RESUMEN
Bioactivity of a snack-bar, chocolate-drink and yogurt fortified with brewers' spent grain (BSG) phenolic extracts (P2 or B2) or protein hydrolysates (barley protein hydrolysate (BPH), BPH < 3 kDa, BPH < 5 kDa, BPH > 5 kDa) was measured following gastrointestinal in vitro digestion. Concentrations of 0.5 and 0.1% (v/v) digestates were chosen for addition to Caco-2 and Jurkat T cells, respectively. Yogurt and B2 digestate protected against H2O2-induced DNA damage in Caco-2 cells (p < 0.05), by the comet assay. Snack-bar digestates possessed significant (p < 0.05) immunomodulatory effects, measured by ELISA in concanavalin-A stimulated Jurkat T cells. Addition of BPH enhanced (p < 0.05) the IFN-γ reducing capacity of the snack-bar while addition of BPH < 3 and < 5 kDa reduced IL-2 production to a greater extent than unfortified yogurt (p < 0.05). Selected BSG components can enhance the antioxidant and anti-inflammatory potential of foods.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Grano Comestible/química , Alimentos Fortificados/análisis , Hordeum/química , Fenoles/farmacología , Hidrolisados de Proteína/farmacología , Células CACO-2 , Concanavalina A/inmunología , Daño del ADN/efectos de los fármacos , Dieta , Alimentos Formulados , Tracto Gastrointestinal , Humanos , Peróxido de Hidrógeno , Factores Inmunológicos , Técnicas In Vitro , Interferón gamma/metabolismo , Interleucina-2/metabolismo , Células Jurkat , Extractos Vegetales/farmacología , Bocadillos , YogurRESUMEN
Brewers' spent grain (BSG) protein rich fraction was previously hydrolysed using Alcalase (U) and three additional fractions were prepared by membrane fractionation; a 5-kDa retentate (U > 5), a 5-kDa permeate (U < 5) and a 3-kDa permeate (U < 3). In the present study, these fractions were added to milk, subjected to simulated gastrointestinal digestion (SGID) and their anti-inflammatory potential was investigated. The digestates caused a significant reduction (p < 0.05) in interleukin-6 (IL-6) production in Concanavalin-A (ConA)-stimulated Jurkat T cells. The samples did not significantly alter the production of IL-6 in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. IL-2 and interferon-γ (IFN-γ) production in stimulated Jurkat T cells and IL-1ß and tumor necrosis factor-α (TNF-α) production in stimulated RAW 264.7 cells were not affected in the presence of the digestates. Results show that a SGID milk product supplemented with BSG hydrolysate and its associated ultrafiltered fractions can confer anti-inflammatory effects in Jurkat T cells.
Asunto(s)
Grano Comestible/química , Tracto Gastrointestinal/metabolismo , Factores Inmunológicos/farmacología , Leche/química , Proteínas de Plantas/química , Hidrolisados de Proteína/química , Animales , Proliferación Celular , Citocinas/metabolismo , Suplementos Dietéticos , Digestión , Alimentos Formulados , Alimentos Fortificados/análisis , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Factores Inmunológicos/química , Células Jurkat , RatonesRESUMEN
BACKGROUND: Large quantities of brewers' spent grain (BSG), a co-product of the brewing industry, are produced annually. BSG contains hydroxycinnamic acids, and phenolic-rich extracts from BSG have previously demonstrated the ability to protect against oxidant-induced DNA damage. The present study investigated the anti-inflammatory potential of eight phenolic extracts from BSG: four pale (P1-P4) and four black (B1-B4) extracts. RESULTS: BSG extracts were more cytotoxic in Jurkat T than U937 cells, with lower IC50 values in Jurkat T cells, measured using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Pale BSG extracts P2 and P3 showed the greatest anti-inflammatory potential, significantly (P < 0.05) reducing interleukin-2 (IL-2), interleukin-4 (IL-4, P2 only), interleukin-10 (IL-10) and interferon-γ (IFN-γ) production. In addition, extracts P1-P3 and B2-B4 showed significant (P < 0.05) antioxidant effects, determined by the cellular antioxidant activity assays superoxide dismutase, catalase and glutathione content (GSH). CONCLUSION: Phenolic extracts from BSG, particularly the pale BSG extracts, have the ability to reduce a stimulated cytokine production and may also protect against cellular oxidative stress. Results of the present study highlight the potential of BSG phenolic extracts to act as functional food ingredients, providing an alternative use and improving the value of this brewing industry co-product.
