Estudo metabolômico de Penicillium sp. por meio de espectrometria de massas de alta resolução e redes moleculares

Abstract

Metabolomics is a tool with varied applications and of great importance in screening and discovery of new bioactive compounds of natural origin, using advanced analytical techniques and chemoinformatics. By means of metabolomics, compounds with diverse biotechnological applications were identified and isolated from different organisms. Among the main sources of bioactive compounds, plants and microorganisms stand out. Among the latter, fungi present themselves as sources of metabolites with different biological activities with great economic importance. In this sense, this work aimed to use a metabolomics approach through mass spectrometry and molecular networks for a comprehensive characterization of the secondary metabolites of Penicillium sp. MMSRG-058 and mutant strains of biotechnological interest. To obtain the Penicillium sp., fermentation cultures were carried out by the OSMAC method, using four different culture media (BDL, ISP2, CZAPECK and Carne2), under different conditions, such as static and agitated. Both, the liquid fraction and the mycelium were used, using ethyl acetate and methanol as extracting solvents. Extracts were analyzed using HPLC-HRMS/MS and molecular networks. The analysis of both the networks and the manual interpretation of the MS/MS spectra allowed the identification of more than 30 molecules, most of which as part of polyketides class, specifically azaphilones, among them sclerotioramine, isochromophilone I, II, VI and IX, sclerotiorine and ocrephillone. The production of different molecules in different media and conditions showed a significant difference, with the presence of distinct molecules or even specificity in some cases. In the HPLC-HRMS/MS data of the mutant strains of Penicillium sp., all the polyketides produced by the wild strain were absent, with the exception of the meroterpenoid atlantinone A as the major ion. This same molecule was found in small amounts in only three culture media of the wild strain. A significant number of metabolites still lacks of identification, evidencing the metabolic capacity of Penicillium sp., under different conditions. The study using the metabolomics approach involving analytical techniques and molecular networks allowed the metabolic characterization of the fungus explored, as well as the annotation of molecules of biotechnological interest.