Identification and in vitro Characterization of Novel Antidiabetic Peptides Released Enzymatically from Peanut Protein.

Bioactive peptides derived from proteins found in various foods provide significant health benefits, including regulating blood sugar levels by inhibiting carbohydrate-hydrolyzing enzymes. Hydrolysates of peanut protein were prepared using alcalase (AH) or trypsin (TH) to generate antidiabetic peptides with high activity against α-amylase (IC50 of 6.46 and 5.71 mg/mL) and α-glucosidase (IC50 of 6.30 and 5.57 mg/mL), as well as antiradical activity to scavenge DPPH• (IC50 of 4.18 and 3.12 mg/mL) and ABTS•+ (IC50 of 2.87 and 2.56 mg/mL), respectively. The bioactivities of hydrolysates were greatest in the ultrafiltration-generated F3 fraction (< 3 kDa). The most active fraction was TH-F3, which was purified by gel filtration chromatography to generate sub-fractions (SF). With IC50 values of 1.05 and 0.69 mg/mL, the F3-SF8 fraction was the most effective at inhibiting the activity of α-amylase and α-glucosidase, respectively. This fraction was further purified using RP-HPLC to generate sub-subfractions (SSF), the most active of which were F3-SF8-SSF9 and SSF10. The peptide sequences F3-SF8-SSF9 and SSF10 were determined using LC-MS/MS. Two novel antidiabetic peptides with the potential to inhibit α-amylase and α-glucosidase were identified, with the sequences Asp-Trp-Arg (476.22 Da, IC50 of 0.78, and 0.35 mg/mL) and Phe-Tyr (329.15 Da, IC50 of 0.91, and 0.41 mg/mL). These results suggest that peptides derived from peanut protein are attractive natural ingredients for diabetes management applications.

Proximate composition, nutritional evaluation and functional properties of a promising food: Arabian wax Cissus (Cissus rotundifolia Forssk) leaves.

Cissus rotundifolia is a wild plant, extensively used during scarcity and famine; however, the information about its chemical and nutritional properties still limited. In this work, C. rotundifolia was evaluated for its chemical, nutritional and functional properties. The results revealed that C. rotundifolia mainly contained carbohydrates (72.54%), proteins (12.16%), ash (12.53%), dietary fiber (14.10%), in addition to adequate amounts of essential amino acids and minerals. Fructose, glucose, galactose, and arabinose were the major monosaccharides with a percentage of 23.32, 13.60, 1.24, and 0.25 g/100 g DW, respectively. Furthermore, it was found that C. rotundifolia contained important vitamins, including thiamin (5.37 mg/100 g DW), riboflavin (1.19 mg/100 g DW), pyridoxine (0.46 mg/100 g DW) and folic acid (0.20 mg/100 g DW). The findings of functional properties revealed good water and oil absorption capacities of 2.74 and 1.63 g/g, respectively. Foaming capacity and water solubility index were 14 and 18.74%, respectively. From these results, it can be stated that C. rotundifolia has high nutritional values, which could be used widely in food applications.

Response Surface Modeling and Optimization of Enzymolysis Parameters for the In Vitro Antidiabetic Activities of Peanut Protein Hydrolysates Prepared Using Two Proteases.

Optimization of the enzymolysis process for preparing peanut protein hydrolysates using alcalase and trypsin was performed by employing the central composite design (CCD) of response surface methodology (RSM). The independent variables were solid-to-liquid ratio (S/L), enzyme-to-substrate ratio (E/S), pH, and reaction temperature, while the response variables were degree of hydrolysate (DH), α-amylase, and α-glucosidase inhibitory activity. The highest DH (22.84% and 14.63%), α-amylase inhibition (56.78% and 40.80%), and α-glucosidase inhibition (86.37% and 86.51%) were obtained under optimal conditions, which were S/L of 1:26.22 and 1:30 w/v, E/S of 6% and 5.67%, pH of 8.41 and 8.56, and temperature of 56.18 °C and 58.75 °C at 3 h using alcalase (AH) and trypsin (TH), respectively. Molecular weight distributions of peanut protein hydrolysates were characterized by SDS-PAGE, which were mostly ˂10 kDa for both hydrolysates. Lyophilized AH and TH had IC50 values of 6.77 and 5.86 mg/mL for α-amylase inhibitory activity, and 6.28 and 5.64 mg/mL for α-glucosidase inhibitory activity. The IC50 of AH and TH against DPPH radical was achieved at 4.10 and 3.20 mg/mL and against ABTS radical at 2.71 and 2.32 mg/mL, respectively. The obtained hydrolysates with antidiabetic activity could be utilized as natural alternatives to synthetic antidiabetics, particularly in food and pharmaceutical products.

