Jørgensen, Simon Hartung5; Frandsen, Rasmus John Normand6; Nielsen, Kristian Fog7; Lysøe, Erik8; Sondergaard, Teis Esben5; Wimmer, Reinhard5; Giese, Henriette5; Sørensen, Jens Laurids5
1 Department of Systems Biology, Technical University of Denmark2 Metabolomics Platform, Department of Systems Biology, Technical University of Denmark3 Metabolic Signaling and Regulation, Department of Systems Biology, Technical University of Denmark4 Eucaryotic Molecular Cell Biology, Department of Systems Biology, Technical University of Denmark5 Aalborg University6 Biosynthetic Pathway Engineering, Department of Biotechnology and Biomedicine, Technical University of Denmark7 DTU Metabolomics Core, Department of Biotechnology and Biomedicine, Technical University of Denmark8 Norwegian Institute for Agricultural and Environmental Research
The available genome sequences show that the number of secondary metabolite genes in filamentous fungi vastly exceeds the number of known products. This is also true for the global plant pathogenic fungus Fusarium graminearum, which contains 15 polyketide synthase (PKS) genes, of which only 6 have been linked to products. To help remedy this, we focused on PKS14, which has only been shown to be expressed during plant infections or when cultivated on rice or corn meal (RM) based media. To enhance the production of the resulting product we introduced a constitutive promoter in front of PKS14 and cultivated two of the resulting mutants on RM medium. This led to the production of two compounds, which were only detected in the PKS14 overexpressing mutants and not in the wild type or PKS14 deletion mutants. The two compounds were tentatively identified as orsellinic acid and orcinol by comparing spectroscopic data (mass spectroscopy and chromatography) to authentic standards. NMR analysis of putative orcinol isolated from the PKS14 overexpressing mutant supported our identification. Orcinol and orsellinic acid, not previously detected in Fusarium, have primarily been detected in lichen fungi. Orsellinic acid is hypothesized to be the PKS release product which is transformed to orcinol through decarboxylation. Phylogenetic analyses of PKSs placed PKS14 in a subclade of known OA synthases. Expression analysis by microarray of 55 experiments identified seven genes near PKS14 that were expressed in a similar manner. One of the seven genes encodes a predicted carboxylase, which could be responsible for transforming orsellinic acid to orcinol.
Fungal Genetics and Biology, 2014, Vol 70, p. 24-31