Estudo químico de metabólitos especiais produzidos por fungos endofíticos de Moringa oleifera Lam
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Data
2019-09-27
Autores
Braun, Poliana Cardoso
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Universidade Federal de Mato Grosso
Resumo
Increased resistance of microorganisms to commonly used antibiotics has been a major concern
in both human health and agriculture. In Brazil, over-resistant microorganisms (sepsis) in
hospitals are the leading cause of death from generalized hospital infections, being considered
a public health problem. In agriculture the fight against phytopathogenic microorganisms has
been causing several environmental impacts due to the improper and exaggerated use of
agrochemicals. Therefore, the present work aims to isolate and elucidate molecules with
antibiotic potential from endophytic fungi associated with the Moringa oleifera Lam. species.32
fungi strains were isolated from different parts of the plant (leaf, stem, seed and flower).
Endophytes were grown for seven days on a small scale with 05 Petri dishes in PDA (PotatoDextrose-Agar) medium, randomly cut and transferred with culture medium to 500 mL
Erlenmeyers flask with 100 mL ethyl acetate. After 24 hours of extraction on the orbital shaker
at 190 rpm, and one hour of sonication, the organic phase was subjected to vacuum evaporator
rotation, obtaining the crude extracts. These were solubilized with methanol:water (95:5) at a
concentration of 20 mg/mL, subjected to solid phase extraction in Sep-Pak cartridge (1g silica
C-18) and analyzed by Ultra Fast Liquid Chromatography - Photodiode Detector Array (UFLCPDA) in exploratory gradient method [analytical C18 column, methanol:water (5:95) to
(100:0), 40']. Subsequently, 21 strains were tested and subjected to antimicrobial agar diffusion
assays against five strains of bacteria (Staphylococcus aureus ATCC 25923, Escherichia coli
ATCC 25922, Klebsiella pneumoniae ATCC 13883, Pseudomonas aeruginosa ATCC 27853
and Enterococcus faecalis ATCC 29292), and five yeast fungi (Candida albicans ATCC 10231,
Candida krusei ATCC 6258, Candida tropicalis ATCC 25922, Candida glabrata ATCC 90030
and Candida parapsilosis ATCC 40058), human pathogens, respectively, cultured at 37ºC for
24h (bacteria) and 48h (fungi), and thereafter, aqueous suspensions were prepared from the
grown colonies following the MacFarland 0.5 (fungus) and 1.0 (bacteria) standards. The
preliminary chromatogram analysis allowed the selection of the fungi MOS02, MOF32,
MOC07 and MOC09 for continuity of the work. These were again cultivated on a larger scale
in PDA (50 Petri dishes), yielding 1493.5 mg (crude extract); 638.0 mg; 1040.5 mg; and 604.0
mg, respectively, after extraction with EtOAc and drying. There were no promising results
regarding antibiosis tests against human pathogens.
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BRAUN, Poliana Cardoso. Estudo químico de metabólitos especiais produzidos por fungos endofíticos de Moringa oleifera Lam. 2019. 50 f. Dissertação (Mestrado em Gestão e Tecnologia Ambiental) - Universidade Federal de Mato Grosso, Instituto de Ciências Agrárias e Tecnológicas, Rondonópolis, 2019.