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Alkaline fermentation and enzymatic hydrolysis of lentil favors the release of bioactive compounds with protective effects against oxidative stress

AuthorsBautista-Expósito, Sara; Peñas, Elena ; Silván, José Manuel ; Dueñas, Montserrat; Frías, Juana ; Martínez Villaluenga, Cristina
Issue Date2017
CitationEuroFoodChem XIX Conference (2017)
AbstractOxidative stress (excessive levels of radical oxygen species [ROS]) is involved in the initiation and progression of many chronic diseases. Dietary antioxidant compounds have a protective role against oxidative damage through different mechanisms. Legumes are sources of antioxidant peptides and phenolic compounds that must be released from food matrix components to become bioactive. Fermentation or enzymatic hydrolysis are used to release these bioactive compounds; however, combination of both treatments in alkaline conditions aiming at producing ingredients with antioxidant properties has not been explored yet. The objective of this study was to evaluate and compare the antioxidant activity of lentil soluble fractions obtained by three processing methods: fermentation with Lactobacillus plantarum (LP), enzymatic hydrolysis with Savinase® (S) and combination of the aforementioned treatments (LP+S). All treatments were performed at 37ºC, pH 8.5 for 15 h. Soluble fraction of raw lentil flour was used as control (C). Peroxyl radical scavenging activity of lentil soluble fractions was determined in vitro using oxygen radical absorbance capacity (ORAC) assay. The protective effect of lentil soluble fractions against oxidative stress was determined in RAW 264.7 macrophages cell line exposed to tert-butylhydroperoxide (t-BOOH). Cell viability and intracellular ROS levels were measured using tetrazolium dye and dichlorofluorescin probe, respectively. LP+S showed higher ORAC (508 mM Trolox equivalents [TE]/g) than LP (260 mM TE/g), S (384 mM TE/g) and C (302 mM TE/g). Cells treated with 100 µg/mL of LP, S or LP+S for 20 h showed similar viability to non-treated cells, indicating no cellular stress or damage. Under extreme oxidative conditions (2.5 mM t-BOOH for 3 h), pretreatment of macrophages with 100 µg/mL of LP, S and LP+S for 20 h greatly reduced intracellular ROS generation (50, 40, and 40% respectively) while C pretreatment was less effective (20%). Fractionation of LP+S by size exclusion chromatography was performed to further identify the main bioactive compounds. A total of 6 fractions were collected and tested for their antioxidant activity. Fraction II containing 560 mg/g of peptides and 50 mg/g of phenolic compounds was the most active reducing ROS in t-BOOH induced macrophages (53%) while also showed a high ORAC value (497 mM TE/g). HPLC-ESI-MS/MS analysis of fraction II allowed the identification of 31 peptides from convicilin, lectin and allergen Lenc1 and 7 phenolic compounds, mainly flavonoids, being kaempferol rutinoside-hexoside the flavonoid that presented higher concentration.. In conclusion, lentil flour processing by combination of fermentation and enzymatic hydrolysis in alkaline conditions may be useful to produce functional ingredients containing bioactive peptides and bound phenolics with enhanced peroxyl radical scavenging activity and protective effects against oxidative stress.
DescriptionResumen del póster presentado a la EuroFoodChem XIX Conference, celebrada en Budapest (Hungria) del 4 al 6 de octubre de 2017.
Appears in Collections:(ICTAN) Comunicaciones congresos
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