Asunto(s)
Antioxidantes/metabolismo , Cerveza , Ácidos Cumáricos/metabolismo , Grano Comestible/química , Factores Inmunológicos/metabolismo , Residuos Industriales/análisis , Fenoles/metabolismo , Antiinflamatorios no Esteroideos/análisis , Antiinflamatorios no Esteroideos/economía , Antiinflamatorios no Esteroideos/aislamiento & purificación , Antiinflamatorios no Esteroideos/metabolismo , Antineoplásicos Fitogénicos/análisis , Antineoplásicos Fitogénicos/economía , Antineoplásicos Fitogénicos/aislamiento & purificación , Antineoplásicos Fitogénicos/metabolismo , Antioxidantes/análisis , Antioxidantes/economía , Antioxidantes/aislamiento & purificación , Cerveza/economía , Cerveza/microbiología , Línea Celular Tumoral , Ácidos Cumáricos/análisis , Ácidos Cumáricos/economía , Ácidos Cumáricos/aislamiento & purificación , Citocinas/antagonistas & inhibidores , Citocinas/metabolismo , Grano Comestible/economía , Alimentos Fortificados/análisis , Alimentos Fortificados/economía , Industria de Procesamiento de Alimentos/economía , Humanos , Factores Inmunológicos/análisis , Factores Inmunológicos/economía , Factores Inmunológicos/aislamiento & purificación , Residuos Industriales/economía , Irlanda , Leucemia de Células T/inmunología , Leucemia de Células T/metabolismo , Monocitos/enzimología , Monocitos/inmunología , Monocitos/metabolismo , Fenoles/análisis , Fenoles/economía , Fenoles/aislamiento & purificación , Pigmentación , Extractos Vegetales/química , Extractos Vegetales/economía , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/metabolismo , ReciclajeRESUMEN
The moderately thermophilic aerobic ascomycete Talaromyces emersonii secretes, under selected growth conditions, several ß-glucan hydrolases including an exo-1,3-ß-glucanase. This enzyme was purified to apparent homogeneity in order to characterise its biochemical properties and investigate hydrolysis of different ß-glucans, including laminaran, a 1,3-ß-glucan from brown algae. The native enzyme is monomeric with a molecular mass of ~40 kDa and a pI value of 4.3, and is active over broad ranges of pH and temperature, with optimum activity observed at pH 5.4 and 65 °C. At pH 5.0, the enzyme displays strict specificity for laminaran (apparent K(m) 1.66 mg mL⻹; V(max) 7.69 IU mL⻹) and laminari-oligosaccharides and did not yield activity against 1,4-ß-glucans, 1,3;1,4-ß-glucans or 4-nitrophenyl- and methylumbelliferyl-ß-D: -glucopyranosides. Analysis of hydrolysis products formed during time-course hydrolysis of laminaran by high-performance anion exchange chromatography with pulsed amperometric detection revealed a strict exo mode of action, with glucose being the sole reaction product even at the initial stages of hydrolysis. The T. emersonii exo-1,3-ß-glucanase was inhibited by glucono-δ-lactone (K(i) 1.25 mM) but at significantly higher concentrations than typically inhibitory for exo-glycosidases such as ß-glucosidase. 'De novo' sequence analysis of the purified enzyme suggests that it belongs to family GH5 of the glycosyl hydrolase superfamily. The results clearly show that the exo-1,3-ß-glucanase is yet another novel enzyme present in the ß-glucanolytic enzyme system of T. emersonii.
Asunto(s)
Glucano 1,3-beta-Glucosidasa/aislamiento & purificación , Glucano 1,3-beta-Glucosidasa/metabolismo , Talaromyces/enzimología , Cromatografía Líquida de Alta Presión , Inhibidores Enzimáticos/metabolismo , Estabilidad de Enzimas , Glucano 1,3-beta-Glucosidasa/química , Glucanos , Gluconatos/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Lactonas/metabolismo , Peso Molecular , Polisacáridos/metabolismo , Especificidad por Sustrato , TemperaturaRESUMEN
An alkaline extracted brewers' spent grain protein-enriched isolate (BSG-PI) was hydrolysed using Alcalase, Corolase PP, Flavourzyme and Promod 144MG, yielding Alc hydrolysate (H), CorH, FlavH and ProH, respectively. The degree of hydrolysis (DH) of the protein hydrolysates varied from 4.45% for ProH to 16.4% for CorH. The in vitro ACE inhibitory activity of the BSG-PI increased significantly following 15min incubations with Alcalase, Corolase PP and Flavourzyme. The 5kDa ultrafiltration permeates of FlavH and CorH resulted in lower ACE IC50 values than their respective hydrolysates. The bioactivity of the BSG-PI hydrolysates was retained after simulated gastrointestinal digestion (SGID) while SGID also resulted in the release of ACE inhibitory peptides from the BSG-PI and ProH. UPLC-MS/MS analysis resulted in the identification of 34 peptides. Of 12 synthesised peptides, IVY and ILDL were the most potent, having ACE IC50 values at 80.4±11.9 and 96.4±8.36µM, respectively.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Cromatografía Liquida/métodos , Grano Comestible/química , Proteínas de Plantas/química , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacología , Espectrometría de Masas en Tándem/métodos , Grano Comestible/metabolismo , Hidrólisis , Péptidos/química , Péptidos/farmacología , Proteínas de Plantas/metabolismo , Subtilisinas/químicaRESUMEN
The present study investigated the bioactivity of protein hydrolysates and fractionated hydrolysates prepared from brewers' spent grain (BSG) using proteases, including Alcalase 2.4L, Flavourzyme and Corolase PP. Hydrolysates were designated K-Y, including fractionated hydrolysates with molecular weight (m.w.) < 3, <5 and >5 kDa. Where computable, IC50 values were lower in U937 (1.38-9.78%) than Jurkat T cells (1.15-13.82%). Hydrolysates L, Q and R and fractionated hydrolysates of U and W (<3, <5, >5 kDa) significantly (P < 0.01) protected against hydrogen peroxide (H2O2)-induced reduction of superoxide dismutase (SOD) activity. A fractionated hydrolysate of W (<5 kDa) protected against H2O2-induced DNA damage, P < 0.01. Hydrolysates K, N, P, U, U > 5 kDa, V, V > 5 kDa, W, W > 5 kDa significantly (P < 0.05) reduced a concanavlin-A (con-A) stimulated production of interferon-γ (IFN-γ). In conclusion, BSG protein hydrolysates demonstrate bioactivity in vitro; lower m.w. hydrolysates (<3, <5 kDa) show greatest antioxidant activity and unfractionated or higher m.w. hydrolysates (>5 kDa) possess anti-inflammatory effects.
