Abstract
About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.
Highlights
According to the global consumption ranking, Phaseolus vulgaris (PV) is the third most important legume, preceded only by soybean (Glycine max) and peanut (Arachis hypogea)
Based on the UniProt database, it was possible to identify 35 peptides present in the PV3 extract that corresponded to 20 proteins from Phaseolus vulgaris
The method of choice for obtaining PV3 did not use techniques intended for chemical or physical digestion of proteins, i.e., the 35 peptides identified are naturally occurring in the grains
Summary
According to the global consumption ranking, Phaseolus vulgaris (PV) is the third most important legume, preceded only by soybean (Glycine max) and peanut (Arachis hypogea). Insolubilization of pectic substances by the phytase enzyme is a widely accepted mechanistic hypothesis [3]; Effects of bean peptides on endothelium there are other enzymatic reactions that may be involved in the hardening process Among these additional hypotheses, the following can be highlighted: formation of insoluble pectates; increase in the content of condensed tannins in the husk and migration of soluble tannins to the grain cotyledons; complex formation between tannins and medium lamella macromolecules; denaturation and/or protein association; complex formation between polyphenols, proteins, and pectins; cell wall lignification; oxidation and/or lipid polymerization [4]. This amount of disposable beans may still be used as a source of nutraceutical components [6,7], avoiding food wasting and obtaining some therapeutic benefits
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