Caracterização de compostos antimicrobianos produzidos por Streptomyces sp

Abstract

The increase of antibiotic-resistant bacteria encourages the search for new antibacterial substances. Therefore, the selection of microorganisms with potential for the production of new antimicrobial compounds has been extensively studied. Among these microorganisms, special attention is given to actinomycetes that are capable of producing a wide variety of bioactive compounds, such as antibiotics, antifungals, antitumorals and other compounds that can be applied in various segments of the industry. The genus Streptomyces is considered of great industrial importance due to its capacity to produce many secondary metabolites, accounting for 80% of the antibiotics currently used. In previous studies, it was possible to report the antimicrobial activity of a strain of Streptomyces spp. Which showed inhibitory activity against Staphylococcus aureus ATCC 25923, Streptococcus pneumoniae ATCC 49619 and Enterococcus faecalis ATCC 292123. Subsequently, its tuberculicidal activity was also inhibited by the growth of M. tuberculosis H37Rv (ATCC 27294). From these preliminary results, the present work had the objective of analyzing extracts and fractions of metabolites of Streptomyces sp. Aiming to isolate, purify and chemically identify the compound (s) with antimicrobial and tuberculicidal activity. The fractions in hexane (fr-Hex), ethyl acetate (fr-AcoEt) and chloroform (fr-Clo) were obtained from the metabolites obtained in liquid AC medium and by liquid-liquid partition using solvents of increasing polarity. The fr-Clo was chosen to continue the tests because its yield was higher, presenting high antibacterial activity in the Minimum Inhibitory Concentration (MIC) tests against the bacteria Staphylococcus aureus and Mycobacterium smegmatis. Thus, the fr-Clo was subjected to thin layer chromatography (CCD) using several eluent systems and then the Bioautography technique was performed to identify the compounds with activity. From this purification two bioactive sub-fractions were obtained, which were subjected to the isolation of the compounds by preparative CCD and HPLC-DAD-MS technique. The isolated substances were named Clarin A and Clarin B, which presented a Minimum Inhibitory Concentration (MIC) of 1.0 μg / mL for Clarin A and 0.5 μg / mL for Clarin B against Staphylococcus aureus and 16 μg / Ml against Mycobacterium smegmatis. They did not present cytotoxicity to 3T3-L1 non-tumor cells. The Clarin A and Clarin B substances were analyzed using Nuclear Magnetic Resonance (NMR) and low resolution and high resolution Mass Spectrometry techniques for chemical identification. The isolated substances were identified as Actinomycin D and Actinomycin X2 with m / z of 1277 [M + Na] and 1291 [M + Na]. The strong antibacterial activity of the isolated substances makes this product an important source of natural antibacterial compounds.