, 2628BC, Delft, The Netherlands. a.j.m.driessen@rug.nl.
authors.forename: Arnold J M
authors.initials: AJ
authors.lastname: Driessen
mesh_terms: D005800:Genes, Fungal; D055442:Metabolome; D005810:Multigene Family; D009154:Mutation; D010408:Penicillium chrysogenum / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D047090:beta-Lactams / Q000378:metabolism*
chemical_list: D047090:beta-Lactams