Millet bran protein hydrolysates derived peptides-zinc chelate: Structural characterization, security prediction in silico, zinc transport capacity and stability against different food processing conditions.

A novel peptide Ser-Asp-Asp-Val-Leu (SDDVL) of excellent zinc-chelating capacity (13.77 mg/g) was identified in millet bran protein hydrolysates. In silico prediction demonstrated that SDDVL had no potential toxicity. The results of structural characterization demonstrated that both amino group and carboxyl group of SDDVL were the primary zinc-chelating sites. Moreover, SDDVL-zinc chelate showed higher stability (p < 0.05) than ZnSO4 and zinc gluconate under different processing conditions including most pasteurization conditions, heating at 100°C for 10-50 min, various pH values (8.0-10.0), treatment of glucose (4-8 g/100 g) or NaCl (1-4 g/100 g), and simulated gastrointestinal digestion. In addition, SDDVL-zinc chelate showed higher zinc transport capacity than ZnSO4 and zinc gluconate in Caco-2 cells (p < 0.05). These results suggested that millet bran peptide had a positive effect on the gastrointestinal stability and bioavailability of Zn, and SDDVL-zinc chelate could be used as ingredient of zinc supplements. PRACTICAL APPLICATION: The current study provided a practical method to identify peptides of excellent zinc-chelating capacity from millet bran protein hydrolysates. This study demonstrated that in silico prediction assisted with suitable database was a fast, practical, and economic way to evaluate the security and to analysis the physicochemical properties of novel peptides. Moreover, it provided an efficient method to assess the stability of peptide-zinc chelate under different food processing conditions, which was the theoretical basis for utilization of peptide as ingredient of zinc fortifications.

The influences of acetylation, hydroxypropylation, enzymatic hydrolysis and crosslinking on improved adsorption capacities and in vitro hypoglycemic properties of millet bran dietary fibre.

The effects of acetylation, hydroxypropylation, cellulase hydrolysis and crosslinking on adsorption capacities and in vitro hypoglycemic activities of millet bran dietary fibre (MBDF) were studied. The results demonstrated that both acetylation and hydroxypropylation improved water swelling ability of MBDF, and adsorption capacities of cholesterol, cholate and copper ion on MBDF. Acetylation and hydroxypropylation also enhanced α-glucosidase and α-amylase inhibition activities, glucose-binding ability and glucose diffusion retardation index (GDRI) of MBDF. Acetylated MBDF showed the highest cholate (77.31 mg/g) and cholesterol (13.97 mg/g) adsorption capacities. The crosslinking improved adsorption of cholate, cholesterol, copper ion (25.64 mg/g) and nitrite ion (181.59 µg/g) on MBDF; but reduced α-amylase inhibition activity (p < 0.05). Moreover, cellulase hydrolyzed MBDF exhibited the highest GDRI (39.60%) and α-amylase inhibition activity (34.53%), but the lowest oil and cholate adsorption capacities. The results suggest that the modified MBDFs can be used as an ingredient of hypoglycemic foods.

Effect of four modification methods on adsorption capacities and in vitro hypoglycemic properties of millet bran dietary fibre.

To improve the adsorption capacities and hypoglycemic properties of millet bran dietary fibre (MBDF), four methods including acrylate-grafting, carboxymethylation, heat assisted with cellulase hydrolysis, and enzymatic hydrolysis combined with acrylate-grafting were used. The results demonstrated that all carboxymethylation, acrylate-grafting, and enzymatic hydrolysis combined with acrylate-grafting improved soluble dietary fibre content, water swelling ability and α-amylase-inhibition activity of MBDF. They also increased oil, cholesterol, sodium cholate, copper ion and nitrite ion adsorption capacities of MBDF. But carboxymethylation, acrylate-grafting and enzymatic hydrolysis combined with acrylate-grafting decreased polyphenol content, glucose-binding ability and glucose dialysis retardation index of MBDF (p < 0.05). The heat assisted with cellulase hydrolysis increased soluble dietary fibre content, polyphenol content, sodium cholate-adsorption capacity, and hypoglycemic properties of MBDF including glucose-binding ability, glucose dialysis retardation index and α-amylase-inhibition activity; but reduced adsorption capacity of MBDF on cholesterol and copper ion (p < 0.05). Changes in structure of MBDF caused by these modification methods were proved by the results of scanning electron microscopy and Fourier-transformed infrared spectroscopy analysis. These results highlight potential applications of these modified MBDFs as ingredients of hypolipidemic and hypoglycemic foods, or scavenger of nitrite and copper ion.

Effects of carboxymethylation, hydroxypropylation and dual enzyme hydrolysis combination with heating on physicochemical and functional properties and antioxidant activity of coconut cake dietary fibre.

