Estratégias de produção metabólica e aplicação biotecnológica de fungos endofíticos isolados de Clitoria guianensis

Abstract

The co-culture strategy aims to mimic environmental conditions of interactions between microorganisms, thus improving the production of secondary metabolites of interest. This study intends to use the co-culture technique between the endophytic fungi Neopestalotiopsis sp. Cg13, Fusarium sp. Cg43 and Fusarium sp. Cg45, isolated from Clitoria guianensis, a common plant in the Cerrado of Tocantins, and between the endophytes and the phytopathogen Sclerotinia sclerotiorum, aiming to analyze how microbial interaction affects the production of antimicrobial and cytotoxic compounds. Using the GNPS2 platform (Global Natural Product Social Molecular Networking 2), a molecular network was created with the mass spectrometry data obtained from the monocultures and co-cultures, enabling the visualization, analysis and annotation of compounds, the fractions related to microbial interactions between Neopestalotiopsis sp. Cg13 and Fusarium sp. Cg45, and Neopestalotiopsis sp. Cg13 and S. sclerotiorum showed greater chemical diversity when compared to the other cocultures, however, the monocultures S. sclerotiorum and Fusarium sp. Cg45 had the richest profile among all fractions. A total of 35 compounds were recorded at different levels, in which the class of most compounds could be attributed to lipids, in addition, the chemical structure of 14 metabolites was determined. The study demonstrated that the co-cultures stimulated the production of secondary metabolites with antimicrobial activity, and the 45xSS fraction showed antifungal activity against S. sclerotiorum, suggesting that the endophyte-pathogen interaction is essential for the production of antagonistic substances. In the antibacterial tests, only the endophyte-endophyte cocultures showed the ability to inhibit the growth of the pathogenic bacteria Stenotrophomonas maltophilia and Acinetobacter baumannii, with fractions 43x45, 13x45 and 13x43. Furthermore, none of the fractions showed significant cytotoxic activity against human HaCaT and HCT-116 cells, indicating the absence of relevant toxic compounds. Since there is no cytotoxicity, the use of microorganisms and fractions does not pose risks to human health if used in the agricultural and pharmaceutical sectors.