Hypertension is the major controllable risk factor associated with cardiovascular disease (CVD) events such as myocardial infarction, stroke, heart failure, and end-stage diabetes. A 5 mm Hg decrease in blood pressure has been equated with approximately 16% decrease in CVD. In the U.S. alone current annual antihypertensive drug costs are approximately dollars 15 billion. The renin-angiotensin-aldosterone system is a target for blood pressure control. Cleavage of angiotensinogen by renin produces angiotensin I which is subsequently hydrolyzed by angiotensin-I-converting enzyme (ACE) to angiotensin II (a potent vasoconstrictor). Various side effects are associated with the use of ACE inhibitory drugs in the control of blood pressure including hypotension, increased potassium levels, reduced renal function, cough, angioedema, skin rashes, and fetal abnormalities. Milk proteins, both caseins and whey proteins, are a rich source of ACE inhibitory peptides. Several studies in spontaneously hypertensive rats show that these casokinins and lactokinins can significantly reduce blood pressure. Furthermore, a limited number of human studies have associated milk protein-derived peptides with statistically significant hypotensive effects (i.e., lower systolic and diastolic pressures). The advent of effective milk protein based functional food ingredients/nutraceuticals for the prevention/control of blood pressure therefore has the potential to significantly reduce global healthcare cost.

Food proteins contain latent biofunctional peptide sequences within their primary structures which may have the ability to exert a physiological response in vivo. A large range of biofunctional peptides have been isolated from food proteins including opioid, immunomodulatory, antimicrobial, mineral binding, growth and muscle stimulating, anti-cancer, proteinase and angiotensin converting enzyme (ACE, EC 3.4.15.1) inhibitory peptides. The biofunctional peptide activity currently most studied in food proteins appears to be those that inhibit ACE. ACE plays a central role in the regulation of blood pressure (BP) through the production of the potent vasoconstrictor, angiotensin (Ang) II , and the degradation of the vasodilator, bradykinin (BK). ACE inhibitory peptides may therefore have the ability to lower BP in vivo by limiting the vasoconstrictory effects of Ang II and by potentiating the vasodilatory effects of BK. These ACE inhibitory peptides can be enzymatically released from intact proteins in vitro and in vivo during food processing and gastrointestinal digestion, respectively. ACE inhibitory peptides may be generated in or incorporated into functional foods in the development of 'natural' beneficial health products. Several products are currently on the market or are in development that contain peptide sequences which have ACE inhibitory properties. Detailed human studies are required in order to demonstrate the efficacy of these bioactive peptides prior to their widespread utilisation as physiologically beneficial functional foods/food ingredients.

8The ultrasonic effect on the physicochemical and emulsifying properties of three animal proteins, 9 bovine gelatin (BG), fish gelatin (FG) and egg white protein (EWP), and three vegetable proteins, pea protein 10 isolate (PPI), soy protein isolate (SPI) and rice protein isolate (RPI), was investigated. Protein solutions (0.1 -11 10 wt. %) were sonicated at an acoustic intensity of ~34 W cm -2 for 2 minutes. The structural and physical 12 properties of the proteins were probed in terms of changes in size, hydrodynamic volume and molecular 13 structure using DLS and SLS, intrinsic viscosity and SDS-PAGE, respectively. The emulsifying performance of 14 ultrasound treated animal and vegetable proteins were compared to their untreated counterparts and Brij 97. 15Ultrasound treatment reduced the size of all proteins, with the exception of RPI, and no reduction in the 16 primary structure molecular weight profile of proteins was observed in all cases. Emulsions prepared with all 17 untreated proteins yielded submicron droplets at concentrations ≤ 1 wt. %, whilst at concentrations > 5 wt. % 18 emulsions prepared with EWP, SPI and RPI yielded micron sized droplets (> 10 μm) due to pressure 19 denaturation of protein from homogenisation. Emulsions produced with sonicated FG, SPI and RPI had the 20 similar droplet sizes as untreated proteins at the same concentrations, whilst sonicated BG, EWP and PPI 21 emulsions at concentrations ≤ 1 wt. % had a smaller droplet size compared to emulsions prepared with their 22 untreated counterparts. This effect was consistent with the observed reduction in the interfacial tension between 23 these untreated and ultrasound treated proteins.

