Prospecção de metabólitos de Aspergillus spp. e Penicillium spp. da coleção de fungos da Amazônia (CFAM) para atividade antimicrobiana e produção de proteases

Abstract

The exploration of new natural antibiotics is a time-consuming and high challenge. costs. The pharmaceutical industry has largely abandoned these compounds in favor of the large number of synthetic molecules already known and described in the literature. However, natural antibiotics produced by microorganisms have some features that make the search for these molecules necessary. They are original molecules, with an immense diversity of chemical structures and biological activities. it is known that fungi isolated from different substrates of the Amazon biome are microorganisms producers of biomolecules with therapeutic potential and, among such fungi, we can highlight the genera Aspergillus and Penicillium. Such genres are also known as produce enzymes of biotechnological interest, such as proteases, which are used in different segments of the industry. Thus, the study aimed to evaluate the antimicrobial and proteolytic potential of filamentous fungi stored in the Collection of Amazon Fungi - CFAM, as well as characterize the chemical profile of fungi with best activities. One hundred fungal cultures were selected, being 50 of Aspergillus spp. and 50 of Penicillium spp., stored in the Amazon Fungi Collection – CFAM of the ILMD/FIOCRUZ. The reactivation of cultures was carried out according to the literature specialized. To obtain the extracts, viable cultures were cultivated in Agar Yeast and Sucrose Extract (YES), at 28°C and, after seven days, cold extraction was performed of biocompounds in Ethyl Acetate. The extracted biocompounds were tested in front Candida albicans CFAM 1286, Staphylococcus aureus CBAM 0324 and Escherichia coli CBAM 0002, by the "cup plate" method and the Minimum and Inhibitory Concentration was performed (CIM). The extracts that showed activity were subjected to CIM and those that presented the lowest concentrations in this (0.156 and 0.312 mg/mL) followed by the characterization of the chemical profile by Thin Layer Chromatography (TLC) and respective physical (with UV light of 254 and 365 nm), chemical (using Dragendorf, Anisaldehyde, Ferric Chloride, Ceric Sulfate and NP/PEG) and biological (bioautography with E. coli 0002, S. aureus 0324 and C. albicans 1286) and they were also evaluated for their proteolytic production where the fungal fragments were cultivated in manachini solution for 5 days at 28°C, where the tests on "Cup plate" and reading of enzyme production through the spectrophotometer. Among the extracts obtained from the 100 selected fungal cultures, 64% (n=64) of the extracts showed positive activity for at least one microorganism tested with inhibition halos ranging from 2 to 30 mm in diameter. At CIM, of the 64 extracts that showed activity, 24% (n=15) showed activity at the lowest concentrations of the study (0.156 and 0.312 mg/mL) and these went to the CCD, where it was possible to notice a wide variety of compounds found. Of the 15 fungi, five were lost by contamination, with only ten being used for enzyme production. In the tests of "Cup plate", only one fungal filtrate did not show activity. The studied fungi showed satisfactory activity both for antimicrobials and for the production of proteases, contributing to the better characterization of the fungal species in the region Amazon, specifically the genera Aspergillus and Penicillium, as well as for the advance in the biotechnology sector. The fungal extracts studied showed results promising, however future tests are needed to better elucidate the compounds. KEYWORDS: Antimicrobial; enzymes; chemical compounds.