Potent Anti-Candida Fraction Isolated from Capsicum chinense Fruits Contains an Antimicrobial Peptide That is Similar to Plant Defensin and is Able to Inhibit the Activity of Different α-Amylase Enzymes
- 17 January 2021
- journal article
- research article
- Published by Springer Nature in Probiotics and Antimicrobial Proteins
- Vol. 13 (3) , 862-872
Abstract
Antimicrobial peptides (AMPs) are molecules present in several life forms, possess broad-spectrum of inhibitory activity against pathogenic microorganisms, and are a promising alternative to combat the multidrug resistant pathogens. The aim of this work was to identify and characterize AMPs from Capsicum chinense fruits and to evaluate their inhibitory activities against yeasts of the genus Candida and α-amylases. Initially, after protein extraction from fruits, the extract was submitted to anion exchange chromatography resulting two fractions. Fraction D1 was further fractionated by molecular exclusion chromatography, and three fractions were obtained. These fractions showed low molecular mass peptides, and in fraction F3, only two protein bands of approximately 6.5 kDa were observed. Through mass spectrometry, we identified that the lowest molecular mass protein band of fraction F3 showed similarity with AMPs from plant defensin family. We named this peptide CcDef3 (Capsicum chinense defensin 3). The antifungal activity of these fractions was analyzed against yeasts of the genus Candida. At 200 μg/mL, fraction F1 inhibited the growth of C. tropicalis by 26%, fraction F2 inhibited 35% of the growth of C. buinensis, and fraction F3 inhibited all tested yeasts, exhibiting greater inhibition activity on the growth of the yeast C. albicans (86%) followed by C. buinensis (69%) and C. tropicalis (21%). Fractions F1 and F2 promoted membrane permeabilization of all tested yeasts and increased the endogenous induction of reactive oxygen species (ROS) in C. buinensis and C. tropicalis, respectively. We also observed that fraction F3 at a concentration of 50 µg/mL inhibited the α-amylase activities of Tenebrio molitor larvae by 96% and human salivary by 100%. Thus, our results show that fraction F3, which contains CcDef3, is a very promising protein fraction because it has antifungal potential and is able to inhibit the activity of different α-amylase enzymes.Keywords
Funding Information
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (E-26/203.090/2016, E-26/202.132/2015, E-26/202.330/2019)
This publication has 53 references indexed in Scilit:
- Capsicum annuum L. trypsin inhibitor as a template scaffold for new drug development against pathogenic yeastAntonie van Leeuwenhoek, 2011
- Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel α‐amylase inhibitory propertiesPhysiologia Plantarum, 2011
- Armed and Dangerous2010
- In-gel digestion for mass spectrometric characterization of proteins and proteomesNature Protocols, 2006
- Oxidative stress and cell cycle checkpoint function11Both Drs. Paules and Kaufmann received their doctoral degrees in Experimental Pathology from the University of North Carolina at Chapel Hill School of Medicine (in 1984 and 1979, respectively).Free Radical Biology and Medicine, 2000
- Fruit-Specific Expression of a Defensin-Type Gene Family in Bell Pepper (Upregulation during Ripening and upon Wounding)Plant Physiology, 1996
- α-amylase inhibitors from wheat: Amino acid sequences and patterns of inhibition of insect and human α-amylasesInsect Biochemistry and Molecular Biology, 1996
- An automated quantitative assay for fungal growth inhibitionFEMS Microbiology Letters, 1990
- Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDaAnalytical Biochemistry, 1987
- Measurement of protein using bicinchoninic acidAnalytical Biochemistry, 1985