ALMEIDA, C. A.

ALVIANO, C. S.

ALVIANO, D. S.

AZEVEDO, M. M. B. de

BERGTER, E. B.

BIZZO, H. R.

FREITAS, F. C.

English

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8th Brazilian Symposium on Essential Oils - International Symposium on Essential Oils Poster 080 November 10 to 13, 2015 - Rio de Janeiro Botanical Garden, Rio de Janeiro, Brazil  ISBN: 978-85-66836-11-0   In vitro evaluation of inhibitory activity of some species of Croton and Piper essential oils in secreted proteases of Pseudallescheria boydii  Mariana M. B. de Azevedo1, Catia A. Almeida1, Francisco C. M. Chaves2, Eliana B. Bergter1, Humberto R. Bizzo3, Celuta S. Alviano1, Daniela S. Alviano1  1 Federal Universityof Rio de Janeiro - Rio de Janeiro, Brazil 2 Embrapa Western Amazon – Amazon, Brazil 3 Embrapa Food Technology – Rio de Janeiro, Brazil marimbarros@gmail.com  Keywords: Croton, Piper, essential oil, Pseudallescheria boydii, secreted proteases.   Pseudallescheria boydii is a fungal organism known as a soil and water natural inhabitant and decaying vegetation, which has become an increasingly recognized pathogen among immunocompromised individuals, including HIV infected patients. It is related to cause madura foot and in severe cases, invasive infection of various organs (central nervous, cardiovascular, respiratory systems). One potential drug target for the pharmaceutical industry is proteases. Proteases regulate the fate, localization, and activity of many proteins, modulate protein-protein interactions, create new bioactive molecules, contribute to the processing of cellular information, and generate, transduce, and amplify molecular signals. The aim of this study was to evaluate the inhibitory activity of some species of Croton and Piper essential oils (EOs) in P. boydii secreted proteases. The EOs of 3 species of Croton plants: C. tricolor (sacatinga), C. pulegioides (velandinho) and C. blanchetianus (marmeleiro); and 4 species of Piper plants: P. marginatum (capeba-cheirosa), P. tuberculatum (pimenta darta), P. hispidum (matico-falso) and Piper sp. were obtained by hydrodistillation using a Clevenger-type apparatus for 4 h. The identification of EO was performed by GC/FID and GC/MS in an Agilent 6890N and an Agilent 5973N systems with HP-5MS fused silica capillary columns (30 m X 0.25 mm X 0.25 µm). Hydrogen was used as carrier gas for GC/FID and helium for GC/MS, both with a flow rate of 1.0 mL min-1.Oven temperature was raised from 60 to 240 °C at 3 °C min-1. Mass detector was operated in electronic ionization mode at 70 eV. The minimum inhibitory concentration (MIC) was evaluated in triplicate according standard method from the Clinical and Laboratory Standards Institute (CLSi). To evaluate the activity of Croton and Piper EOs in secreted proteases of P. boydii it was performed assays of proteolytic activity inhibition according to Buroker-Kilgore and Wang. Cell-free supernatants of P. boydii grown in RPMI broth, were incubated with bovine serum albumin (0.1 mg mL-1), as proteic substrate, and some pH buffers. The calculated MIC were >1250 µg mL-1.The preliminary results showed that the extracellular peptidases were able tohydrolizate the proteic substrate on pH 6 in RPMI broth. The proteolytic inhibition obtained ranging of 29 % for P. tuberculatum and 100 % for C. pulegioides and C.tricolor (48 µL mL-1) in RPMI supernatant of P. boydii. These results showed promising activity and suggests a possible target that justifies the antifungal activity of Croton and Piper EOs.  1. Cumbo-Nacheli, G., de Sanctis, J., Holden, D. Am. J. Case Rep., 2012, 13, 163-165. 2. Kenyon, C. et al.South. Afr. J. Epidemiol. Infect., 2011, 26, 88-89. 3. Clinical and Laboratory Standards Institute. M38-A2, 2008. Wayne, PA. 4. Buroker-Kilgore, M., Wang, K.K.W. Anal. Biochem., 1993, 208, 387-392.  Acknowledgements: FAPERJ, CAPES, CNPq. 

In vitro evaluation of inhibitory activity of some species of croton and piper essential oils in secreted proteases of Pseudallescheria boydii.

http://www.alice.cnptia.embrapa.br/bitstream/doc/1029334/1/Resumo8SBOE080CD.pdf

In vitro evaluation of inhibitory activity of some species of croton and piper essential oils in secreted proteases of Pseudallescheria boydii.

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