3 research outputs found
Isolation and identification of cholesterol esterase and pancreatic lipase inhibitory peptides from brewerâs spent grain by consecutive chromatography and mass spectrometry
The isolation and identification of cholesterol esterase (CE) and pancreatic lipase (PL) inhibitory peptides obtained from the protein hydrolysate of brewer's spent grain (BSG) was performed. BSG peptides were fractionated and purified sequentially by anion exchange, gel filtration (FPLC), and reversed phase high-performance liquid chromatography (RP-HPLC). The fractions obtained from each chromatographic step were collected and the in vitro enzyme inhibitory activity was evaluated. The chromatographic purification process increased the in vitro activities. The most active fractions were evaluated using MALDI-TOF tandem mass spectrometry, which identified three peptides: a peptide with the highest CE inhibition capacity (WNIHMEHQDLTTME) and two peptides with PL inhibition capacity (DFGIASF and LAAVEALSTNG). These three peptides showed hydrophobic and acidic amino acid residues (Asp and Glu) and/or their amines (Asn and Gln), which could be a common feature among lipid-lowering peptides related to CE and PL enzyme inhibition. The in silico studies showed that the three peptides had high hydrophobicity and were susceptible to enzymatic hydrolysis performed by trypsin, pepsin, and pancreatin. The BSG byproduct was a good source of CE and PL inhibitory peptides, thus adding value to this byproduct of the beer industry. This is the first report to demonstrate that BSG peptides can inhibit CE and PL enzymes. This journal isFil: GarzĂłn, Antonela Guadalupe. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; ArgentinaFil: Cian, RaĂșl Esteban. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; ArgentinaFil: Aquino, MarilĂn EstefanĂa. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; ArgentinaFil: Drago, Silvina Rosa. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; Argentin
Anti-diabetic properties of brewerâs spent yeast peptides. In vitro, in silico and ex vivo study after simulated gastrointestinal digestion
Brewerâs spent yeast (BSY) hydrolysates are a source of antidiabetic peptides. Nevertheless, the impact of
in vitro gastrointestinal digestion of BSY derived peptides on diabetes has not been assessed. In this study,
two BSY hydrolysates were obtained (H1 and H2) using ÎČ-glucanase and alkaline protease, with either 1 h
or 2 h hydrolysis time for H1 and H2, respectively. These hydrolysates were then subjected to simulated
gastrointestinal digestion (SGID), obtaining dialysates D1 and D2, respectively. BSY hydrolysates inhibited
the activity of α-glucosidase and dipeptidyl peptidase IV (DPP-IV) enzymes. Moreover, although D2 was
inactive against these enzymes, D1 IC50 value was lower than those found for the hydrolysates.
Interestingly, after electrophoretic separation, D1 mannose-linked peptides showed the highest
α-glucosidase inhibitory activity, while non-glycosylated peptides had the highest DPP-IV inhibitory
activity. Kinetic analyses showed a non-competitive mechanism in both cases. After peptide identification,
GILFVGSGVSGGEEGAR and IINEPTAAAIAYGLDK showed the highest in silico anti-diabetic activities
among mannose-linked and non-glycosylated peptides, respectively (AntiDMPpred score: 0.70 and 0.77).
Molecular docking also indicated that these peptides act as non-competitive inhibitors. Finally, an ex vivo
model of mouse jejunum organoids was used to study the effect of D1 on the expression of intestinal epithelial
genes related to diabetes. The reduction of the expression of genes that codify lactase, sucraseisomaltase
and glucose transporter 2 was observed, as well as an increase in the expression of Gip
(glucose-dependent insulinotropic peptide) and Glp1 (glucagon-like peptide 1). This is the first report to
evaluate the anti-diabetic effect of BSY peptides in mouse jejunum organoidsAgencia Nacional de PromociĂłn CientĂfica y TecnolĂłgica (ANPCyT) - Argentina (Project PICT-2020-Serie A-1985)Agencia Santafesina de Ciencia TecnologĂa e InnovaciĂłn â Argentina (Res. 132/23-Project PEICID-2022-133)Ministerio de Ciencia e InnovaciĂłn MICINN grant no. PID2020-112768RB-I00Instituto de Salud Carlos III, grant no. PI21/00952Junta de AndalucĂa and FEDER Grants A-AGR-468-UGR20 and P20-0069
Anti-diabetic properties of brewer's spent yeast peptides: In vitro, in silico and ex vivo study after simulated gastrointestinal digestion
BrewerÂŽs spent yeast (BSY) hydrolysates are a source of antidiabetic peptides. Nevertheless, the impact of in vitro gastrointestinal digestion of BSY derived peptides on diabetes has not been assessed. In this study, two BSY hydrolysates were obtained (H1 and H2) using ÎČ-glucanase and alkaline protease, with either 1 h or 2 h hydrolysis time for H1 and H2, respectively. These hydrolysates were then subjected to simulated gastrointestinal digestion (SGID), obtaining dialysates D1 and D2, respectively. BSY hydrolysates inhibited the activity of α-glucosidase and dipeptidyl peptidase IV (DPP-IV) enzymes. Moreover, although D2 was inactive against these enzymes, D1 IC50 value was lower than those found for the hydrolysates. Interestingly, after electrophoretic separation, D1 mannose-linked peptides showed the highest α-glucosidase inhibitory activity, while non-glycosylated peptides had the highest DPP-IV inhibitory activity. Kinetic analyses showed a non-competitive mechanism in both cases. After peptide identification, GILFVGSGVSGGEEGAR and IINEPTAAAIAYGLDK showed the highest in silico anti-diabetic activities among mannose-linked and non-glycosylated peptides, respectively (AntiDMPpred score: 0.70 and 0.77). Molecular docking also indicated that these peptides act as non-competitive inhibitors. Finally, an ex vivo model of mouse jejunum organoids was used to study the effect of D1 on the expression of intestinal epithelial genes related to diabetes. The reduction of the expression of genes that codify lactase, sucrase-isomaltase and glucose transporter 2 was observed, as well as an increase in the expression of Gip (glucose-dependent insulinotropic peptide) and Glp1 (glucagon-like peptide 1). This is the first report to evaluate the anti-diabetic effect of BSY peptides in mouse jejunum organoids.Fil: Aquino, MarilĂn EstefanĂa. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; ArgentinaFil: Drago, Silvina Rosa. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; ArgentinaFil: SĂĄnchez de Medina, FermĂn. Universidad de Granada. Facultad de Farmacia.; EspañaFil: MartĂnez Augustin, Olga. Universidad de Granada. Facultad de Farmacia.; EspañaFil: Cian, RaĂșl Esteban. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de IngenierĂa QuĂmica. Instituto de TecnologĂa de los Alimentos; Argentin