The effects of carboxymethylation, hydroxypropylation and dual enzyme hydrolysis combined with heating on some physicochemical and functional properties, and antioxidant activity of coconut cake dietary fibre (CCDF) were studied. Results showed that both the hydroxypropylation and carboxymethylation could effectively improve (p < 0.05) the water retention capacity (WRC), oil retention capacity (ORC), viscosity, α-amylase inhibition activity (α-AAIR), glucose dialysis retardation index (GDRI), cation-exchange capacity, emulsifying capacity index (ECI) and bile adsorption capacity (BAC) of CCDF. Moreover, the cellulase and hemicellulase hydrolysis combination with heating significantly enhanced (p < 0.05) the soluble dietary fibre content, WRC, emulsion stability, GDRI, α-AAIR and BAC of CCDF; but caused decrease in ORC and browning of color. In addition, improvement of total phenol content, Fe2+ chelating ability, ABTS+· and O2-· scavenging activity were obtained in carboxymethylaticted CCDF. These effects were mainly attributed to the composition and structural modifications as evident from SEM, FT-IR and XRD analysis.

Isolation of novel ACE-inhibitory peptide from naked oat globulin hydrolysates in silico approach: Molecular docking, in vivo antihypertension and effects on renin and intracellular endothelin-1.

Naked oat globulin was hydrolyzed by alcalase, flavourzyme, pepsin, and trypsin in sequence. The hydrolysates (NOGH) were purified using gel chromatography, reversed-phase high performance liquid chromatography (RP-HPLC). Finally, fraction D7d with the highest ACE-inhibitory was subjected to liquid chromatography-mass spectrometry analysis and 14 peptides were identified. Of which, peptide SSYYPFK (890.4 Da) was chose to synthesize based on in silico analysis. The SSYYPFK demonstrated high ACE-inhibitory activity (IC50 : 91.82 µM) with competitive inhibition mode, and could effectively (P < 0.05) lower the systolic blood pressure and diastolic pressure of spontaneously hypertensive rats at the concentration of 100 to 150 mg/kg body weight. Molecular docking simulation demonstrated that SSYYPFK could bind with the active site S1 of ACE via short hydrogen bonds. It could remain the ACE-inhibitory activity after simulated gastrointestinal hydrolysis. Moreover, SSYYPFK showed acceptable renin and endothelin-1 suppressing capacity (47.59% and 27.88% at 1.5 mg/mL, respectively). These results indicated that SSYYPFK may have similar antihypertensive mechanism with captopril, and could be develop to natural antihypertensive products. PRACTICAL APPLICATION: One novel ACE-inhibitory peptide SSYYPFK (890.4 Da) was identified from naked oat globulin hydrolysates. It exhibited relatively high renin and intracellular endothelin-1 suppressing capacity, and could effectively (P < 0.05) lower the systolic blood pressure and diastolic pressure of spontaneously hypertensive rats. This peptide could be used as natural and safe nutraceuticals and/or functional ingredients.

Isolation of Novel ACE-Inhibitory and Antioxidant Peptides from Quinoa Bran Albumin Assisted with an In Silico Approach: Characterization, In Vivo Antihypertension, and Molecular Docking.

Albumin is the major fraction of quinoa protein that is characterized as having high nutritional value. However, until now, scant information is available on the bioactivity of quinoa albumin or its hydrolysates. To promote its usage, we extracted albumin in this study from quinoa bran assisted with cellulase and hemicellulose, and hydrolyzed it by alcalase and trypsin to produce bioactive peptides. The hydrolysates (QBAH) were purified by gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC), followed by identification using liquid chromatography-mass spectrometry (LC-MS/MS). Furthermore, based on in silico analysis, one angiotensin-I converting enzyme (ACE)-inhibitory and antioxidant peptide, RGQVIYVL (946.6 Da), and two antioxidant peptides, ASPKPSSA (743.8 Da), and QFLLAGR (803.5 Da), from QBAH were synthesized. RGQVIYVL showed a high ACE-inhibitory activity (IC50 = 38.16 µM) with competitive mode of inhibition, and showed significant antihypertensive effect in spontaneously hypertensive rats at a concentration of 100-150 mg/kg body weight (bw). Molecular docking simulation showed that it could interact with the active ACE site via hydrogen bonds with high binding power. Moreover, RGQVIYVL, ASPKPSSA, and QFLLAGR all demonstrated high ·OH scavenging activity (IC50 = 61.69-117.46 µM), ABTS+ scavenging activity (58.29-74.28%) and Fe2+ chelating ability (32.54-82.48% at 0.5 mg/mL). They could also retain activity after gastrointestinal enzyme digestion. These results indicate that quinoa albumin is a potential source of bioactive peptides possessing antioxidant and ACE-inhibitory activities.