Abstract
There is a growing imperative for research into alternative compounds for the treatment of the fungal infections. Thus, many studies have focused on the analysis of antifungal proteins and peptides from different plant sources. Among these molecules are protease inhibitors (PIs). Previously, PIs present in the peptide-rich fractions called PEF1, PEF2 and PEF3 were identified from Capsicum chinense seeds, which have strong activity against phytopathogenic fungi. The aim of this study was to evaluate the mechanism of action and antimicrobial activity of PIs from PEF2 and PEF3 on the growth of yeasts of the genus Candida. In this work, analyses of their antimicrobial activity and cell viability were carried out. Subsequently, the mechanism of action by which the PIs cause the death of the yeasts was evaluated. Cytotoxicity was assessed in vitro by erythrocytes lysis and in vivo in Galleria mellonella larvae. PEF2 and PEF3 caused 100% of the growth inhibition of C. tropicalis and C. buinensis. For C. albicans inhibition was approximately 60% for both fractions. The PEF2 and PEF3 caused a reduction in mitochondrial functionality of 54% and 46% for C. albicans, 26% and 30% for C. tropicalis, and 71% and 68% for C. buinensis, respectively. These fractions induced morphological alterations, led to membrane permeabilization, elevated ROS levels, and resulted in necrotic cell death in C. tropicalis, whilst demonstrating low toxicity toward host cells. From the results obtained here, we intend to contribute to the understanding of the action of PIs in the control of fungal diseases of medical importance.
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Acknowledgements
This study forms a part of the postdoctoral MSS and was carried out at the Universidade Estadual do Norte Fluminense. We wish to thank L.C.D. Souza and V.M. Kokis for technical assistance.
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This study was financed in part by the Universidade Estadual do Norte Fluminense, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil(CAPES) – Finance Code 001. We acknowledge the financial support from the Brazilian agencies CNPq (307590/2021-6; 400724/2023-5) FAPERJ (E-26/200567/2023; E-26/210.353/2022; E-26/210.625/2023).
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All the authors contributed extensively to the work presented in this manuscript. The study was conceived by MSS and VMG. Experimental procedures were carried out by MSS, GBT, RSG, LAS, MBC, TAMS, FFM and PSR. Data analyses were performed by MSS, GBT, OVM, SHS, RR, and VMG. The paper was written by MSS, GBT, AOC and VMG.
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Silva, M.S.d., Taveira, G.B., Silva Gebara, R.d. et al. Understanding the mechanism of action of protease inhibitors in controlling the growth of the Candida Genus: potential candidates for development of new antifungal molecules. Arch Microbiol 206, 257 (2024). https://doi.org/10.1007/s00203-024-03993-7
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DOI: https://doi.org/10.1007/s00203-024-03993-7