This study aimed to discover and prepare novel angiotensin converting enzyme (ACE) inhibitory peptides from almond protein and further evaluate the effect on endothelial function of human umbilical vascular endothelial cells (HUVECs). Almond protein was hydrolyzed using a two-stage alcalase-protamex hydrolysis process, and the hydrolysates were subjected to a series of separations, ultrafiltration, gel filtration chromatography, and reversed-phased preparative chromatography, to obtain the active peptides. Seven ACE inhibitory fractions with the molecular weight below 1.5 kDa were isolated and prepared, and two purified ACE inhibitory peptides with the IC50 values of 67.52 ± 0.05 and 43.18 ± 0.07 μg mL(-1), were identified as Met-His-Thr-Asp-Asp and Gln-His-Thr-Asp-Asp, respectively. Then the effect of two ACE inhibitory peptides on the endothelial function of HUVECs was evaluated. Results showed that the two potent ACE inhibitory peptides significantly regulated the release of nitric oxide and endothelin in HUVECs. These results suggest that almond peptides have potential as an antihypertensive nutraceuticals or a functional food ingredient.
A novel method for protein extraction from sweet almonds with aqueous polyethylene glycol (PEG) as solvent and recovery from the extraction solution was developed. The extraction yields of different solvents, such as sodium hydroxide, sodium chloride, PEG 200, PEG 400, and PEG 600 aqueous solutions, were investigated and PEG 200 showed the highest extraction efficiency. The PEG-based microwave-assisted extraction (MAE) parameters were then optimized using response surface methodology. Under optimum condition, PEG 200 concentration of 25 % (w/w), liquid to solid ratio of 22 mL g −1 , microwave power of 120 W, extraction temperature of 45°C, and extraction time of 4 min, the average extraction yield was 93.75 ± 3.15 %. Subsequently, the almond protein was recovered from the extraction solution containing PEG with an isoelectric point-ethanol synergy precipitation protocol. The combined technique integrated the speed of isoelectric point precipitation with the completeness of alcohol precipitation. The recovery of almond protein was 98.81 % with a time of 3-5 min. The proposed PEG-based MAE and synergy precipitation protocol provide a rapid and effective method for almond protein extraction and recovery and have the potential to be used for other plant proteins.
Carbon-based nanomaterials have recently attracted tremendous attention in adsorption, separation and biological fields. However, such modification is not always straightforward when the surface is not chemically reactive. Given this reason, most carbon materials modification processes employ reactive linkers or coupling agents, which are complicated and time-consuming. Herein, we report on a dopamine-polyethyleneimine (PEI) coating strategy to fabricate micrometer-sized magnetic carbon fiber (MSMCF)-based extracorporeal blood-cleansing sorbent for hemoperfusion. Results showed that the dopamine/PEI-coated MSMCF had a twisted fiber shape with a size range of 80-120 μm in diameter and porous structure with a specific surface area of 146 m g. Adsorption behavior of dopamine/PEI-modified MSMCF was examined by using bilirubin as a toxin model compound. Equilibrium data were well fitted to the Langmuir isotherm model with a maximal adsorption capacity of 335.1 mg g at ambient temperature. The as-obtained material had relatively high bilirubin adsorption selectivity against albumin at a normal albumin concentration. In particular, the dopamine/PEI-coated MSMCF has excellent adsorption capacity and hemocompatibility compared to the MSMCF decorated only by dopamine or PEI. Therefore, this work may pave the way for enhancing the property of the extracorporeal blood-cleansing sorbent during hemoperfusion.
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