Preparation and characterisation of palm-based tocotrienol-carotenoid emulsion stabilised by hemp protein isolate-ginsenoside emulsifier

Hemp protein, with its important nutritional and industrial value, has trickled into the aisles of protein demand; however, its poor functional properties have largely limited its implementation in food. Herein, we aimed to modify hemp protein isolate (HPI) via glycosylation coupling with pullulan polysaccharide, and we subsequently characterized its structural and functional properties. The conjugation variables were HPI to pullulan ratio (i.e., 3:1, 2:1, 1:1, 1:2, and 1:3 w/w), incubation temperature (i.e., 50, 60, 70, 80, and 90 °C), and incubation time (i.e., 3, 6, 12, 24, and 48 h). Native HPI was used as a control for comparison purposes. We found that DG tended to decrease when the pullulan to HPI ratio was greater than 1:1 and when the temperature exceeded 80 °C. SDS-PAGE analysis shows that when the DG is increased, wider and heavier molecular weight bands emerge near the top of the running gel, while such observations were absent in the control. Further, glycosylation could loosen the HPI’s secondary and tertiary structures, as well as increase surface hydrophobicity. The solubility of HPI after glycosylation significantly increased (p < 0.05) at pH 7.0 compared to HPI without glycosylation. Emulsifying activity improved significantly (p < 0.05), with glycosylation with HPI–pullulan at a ratio of 1:3 showing maximum emulsifying activity of 118.78 ± 4.48 m2/g (HPI alone: 32.38 ± 3.65 m2/g). Moreover, the HPI–pullulan glycosylation time of 24 h showed maximum foaming activity (23.04 ± 0.95%) compared to HPI alone (14.20 ± 1.23%). The foaming stability of HPI (79.61 ± 3.33%) increased to 97.78 ± 3.85% when HPI–pullulan was conjugated using a glycosylation temperature of 80 °C. Compared with the un-glycated HPI, HPI–pullulan also increased WHC (4.41 ± 0.73 versus 9.59 ± 0.36 g/g) and OHC (8.48 ± 0.51 versus 13.73 ± 0.59 g/g). Intriguingly, correlation analysis showed that protein functional characteristics were significantly and positively correlated with DG. Overall, our findings support the notion that pullulan conjugation provides further functional attributes to the HPI, thereby broadening its potential implementation in complicated food systems.

Manothermosonication, high-pressure homogenization, and their combinations with pH-shifting improve the techno-functionality and digestibility of hemp protein

Reverse micelles extraction of hemp protein isolate: Impact of defatting process on protein structure, functionality, and aromatic profile

Hemp protein isolate-polysaccharide complex coacervates and their application as emulsifiers in oil-in-water emulsions

Production, digestibility and allergenicity of hemp (Cannabis sativa L.) protein isolates

Enzymatic hydrolysis of hemp ( Cannabis sativa L.) protein isolate by various proteases and antioxidant properties of the resulting hydrolysates

The aim of this study was to investigate the influence of the nutrient composition, protein profile and physicochemical properties of two commercial hemp protein ingredients, i.e., a hemp protein isolate (HPI) and a hemp protein concentrate (HPC) obtained by wet extraction and oil pressing, respectively, on their processability under high moisture extrusion (HME) conditions. The hemp protein ingredients were processed both individually and in combination with commercial soy (SPC) and pea (PPC) protein concentrates.HPC displayed the highest concentration of cysteine (1.6 g/100 g protein) and free thiols (15 nmol/mg protein) among the ingredients investigated, while UPLC-MS/MS analysis highlighted a higher relative concentration of 11S globulin (edestin) in HPI than HPC.The hemp protein ingredients displayed low solubility in water across the pH range 3–8 (5–9%) and their hydration capacity was closer to that of wheat gluten than SPC and PPC.HME trials were performed using a bench scale twin-screw extruder. HPI was not processable regardless of the conditions tested, which could be explained by the high concentration of edestin macromolecular complexes which hindered the formation of the desired fibrous texture. Conversely, HPC provided extrudates with promising textural properties both when processed individually and in combination with SPC or PPC. The peak force required to cut the extrudates throughout their entire thickness, as determined using a texture analyzer equipped with a light knife blade probe guillotine, increased with an increase in the relative concentration of HPC for blends with either SPC or PPC, that is with an increase in the relative importance of covalent bonds in fibrous structure formation.The outcomes of this study should be confirmed at larger scale to assess the potential of the derived extrudates to be used for the development of meat and fish analogues.

