In vitro Antioxidant, Cytotoxic and Antidiabetic Activities of Protein Hydrolysates Prepared from Chinese Pond Turtle (Chinemys reevesii).

RESEARCH BACKGROUND: Cardiovascular diseases and diabetes are the biggest causes of death globally. Bioactive peptides derived from many food proteins using enzymatic proteolysis and food processing have a positive impact on the prevention of these diseases. The bioactivity of Chinese pond turtle muscle proteins and their enzymatic hydrolysates has not received much attention, thus this study aims to investigate their antioxidant, antidiabetic and cytotoxic activities. EXPERIMENTAL APPROACH: Chinese pond turtle muscles were hydrolysed using four proteolytic enzymes (Alcalase, Flavourzyme, trypsin and bromelain) and the degrees of hydrolysis were measured. High-performance liquid chromatography (HPLC) was conducted to explore the amino acid profiles and molecular mass distribution of the hydrolysates. The antioxidant activities were evaluated using various in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), hydroxyl radical scavenging activity, reducing capacity, chelating Fe2+ and lipid peroxide inhibition activity. Antidiabetic activity was evaluated using α-amylase inhibition and α-glucosidase inhibition assays. Besides, cytotoxic effect of hydrolysates on human colon cancer (HT-29) cells was assessed. RESULTS AND CONCLUSIONS: The amino acid composition of the hydrolysates revealed higher mass fractions of glutamic, aspartic, lysine, hydroxyproline and hydrophobic amino acids. Significantly highest inhibition of lipid peroxidation was achieved when hydrolysate obtained with Alcalase was used. Protein hydrolysate produced with Flavourzyme had the highest radical scavenging activity measured by DPPH (68.32%), ABTS (74.12%) and FRAP (A 700 nm=0.300) assays, α-glucosidase (61.80%) inhibition and cytotoxic effect (82.26%) on HT-29 cell line at 550 µg/mL. Hydrolysates obtained with trypsin and bromelain had significantly highest (p<0.05) hydroxyl radical scavenging (92.70%) and Fe2+ metal chelating (63.29%) activities, respectively. The highest α-amylase (76.89%) inhibition was recorded when using hydrolysates obtained with bromelain and Flavourzyme. NOVELTY AND SCIENTIFIC CONTRIBUTION: Enzymatic hydrolysates of Chinese pond turtle muscle protein had high antioxidant, cytotoxic and antidiabetic activities. The findings of this study indicated that the bioactive hydrolysates or peptides from Chinese pond turtle muscle protein can be potential ingredients in pharmaceuticals and functional food formulations.

Physicochemical properties, microstructure, and storage stability of Pulicaria jaubertii extract microencapsulated with different protein biopolymers and gum arabic as wall materials.

This study aimed to evaluate the possibility of using gum arabic (GA) with different protein materials namely whey protein isolate (WP), sodium caseinate (SC), and soybean protein (SP) as wall materials to encapsulate Pulicaria jaubertii extract (PJ) using freeze-drying. Four formulations of microencapsulation of Pulicaria jaubertii extract (MPJE) were produced, including WPGA-MPJE, SCGA-MPJE, SPGA-MPJE, and GA-MPJE. The formulations were stored at 4 °C and 25 °C for 28 days to assess the storage stability. The results indicated that mixtures of proteins with GA improved the physicochemical properties and bioactive content of the MPJE compared to GA-MPJE. The SCGA-MPJE formula showed optimal values of particle size (450.13 nm), polydispersity index (0.33), zeta potential (74.63 mV), encapsulation efficiency (91.07%), total phenolic content (25.51 g GAE g-1 capsules), and antioxidants compounds, as well as presented a lower release of bioactive composites with high oxidative stability during storage at 4 °C and 25 °C. The microstructure of MPJE formulations showed a flat surface without any visible cracking on surfaces. The microcapsules prepared from protein mixtures with GA, especially the SCGA-MPJE formula, are the most efficient in encapsulating the plant extract derived from the PJ, which could be useful for application in various industrial fields.

Stabilization of water-in-oil emulsion of Pulicaria jaubertii extract by ultrasonication: Fabrication, characterization, and storage stability.

