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.

Microencapsulation of Bifidobacterium bifidum F-35 via modulation of emulsifying technique and its mechanical effects on the rheological stability of set-yogurt.

Microencapsulation of B. bifidum F-35 was carried out through emulsification technique in order to increase the microbial load while maintaining the rheological functions of set-yogurt. To produce single-layer (SL) microcapsules of whey protein, the pH was adjusted to 6.4 within Transglutaminase-induced gelation. Sodium alginate was processed as the external layer using calcium-induced gelation (pH 5.5) to produce the double-layer (DL) microcapsule. Scanning electron microscopy revealed that SL and DL microcapsules had sizes of 10 and 280 µm, respectively. The highest microbial load was clearly visible in the DL sample. According to texture profile analysis, the DL sample had the highest levels of gumminess, chewiness, and adhesiveness. The free sample outperformed the encapsulated samples in terms of springiness and cohesiveness. Although the SL sample had the highest viscosity, it produced a deformed gel when firmness was measured. In terms of firmness, the DL sample performed quite well. The viability of encapsulated B. bifidum F-35 in DL was higher than SL microcapsules during storage. Microencapsulation of B. bifidum F-35 with whey protein and sodium alginate is a promising technique that could improve the rheological properties of set-yogurt as a popular vehicle for bioactive ingredients.

Natural phospholipids: Occurrence, biosynthesis, separation, identification, and beneficial health aspects.

During the last years, phospholipids (PLs) have attracted great attention because of their crucial roles in providing nutritional values, technological and medical applications. There are considerable proofs that PLs have unique nutritional benefits on human health, such as reducing cholesterol absorption, improving liver functions, and decreasing the risk of cardiovascular diseases. PLs are the main structural lipid components of cell and organelle membranes in all living organisms, and therefore, they occur in all organisms and the derived food products. PLs are distinguished by the presence of a hydrophilic head and a hydrophobic tail, consequently they possess amphiphilic features. Due to their unique characteristics, the extraction, separation, and identification of PLs are critical issues to be concerned. This review is focused on the content of PLs classes in several sources (including milk, vegetable oils, egg yolk, and mitochondria). As well, it highlights PLs biosynthesis, and the methodologies applied for PLs extraction and separation, such as solvent extraction and solid-phase extraction. In addition, the determination and quantification of PLs classes by using thin layer chromatography, high-performance liquid chromatography coupled with different detectors, and nuclear magnetic resonance spectroscopy techniques.

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.

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.

Spray-dried novel structured lipids enriched with medium-and long-chain triacylglycerols encapsulated with different wall materials: Characterization and stability.

Novel structured lipids (SLs) enriched with medium-and long-chain triacylglycerols (MLCTs) were synthesized to combine the benefits of both arachidonic acid and medium-chain fatty acids; however, they are susceptible to oxidative degradation. In this work, the influences of the partial replacement of whey protein isolate (WPI) as the primary wall material by prebiotic carbohydrates, such as maltodextrin (MD) and inulin (IN) as the secondary wall materials on the physicochemical characteristics and oxidative stability of the spray-dried MLCTs-rich SLs microcapsules were investigated. The highest values of viscosity and zeta-potential were achieved by the WPI/IN (1:1) emulsions. Size distributions of all the emulsions were mono modal and became bimodal after microencapsulation process. The microcapsules prepared with WPI/IN (1:1) had the lowest lightness and the highest yellowness values. The partial replacement treatments increased the solubility and reduced the moisture content of the produced microcapsules. The partial replacement of WPI by IN significantly enhanced the encapsulation efficiency (89.10 ±â€¯1.03%), wettability properties (205 ±â€¯10.61 S), and decreased the incidence of surface oil on the microcapsules. The free oil content was noted as 5.73 ±â€¯0.05, 3.83 ±â€¯0.01, and 2.40 ±â€¯0.03% for the microcapsules produced using WPI, WPI/MD (1:1), and WPI/IN (1:1), respectively. Larger microcapsules and fairer flowing properties were achieved in the powders produced with only WPI. The partial replacement of WPI by IN provided the best oxidative stability of the microencapsulated MLCTs-rich SLs. The results revealed that MD and IN with WPI together, particularly IN proved to be a good substitute secondary wall material for spray-dried MLCTs-rich SLs, therefore suggesting its usefulness in functional food applications.

Synthesis of 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids by lipase-catalyzed acidolysis of microbial oil from Mortierella alpina.

Microbial oils (MOs) have gained widespread attention due to their functional lipids and health promoting properties. In this study, 1,3-dioleoyl-2-arachidonoylglycerol-rich structured lipids (SLs) were produced from MO and oleic acid (OA) via solvent-free acidolysis catalyzed by Lipozyme RM IM. Under the optimal conditions, the content of unsaturated fatty acids (UFAs) increased from 60.63 to 84.00%, while the saturated fatty acids (SFAs) content decreased from 39.37 to 16.00% at sn-1,3 positions in SLs. Compared with MO, arachidonic acid (ARA) content at the sn-2 position of SLs accounted for 49.71%, whereas OA was predominantly located at sn-1,3 positions (47.05%). Meanwhile, the most abundant triacylglycerol (TAG) species in SLs were (18:1-20:4-18:1), (20:4-20:4-18:1), (18:1-18:2-18:1), (18:1-18:2-18:0) and (24:0-20:4-18:1) with a relative content of 18.79%, 11.94%, 6.07%, 5.75% and 4.84%, respectively. Such novel SLs with improved functional properties enriched with UFAs are highly desirable and have the potential to be used in infant formula.

Identification of phospholipids classes and molecular species in different types of egg yolk by using UPLC-Q-TOF-MS.

Egg phospholipids (PLs) are currently the products of greatest commercial interest with major area of importance in various fields. Therefore, in this study, duck, hen and quail egg yolk PLs were isolated by solvent extraction with chilled acetone precipitation, and subsequently separated and identified by using ultra-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Egg PLs were separated on hydrophilic interaction liquid chromatography (HILIC) with ethylene bridged hybrid (BEH) column by gradient elution using acetonitrile/ammonium formate as a mobile phase, and detected by mass spectrometry (MS) under electrospray ionization in positive and negative ion mode. Structural characterizations of 57 molecular species of egg yolk PLs were identified based on MS/MS fragment ion information and elemental composition in MassLynx 4.1 software. The obtained results showed that phosphatidylcholine (16:0-18:1), phosphatidylethanolamine (18:0-20:4), phosphatidylinositol (18:0-18:2), phosphatidylserine (18:0-18:2), sphingomyelin (d18:1/16:0) and lysophosphatidylcholine (16:0) were the predominant species among the different classes of egg yolk phospholipids.