Defatting dehulled hemp seeds is a crucial step prior to protein extraction. However, conventional methods rely on flammable solvents, posing significant health, safety, and environmental concerns. Additionally, hemp protein has poor extractability, challenging functionality, and flavor limitations, restricting its broader application in foods. Accordingly, a two-phase natural deep eutectic solvent (NADES)-assisted extraction was evaluated as a solvent-free alternative for co-extracting protein and oil from full-fat hemp flour. In comparison to the reference hemp protein isolate (R-HPI), produced from hexane-defatted flour following conventional alkaline extraction, NADES-extracted hemp protein isolate (N-HPI) had significantly higher protein extraction yield and purity. N-HPI exhibited enhanced surface charge, lower hydrophobicity, and thus higher solubility at an acidic pH compared to R-HPI. N-HPI had a higher abundance of edestin and lower levels of vicilin-like proteins, which contributed to superior gelation compared to R-HPI. N-HPI, compared to R-HPI, contained lower levels of lipid-derived off-flavor compounds, such as aldehydes, alcohols, and ketones. These findings highlighted, for the first time, the potential of a two-phase NADES-assisted extraction as a sustainable alternate and effective process for producing high-quality, functional hemp protein. The development of such a green process is an impetus for broadening the applications of hemp protein in food systems.

Substitution of synthetic emulsifiers by a hemp protein in canola oil-in-water emulsions resembling salad dressing: comparison of rheological properties and physical stability.

There is an increased awareness of the incorporation of hemp protein in a commercial product on account of its high nutritional value and neutral flavor. One of the factors that impact the functional properties of hemp protein is the isolation conditions. In recent years, reverse micelles (RMs) has emerged as a powerful technique for extracting protein and enzyme because of low cost, convenience, and potential for scaling up in the manufacturing process. For oil crops, it generally requires long defatting processing before protein extraction. Therefore, it would be interesting to investigate whether the defatting of hempseed flour would impact on physicochemical properties of hemp protein using the RM extraction method. Our results revealed that high protein recovery yield (69.37-70.21%) and protein purity (92.76-95.2%) of hemp protein isolate (HPI) could be obtained through the RM technique using either non-defatted hemp flour (ND-HF) or defatted hemp flour (D-HF). Concerning the defatting processing, both HPIs showed great similarity in protein composition, and functional properties including solubility, thermal stability, foaming, and emulsifying properties. Interestingly, HPI obtained from ND-HF had higher essential amino acid ratio when compared to D-HF. Beyond that, distinct gelling behaviors (least gelling concentration, gel strength, and color) have been observed. The HPI gel obtained from ND-HF displayed a higher least gelling concentration (4%) with yellowish color than that of HPI gel from D-HF (2%). In terms of aroma profile, a significantly high number of aromatic compounds with a greater abundancy were detected in HPI from ND-HF (33) as compared to HPI isolated from D-HF (26), which contributed to the different process conditions and lipid content. The research findings provided an eco-friendly protein isolation method with premium functional attributes, especially for oil crops, which pave the way for the future application of RMs in a wide range of protein extraction.

Ginsenoside Rg_3, an active component of traditional Chinese medicine(TCM), was used as the substitute for cholesterol as the membrane material to prepare the ginsenoside Rg_3-based liposomes loaded with dihydroartemisinin and paclitaxel. The effect of the prepared drug-loading liposomes on triple-negative breast cancer in vitro was evaluated. Liposomes were prepared with the thin film hydration method, and the preparation process was optimized by single factor experiments. The physicochemical properties(e.g., particle size, Zeta potential, and stability) of the liposomes were characterized. The release behaviors of drugs in different media(pH 5.0 and pH 7.4) were evaluated. The antitumor activities of the liposomes were determined by CCK-8 on MDA-MB-231 and 4T1 cells. The cell scratch test was carried out to evaluate the effect of the liposomes on the migration of MDA-MB-231 and 4T1 cells. Further, the targeting ability of liposomes and the mechanism of lysosome escape were investigated. Finally, H9c2 cells were used to evaluate the potential cardiotoxicity of the preparation. The liposomes prepared were spheroid, with uniform particle size distribution, the ave-rage particle size of(107.81±0.01) nm, and the Zeta potential of(2.78±0.66) mV. The encapsulation efficiency of dihydroartemisinin and paclitaxel was 57.76%±1.38% and 99.66%±0.07%, respectively, and the total drug loading was 4.46%±0.71%. The accumulated release of dihydroartemisinin and paclitaxel from the liposomes at pH 5.0 was better than that at pH 7.4, and the liposomes could be stored at low temperature for seven days with good stability. Twenty-four hours after administration, the inhibition rates of the ginsenoside Rg_3-based liposomes loaded with dihydroartemisinin(70 μmol·L~(-1)) and paclitaxel on MDA-MB-231 and 4T1 cells were higher than those of the positive control(adriamycin) and free drugs(P&lt;0.01). Compared with free drugs, liposomes inhibited the migration of MDA-MB-231 and 4T1 cells(P&lt;0.05). Liposomes demonstrated active targeting and lysosome escape. In particular, liposomes showed lower toxicity to H9c2 cells than free drugs(P&lt;0.05), which indicated that the preparation had the potential to reduce cardiotoxicity. The findings prove that ginsenoside Rg_3 characterized by the combination of drug and excipient is an ideal substitute for lipids in liposomes and promoted the development of innovative TCM drugs for treating cancer.