This study investigated the effect of ultrasonic treatments on the properties and stability of the water-in-oil (W/O) emulsion of Pulicaria jaubertii (PJ) extract. The study used different ultrasound powers (0, 100, 200, 400, and 600 W) at two storage degrees (4 and 25 °C) for 28 days. The findings showed that the emulsifying properties were improved to different extents after ultrasonic treatments. The treatment at 600 W showed optimum particle size, polydispersity index, emulsifying property, viscosity properties, and release of total phenolic content than the other powers. However, the ultrasonic power of 400 W gave positive effects on creaming index and antioxidant release compared to 600 W. The emulsion stored at 4 °C presented higher stability than that stored at 25 °C during the 28 days of storage. Microscopically, the increase in sonication power up to 600 W reduced particle size and decreased flocculation, thus resulted in stable emulsions, which is desirable for its applications in food systems.

Supercritical fluid extraction of four aromatic herbs and assessment of the volatile compositions, bioactive compounds, antibacterial, and anti-biofilm activity.

Artemisia arborescens, Artemisia abyssinica, Pulicaria jaubertii, and Pulicaria petiolaris are fragrant herbs traditionally used in medication and as a food seasoning. To date, there are no studies on the use of supercritical fluids extraction with carbon dioxide (SFE-CO2) on these plants. This study evaluates and compares total phenolic content (TPC), antioxidant activity by DPPH• and ABTS•+, antibacterial, and anti-biofilm activities of SFE-CO2 extracts. Extraction was done by SFE-CO2 with 10% ethanol as a co-solvent. A. abyssinica extract had the highest extraction yield (8.9% ± 0.41). The GC/MS analysis of volatile compounds identified 307, 265, 213, and 201compounds in A. abyssinica, A. arborescens, P. jaubertii, and P. petiolaris, respectively. The P. jaubertii extract had the highest TPC (662.46 ± 50.93 mg gallic acid equivalent/g dry extract), antioxidant activity (58.98% ± 0.20), and antioxidant capacity (71.78 ± 1.84 mg Trolox equivalent/g dry extract). The A. abyssinica and P. jaubertii extracts had significantly higher antimicrobial activity and were more effective against Gram-positive bacteria. B. subtilis was the most sensitive bacterium. P. aeruginosa was the most resistant bacterium. P. jaubertii extract had the optimum MIC and MBC (0.4 mg/ml) against B. subtilis. All SFE-CO2 extracts were effective as an anti-biofilm formation for all tested bacteria at 1/2 MIC. Meanwhile, P. jaubertii and P. petiolaris extracts were effective anti-biofilm for most tested bacteria at 1/16 MIC. Overall, the results indicated that the SFE-CO2 extracts of these plants are good sources of TPC, antioxidants, and antibacterial, and they have promising applications in the industrial fields.

Effect of maltodextrin combination with gum arabic and whey protein isolate on the microencapsulation of gurum seed oil using a spray-drying method.

This study aimed to evaluate the potential of maltodextrin (MD) combination with gum arabic (GA), and whey protein isolate (WPI) on the microencapsulation of gurum seeds oil by a spray-drying method. Three formulations of protein-based (PB) (WPI: MD, 2:1), carbohydrate-based (CHOB) (GA: MD, 2:1), and mixed (MIX) (WPI: GA: MD, 1:1:1) wall materials were designed. The moisture content and water activity were in the range of 1.65-3.67% and 0.17-0.31, respectively, which is suitable for long-term storage. The best results were achieved when gurum seed oil was microencapsulated with carbohydrate-based, where it had the highest microencapsulation yield (92.80%) and microencapsulation efficiency (97.38%). Carbohydrate-based showed the highest relative crystallinity (32.25%) and the temperature of the glass transition (58.20 °C). FT-IR revealed that the oil was well encapsulated in the microcapsules. SEM of microcapsules showed spherical shapes without any apparent cracking on the surfaces. During the oxidative stability study, carbohydrate-based microencapsulation was the wall material that best protected the active materials against lipid oxidation.

Role of peppermint oil in improving the oxidative stability and antioxidant capacity of borage seed oil-loaded nanoemulsions fabricated by modified starch.

