Biochemical characterization of a phospholipase A2 homologue from the venom of the social wasp Polybia occidentalis

Background: Wasp venoms constitute a molecular reservoir of new pharmacological substances such as peptides and proteins, biological property holders, many of which are yet to be identified. Exploring these sources may lead to the discovery of molecules hitherto unknown. This study describes, for the first time in hymenopteran venoms, the identification of an enzymatically inactive phospholipase A2 (PLA2) from the venom of the social wasp Polybia occidentalis. Methods: P. occidentalis venom was fractioned by molecular exclusion and reverse phase chromatography. For the biochemical characterization of the protein, 1D and 2D SDS-PAGE were performed, along with phospholipase activity assays on synthetic substrates, MALDI-TOF mass spectrometry and sequencing by Edman degradation. Results: The protein, called PocTX, was isolated using two chromatographic steps. Based on the phospholipase activity assay, electrophoresis and mass spectrometry, the protein presented a high degree of purity, with a mass of 13,896. 47 Da and a basic pI. After sequencing by the Edman degradation method, it was found that the protein showed a high identity with snake venom PLA2 homologues. Conclusion: This is the first report of an enzymatically inactive PLA2 isolated from wasp venom, similar to snake PLA2 homologues.

Biochemical characterization of a phospholipase A2 homologue from the venom of the social wasp Polybia occidentalis

Wasp venoms constitute a molecular reservoir of new pharmacological substances such as peptides and proteins, biological property holders, many of which are yet to be identified. Exploring these sources may lead to the discovery of molecules hitherto unknown. This study describes, for the first time in hymenopteran venoms, the identification of an enzymatically inactive phospholipase A2 (PLA2) from the venom of the social wasp Polybia occidentalis. Methods: P. occidentalis venom was fractioned by molecular exclusion and reverse phase chromatography. For the biochemical characterization of the protein, 1D and 2D SDS-PAGE were performed, along with phospholipase activity assays on synthetic substrates, MALDI-TOF mass spectrometry and sequencing by Edman degradation. Results: The protein, called PocTX, was isolated using two chromatographic steps. Based on the phospholipase activity assay, electrophoresis and mass spectrometry, the protein presented a high degree of purity, with a mass of 13,896. 47 Da and a basic pI. After sequencing by the Edman degradation method, it was found that the protein showed a high identity with snake venom PLA2 homologues. Conclusion: This is the first report of an enzymatically inactive PLA2 isolated from wasp venom, similar to snake PLA2 homologues.(AU)

Antifungal Action of the Dillapiole-rich Oil of Piper aduncum against Dermatomycoses Caused by Filamentous Fungi.

Aims and Study Design: Piper aduncum L. is a Brazilian plant with many biological properties attributed to its dillapiole-rich essential oil. Despite the development of antibiotics, bacterial and fungal infections are still a public health issue in the medical field. This study measured the antimicrobial activity of the dillapiole-rich essential oil of P. aduncum against pathogenic skin microorganisms. Place of Study: Faculty of Pharmacy and Graduate Program in Pharmaceutical Science, Federal University of Pará, Brazil. This work was performed in 2014. Methodology: Gas Chromatography (GC) and Gas Cromatography-Mass spectrometry (GC-MS) have analyzed the oil and its dillapiole-rich fraction. The determination of Minimum Inhibitory Concentration (MIC) and Minimum fungicidal concentration (MFC) values was carried out by microdilution method and counting of formed colonies. Results: For the strains of Trichophyton mentagrophytes (ATCC 9533 and clinical isolate), the oil and its dillapiole-rich fraction exhibited MIC values of 500 μg/ml while the MFC values were 1,500 μg/ml for the oil and 1,000 μg/ml for the fraction rich in dillapiole. For clinical isolates of T. rubrum and Epidermophyton floccosum, MIC values of 500 μg/ml and MFC 1,500 μg/ml were equal for the oil and the dillapiole-rich fraction, respectively. For clinical isolates of Microsporum canis and M. gypseum, the MIC and MFC values were 250 μg/ml and 500 μg/ml, respectively. For strains of Aspergillus fumigatus (ATCC 40152 and clinical isolate), the oil and its dillapiole-rich fraction have shown the same MIC value of 3.9 μg/ml while the MFC values were 7.8 μg/ml for the strain ATCC 40152, and 15.6 μg/ml for the clinical isolate. The oil and dillapiole-rich fraction did not show antibacterial activity against the strain of Staphylococcus aureus ATCC 6538 and its clinical isolate Conclusion: The dillapiole-rich essential oil of P. aduncum and its dillapiole-rich fraction demonstrates significant antifungal activity against dermatophytes, filamentous fungi and potent antifungal activity against non-dermatophyte filamentous fungi.

Antibacterial activity of the Piper aduncum oil and dillapiole, its main constituent, against multidrug-resistant strains / Actividad antibacteriana del aceite de Piper aduncum y dillapiole, su componente principal, frente a cepas resistentes a múltiples fármacos

The study aimed to evaluate the bactericidal activity of oil essential and dillapiole from P. aduncum against standard and multidrug-resistant strains of Staphylococcus spp. The oil showed antimicrobial action against these strains, but better results were obtained for the standards strains of S. epidermidis and S. aureus, with MIC of 250 and 500 ug/mL, respectively. Dillapiolle was less effective than the oil against the same standard and multi-drug resistant strains (MIC =1000 ug/mL). However, when dillapiolle was tested in combination with myristicin, another component of the oil, it increased its bactericidal activity and showed a synergistic action...

El objetivo del estudio fue evaluar la actividad bactericida de los aceites esenciales y dillapiole de P. aduncum contra cepas estándar y multirresistentes de Staphylococcus spp. El aceite mostró acción antimicrobiana frente a estas cepas, pero se obtuvo mejores resultados para las cepas de S. epidermidis y S. aureus, con MIC de 250 y 500 ug/ml, respectivamente. Dillapiolle fue menos eficaz que el aceite contra cepas estándar y multirresistentes (MIC = 1000 ug/ml). Sin embargo, cuando dillapiolle fue probado en combinación con la miristicina, otro componente del aceite, que aumentó su actividad bactericida y mostró una acción sinérgica...