Antioxidant and ACE-inhibitory activities of hemp (Cannabis sativa L.) protein hydrolysates produced by the proteases AFP, HT, Pro-G, actinidin and zingibain

(1) Background: Hemp seeds are a source of plant-based protein, making them an appropriate supplement to a plant-based diet. The current work was focused on the preparation of the protein isolate from the hemp seeds with eco-friendly and cheap technology. Moreover, it evaluated the physicochemical and functional properties of hemp protein isolate for its potential application in food manufacturing. (2) Methods: The protein content of hemp seeds has been isolated through two main steps: (1) extraction of the protein content of an alkaline pH (10–12); (2) precipitation of the extracted protein on an acidic pH as an isoelectric point (pH = 4.5). (3) Results: The edastin protein is the most predominant protein in the protein profile with a molecular weight of 58.1 KDa beside albumin with a molecular weight of 31.5 KDa. The FTIR spectrum detected the absorption peaks of the amide I at 1750 and 1600 cm−1, which pointed to C=O stretching while N-H stretching at 1650–1580 cm−1. The peak at 3250 is found to be related to N-H stretching of the aliphatic primary amine (3400–3300 cm−1) and the N-H stretching for the secondary (II) amine appeared at 3350–3310 cm−1. The Hemp protein isolate (HPI) showed a high content of arginine (15.52 g/100 g), phenylalanine + tyrosine (9.63 g/100 g), methionine + cysteine (5.49 g/100 g), leucine + isoleucine (5.21 g/100 g), and valine (4.53 g/100 g). It contains a moderate level of threonine (3.29 g/100 g) and lysine (2.50 g/100 g) with tryptophan as the limiting amino acid (0.22 g/100 g). The HPI showed an appropriate water-and-oil holding capacity (4.5 ± 2.95 and 2.33 ± 1.88 mL/g, respectively). The foaming capacity of the HPI was increased with increasing the pH values to reach the maximum value at pH 11 (67.23 ± 3.20%). The highest emulsion ability index of the HPI was noted at pH 9 (91.3 ± 2.57 m2/g) with low stability (19.15 ± 2.03). (4) Conclusions: A strong positive correlation (r = 0.623) was shown between protein concentration and solubility. The current easy-to-use, cheap, and eco-friendly technology provides the industrial sector with a cheap protein isolate for manufacturing protein-rich diet and beverages. The HPI showed a good nutritional quality and functional properties that might be helpful in utilizing it in different food products such as beverages and bakery products.

Hemp protein (HP), a plant-based protein with high nutritional value, shows promising potential for application in the production of meat analogues (MA). However, the formation mechanism of MA fiber structure produced by different proportions of HP and soy protein isolate (SPI) and its impact on the quality of MA remains unclear. In this study, MA with different HP/SPI ratios was prepared by high moisture extrusion (HME), and its texture and structure were evaluated. The results revealed that an appropriate amount of HP improve the textural properties of MA, promote the formation of fiber structure, and significantly improve its oil holding capacity (OHC). Compared with no addition of HP, the degree of texturization of MA with 40% HP was increased by 21.1%, and the OHC with 60% HP was increased by 79.9%. Furthermore, the addition of 40% HP promoted the formation of hydrogen bonds and disulfide bonds in the proteins in the cooling and extrusion zones, thereby forming MA with better fiber structure. The secondary structure also showed that the addition of HP formed more ordered structures (α-helix and β-sheet) of protein in the cooling and extrusion zone, which played an important role in maintaining the fiber structure of MA.

Ultrasonic treatment of emulsion gels with different soy protein-hemp protein composite ratios: Changes in structural and physicochemical properties