Borage seed oil (BSO) is one of the richest sources of γ-linolenic acid and linoleic acid, which are considered to retain plenty of health promoting benefits. However, its application in functional foods and dietary supplements remains limited owing to its superior vulnerability to oxidation. To solve this problem, ultrasound-assisted BSO-loaded nanoemulsions were prepared with modified starch incorporating different concentrations of peppermint oil (PO), as a natural antioxidant. The influence of different PO levels on the mean droplet size, rheology attributes, and oxidative stability of nanoemulsions stored at various temperatures (4, 25, and 40 °C) during 30 days storage was analyzed. In addition, DPPH and ABTS assays were used to determine the antioxidant activity and antioxidant capacity of BSO-loaded nanoemulsions, respectively. The optimized formulation (NE3; 5:5% v/v PO: BSO) exhibited a slight change in droplet size and oxidative stability at all temperatures during storage compared to other formulations. At a concentration of 328.08 µL/mL, formulation NE3 presented the minimum DPPH IC50 at 40 °C, which was lower than other formulations. The findings of this study revealed that the maximum retained antioxidant capacity (99.42 µg Trolox/mL) was related to NE3 comprising (5:5% v/v PO: BSO) stored at 40 °C for 30 days; which could be accredited to the role of PO as a natural antioxidant in order to improve the oxidative stability of nanoemulsion delivery system. Taken together, co-encapsulation of BSO and PO within nanoemulsions provides novel insights regarding the development of functional foods, dietary supplements and beverages.

Synergistic effects of amorphous OSA-modified starch, unsaturated lipid-carrier, and sonocavitation treatment in fabricating of Lavandula angustifolia essential oil nanoparticles.

This investigation aims to evaluate the synergistic effects of amorphous OSA-modified starch, unsaturated lipid-carrier (RBD-SFO), and high-energy microfluidization in synergy with the ultrasonic techniques in fabricating of Lavandula angustifolia essential oil (LAF-EO) nanoparticle. GC-MS and SEM techniques were employed to investigate the LAF-EO isolation method used. DLS analysis was employed along with CLSM and TEM techniques to investigate the physicochemical properties of nanoemulsion formulation (NE) matrices. The NE achieved the optimal spherical and size distributions of droplets (125.7 nm), Poly Dispersity Index (PdI) (0.183), and ζ-potential (-40.3 mV) when the contents of the formulation matrix were as follows: OSA-MS (2%), LAF-EO (1%), RBD-SFO (1%), and Tween-80 (1%). The findings of this work provide a new concept about the synergistic effects of amorphous OSA-modified starch and unsaturated lipid carrier as safe-grade macromolecules in the fabricating of LAF-EO nanoparticles. Besides, the application of the ultrasound cavitation phenomenon has been shown to have effective effect in reducing the droplet hydrodynamic diameter along with enhancing the distribution (PdI) and electrokinetic potential of the LAF-EO nanoparticles.

Microencapsulation of fingered citron extract with gum arabic, modified starch, whey protein, and maltodextrin using spray drying.

The aim of this study was to investigate the characteristics of the Fingered citron extract (FCE) microcapsules powders with different formulations of the encapsulation carrier agents [gum arabic (GA), maltodextrin (MD), modified starch (MS), and whey protein (WP)], which are obtained through spray drying. Encapsulation yield, encapsulation efficiency, moisture content, hygroscopicity, water activity, density properties, Carr's index, cohesiveness, flowability, porosity, wettability, solubility, color, particle size, thermal behavior, relative crystallinity, and micrographs were analyzed. Moreover, chemical structure of the microcapsule powder was identified using Fourier transform infrared spectroscopy (FT-IR). Best results were achieved when Fingered citron extract was encapsulated using gum arabic/maltodextrin/modified starch (GMS) formulation as carrier agents, where it had the highest encapsulation yield (89.39%) and encapsulation efficiency (87.20%). Furthermore, it achieved the best result in density properties, flowability, porosity, wettability, and relative crystallinity. These results have shown that using gum arabic/maltodextrin/modified starch formulation was superior for production of the Fingered citron extract microcapsules powders.