Asunto(s)
Antioxidantes/farmacología , Células/metabolismo , Concanavalina A/metabolismo , Grano Comestible/química , Peróxido de Hidrógeno/farmacología , Interferón gamma/metabolismo , Estrés Oxidativo/efectos de los fármacos , Hidrolisados de Proteína/farmacología , Antioxidantes/química , Proliferación Celular/efectos de los fármacos , Células/citología , Células/efectos de los fármacos , Humanos , Hidrolisados de Proteína/química , Células U937 , Residuos/análisisRESUMEN
Brewers' spent grain (BSG) is a low-value co-product of the brewing industry produced in large quantities annually. This paper reviews the existing evidence regarding the phenolic component of BSG, focusing on composition, extraction and biofunctions such as antioxidant, anti-atherogenic, anti-inflammatory and anti-carcinogenic activities. Furthermore, the incorporation of BSG in foodstuffs will be discussed, including the use of BSG as an animal feed supplement and the potential of BSG to be incorporated into foods for human consumption. BSG contains hydroxycinnamic acids including ferulic acid, p-coumaric acid and caffeic acid; which have shown bioactivity in the pure form (antioxidant, anti-inflammatory, anti-atherogenic and anti-cancer). Phenolic extracts from BSG have also shown antioxidant potential, by protecting against oxidant-induced DNA damage, possibly by Fe chelation. Studies show that BSG has many benefits when used as a supplement in animal feed, such as increasing milk yield, milkfat content and in providing essential dietary amino acids. The incorporation of BSG in human foods such as cookies and ready-to-eat snacks has resulted in increased protein and fibre contents of the products, where the changes in organoleptic properties are controllable. It can be concluded that the phenolic component of BSG has potential bioactive effects, which are worth pursuing given that the inclusion of BSG into human foodstuffs is viable and beneficial.
Asunto(s)
Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales/efectos de los fármacos , Dieta , Grano Comestible/química , Alimentos Funcionales , Fenoles/farmacología , Extractos Vegetales/farmacología , Animales , Suplementos Dietéticos , Fermentación , HumanosRESUMEN
The hydroxycinnamic acid (HA) content of starting barley for brewers' spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P<0.05) correlated with caffeic acid (R(2)=0.8309) and total HA (R(2)=0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P<0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P<0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.
Asunto(s)
Bebidas/análisis , Ácidos Cumáricos/análisis , Aditivos Alimentarios/análisis , Frutas/química , Hordeum/química , Fenoles/análisis , Extractos Vegetales/análisis , Antioxidantes/análisis , Cromatografía Líquida de Alta Presión , ResiduosRESUMEN
Brewers' spent grain (BSG), a by-product of the brewing industry, contains high amounts of phenolic acids, which have antioxidant effects. The present study examined the ability of BSG extracts to protect against the genotoxic effects of oxidants, hydrogen peroxide (H(2)O(2)), 3-morpholinosydnonimine hydrochloride (SIN-1), 4-nitroquinoline 1-oxide (4-NQO) and tert-butylhydroperoxide (t-BOOH) in U937 cells. Four pale (P1-P4) and four black (B1-B4) BSG extracts were investigated. U937 cells were pre-incubated with BSG extracts, exposed to the oxidants and the DNA damage was measured by the Comet assay. The black BSG extracts (B1-B4) significantly protected against H(2)O(2)-induced DNA damage. Extract B2, which had the highest phenol content, provided the greatest protection. Extracts P2, B2, B3 and B4 provided significant protection against SIN-1-induced DNA damage. None of the extracts protected against DNA damage induced by t-BOOH and 4-NQO. The DNA protective effects of the BSG phenolic extracts may be related to iron chelation.