Evidencing 98 secondary metabolites of Penicillium verrucosum using substrate isotopic labeling and high-resolution mass spectrometry

Thaïs Hautbergue (1) , Olivier Puel (2) , Souria Tadrist (1) , Lauriane Meneghetti , Michel Péan (3, 4) , Marcel Delaforge (5) , Laurent Debrauwer (6) , Isabelle P. Oswald (2) , Emilien L. Jamin (6)

1 ToxAlim - ToxAlim
2 ToxAlim-BioToMyc - Biosynthèse & Toxicité des Mycotoxines
3 GRAP - Groupe de Recherches Appliquées en Phytotechnologie
4 MEM - Microbiologie Environnementale et Moléculaire
5 SMPSD - Système membranaires, photobiologie, stress et détoxication
6 E20 - MetaToul AXIOM

Industrial applications of fungal compounds, coupled with the emergence of fungal threats to naturalecosystems and public health, have increased interest in filamentous fungi. Among all pathogenic fungi,Penicillium verrucosum is one of the most common mold-infecting stored cereals in temperate regions.However, it is estimated that 80% of fungal secondary metabolites remain unknown. To detect new P.verrucosum compounds, an untargeted metabolomic approach was applied to fungus grown on wheatgrains labeled with stable isotopes: (i) natural grains (99%12C); (ii) grains enriched with 97% of13C; and(iii) grains enriched with 53% of13C and 97% of15N. Analyses performed by high-performance liquidchromatography coupled with high-resolution mass spectrometry (HPLC–HRMS) enabled the specificdetection of fungal metabolites, and the unambiguous characterization of their chemical formulas. Inthis way, 98 secondary metabolites were detected and their chemical formulas were determined. Ofthese, only 18 identifications could be made based on databases, the literature and mass spectrometryfragmentation experiments, with the result that 80 were totally unknown. Molecular networks weregenerated to analyze these results, leading to the characterization by MSnexperiments of a new fungis-porin produced by P. verrucosum. More generally, this article provides precise mass spectrometric dataabout all these compounds for further studies of the Penicillium metabolome.

Mots clés: secondary plant products; mass spectrum analysis; secondary metabolite; molecular networking; public health; fungus; mass spectrometry; stable isotope labeling

Mots clés: toxicologie alimentaire; penicillium verrucosum; santé publique; spectrométrie de masse; métabolite secondaire

Domaines: Sciences du Vivant [q-bio]

Thaïs Hautbergue, Olivier Puel, Souria Tadrist, Lauriane Meneghetti, Michel Péan, et al.. Evidencing 98 secondary metabolites of Penicillium verrucosum using substrate isotopic labeling and high-resolution mass spectrometry. Journal of Chromatography B - Analytical Technologies in the Biomedical and Life Sciences, 2017, in press, ⟨10.1016/j.jchromb.2017.03.011⟩. ⟨hal-01607721⟩