Structural and physicochemical characteristics of lyophilized Chinese sturgeon protein hydrolysates prepared by using two different enzymes.

The structural and physicochemical characteristics of protein hydrolysates prepared from Chinese sturgeon through the enzymatic hydrolysis process were evaluated. Two different enzymes including papain and alcalase 2.4L were used in the hydrolysis process. The papain enzyme significantly increased the degree of hydrolysis (20.62%) and decreased the ζ-potential (12.4 ± 1.31 mV) as compared to the alcalase enzyme, which represented 15.55% and 15.53 ± 0.77 mV, respectively. Alcalase 2.4L hydrolysate exhibited smaller particle size (822.047 ± 61.26 nm) than papain hydrolysate (1425.39 ± 44.82 nm). Hydrolysis by papain and alcalase 2.4L enzymes decreased the molecular weights (MW ≤ 1,000 Da) to 98.27% and 86.84%, respectively. The surface hydrophobicity and turbidity of the hydrolysates significantly affected by enzyme type and protein concentrations. By using the X-ray diffraction analysis, the papain hydrolysate showed a higher relative crystallinity degree (30.33%) than alcalase 2.4L hydrolysate (29.40%), whereas the Fourier transform infrared spectroscopy showed more clearly peaks for the amide bands of alcalase hydrolysate. The thermal properties also affected by enzymatic hydrolysis conditions, since the melting temperatures were 159.17 and 149.58 °C, whereas the rate of mass loss was 67.04% and 62.8%, for papain and alcalase hydrolysates, respectively. PRACTICAL APPLICATION: The enzymatic hydrolysis process of proteins is employed to obtain the nutritionally and functionality important peptides that result during the preparation of fish protein hydrolysate. The objective of this study was to investigate the structural and physicochemical characteristics of protein hydrolysate prepared from Chinese sturgeon. This study showed that these characteristics were affected by enzymatic hydrolysis conditions especially enzyme type. The finding of this study may be useful in terms of providing new information on the properties of the protein hydrolysate and the structural changes resulting from controlled enzymatic hydrolysis conditions. Overall, these conditions could potentially alter the secondary structure of the protein hydrolysates or peptides and enhance their functional properties.

Effects of ultrasonic, microwave, and combined ultrasonic-microwave pretreatments on the enzymatic hydrolysis process and protein hydrolysate properties obtained from Chinese sturgeon (Acipenser sinensis).

Degree of hydrolysis (DH), yield, amino acid profile, protein solubility, and antioxidant activity of Chinese sturgeon protein hydrolysates, as influenced by thermal pretreatment, ultrasonic (US), microwave (MW), and combined US-microwave pretreatments were investigated. Initially, the samples were subjected to thermal pretreatments in order to measure their effect on DH, which increased at 55°C. The DH recorded 7.63, 5.55, and 6.02% for US, MW, and combined US-MW pretreatment (US + MW), respectively, at the optimal time (8 min). The enzymatic hydrolysis (EN) of pretreated samples increased the DH to 19.41, 14.18, and 16.91% for US + EN, MW + EN, and US + MW + EN, respectively. The US + EN treatment was most effective for obtaining higher DH and yield, which were 19.41% and 18.62%, respectively. The use of US + EN also resulted in an increase in the percentage of molecular weights (≤1,000 Da), amino acid content and protein solubility, which reached 89.24, 80.08, and 98.58%, respectively. While, US + MW + EN pretreatment has achieved the highest antioxidant activities by IC50 of 1,1-diphenyl-2-picrylhydrazyl and 2,2-Azinobis (3-ehtylbenzothiazoli- 6-sulfnic acid), which were 3.01 and 1.85 mg/ml, respectively, in addition to the reducing power assay, which was 0.528 at a protein concentration of 5 mg/ml. Therefore, US and combined US-MW techniques can play a promising role in the production of protein hydrolysates and the improvement of their antioxidant properties. PRACTICAL APPLICATIONS: Nowadays, interest in Chinese sturgeon production has increased as a promising source of protein and antioxidant peptides. The optimal thermal pretreatment can be used to enhance the degree of hydrolysis. The results indicated that the use of ultrasound as a pretreatment enhanced the degree of hydrolysis, which could be useful in the preparation of protein hydrolysate with higher yields. The use of combined US-MW significantly improved the antioxidant properties of the protein hydrolysate. The combined US-MW technique is a novel method for obtaining valuable peptides and protein hydrolysates that can be applied as antioxidant constituents in the food products.