Milk proteins contain regions within their primary structures that encrypt for many latent biological activities. The beneficial health effects associated with some fermented dairy products may, in part, be attributed to the release of bioactive peptide sequences during the fermentation process. Peptides displaying opioid, mineral binding, cytomodulatory and hypotensive activities, for example, have been identified in cheese and yogurt. Much effort has to date concentrated on the release of angiotensin‐converting enzyme (ACE) inhibitory peptides due to their potential to act as hypotensive agents. Peptide fractions obtained by hydrophobic interaction chromatography of different cheese varieties (Blue, Camembert, Edam, Emmental, Gouda and Havarti) were reported to give systolic blood pressure (SBP) decreases in spontaneously hypertensive rats (SHR) ranging from 7.1 to 29.3 mm mercury (Hg). Skim milks fermented with various strains of Lactobacillus helveticus, and in one case also with Saccharomyces cerevisiae, have been reported to display SBP decreases in mildly hypertensive human volunteers ranging from 4.6 to 14.1 mmHg. These human hypotensive effects have, in part, been attributed to the release of potent casein‐derived tripeptide inhibitors of ACE during fermentation. In general, the likelihood of any bioactive peptide released during fermentation mediating a physiological response is dependent on the ability of that peptide to reach an appropriate target site. Therefore, peptides may need to be resistant to further degradation by gastrointestinal and serum proteinases/peptidases following oral ingestion in order to display a functional food effect.

The aggregation of amyloid-β (Aβ) peptides plays a crucial role in the etiology of Alzheimer's disease (AD). Recently, it has been reported that an A2T mutation in Aβ can protect against AD. Interestingly, a nonpolar A2V mutation also has been found to offer protection against AD in the heterozygous state, although it causes early-onset AD in homozygous carriers. Since the conformational landscape of the Aβ monomer is known to directly contribute to the early-stage aggregation mechanism, it is important to characterize the effects of the A2T and A2V mutations on Aβ₁₋₄₂ monomer structure. Here, we have performed extensive atomistic replica-exchange molecular dynamics simulations of the solvated wild-type (WT), A2V, and A2T Aβ₁₋₄₂ monomers. Our simulations reveal that although all three variants remain as collapsed coils in solution, there exist significant structural differences among them at shorter timescales. A2V exhibits an enhanced double-hairpin population in comparison to the WT, similar to those reported in toxic WT Aβ₁₋₄₂ oligomers. Such double-hairpin formation is caused by hydrophobic clustering between the N-terminus and the central and C-terminal hydrophobic patches. In contrast, the A2T mutation causes the N-terminus to engage in unusual electrostatic interactions with distant residues, such as K16 and E22, resulting in a unique population comprising only the C-terminal hairpin. These findings imply that a single A2X (where X = V or T) mutation in the primarily disordered N-terminus of the Aβ₁₋₄₂ monomer can dramatically alter the β-hairpin population and switch the equilibrium toward alternative structures. The atomistically detailed, comparative view of the structural landscapes of A2V and A2T variant monomers obtained in this study can enhance our understanding of the mechanistic differences in their early-stage aggregation.

The objectives of this study were to investigate the generation of beta-lactoglobulin fragment (142-148) (beta-LG f(142-148) during the hydrolysis of whey proteins, and the in vitro stability of this fragment upon incubation with gastrointestinal and serum proteinases and peptidases. An enzyme immunoassay (EIA) protocol was developed for the quantification of beta-LG f(142-148) in whey protein hydrolysates and in human blood serum. The minimum detection limit was 3 ng/mL. The level of the peptide in whey protein hydrolysates was influenced by the degree of hydrolysis (DH). As expected, highest levels of this peptide were found in hydrolysates generated with trypsin. Sequential incubation of hydrolysates at different DH values with pepsin and Corolase PP, to simulate gastrointestinal digestion, generally resulted in the degradation of beta-LG f(142-148) as determined by EIA. Reversed-phase HPLC and angiotensin-I-converting enzyme (ACE) activity assays demonstrated that synthetic beta-LG f(142-148) was rapidly degraded upon incubation with human serum. Furthermore, beta-LG f(142-148) could not be detected by EIA in the sera of 2 human volunteers following its oral ingestion or in sera from these volunteers subsequently spiked with beta-LG f(142-148). These in vitro results indicate that beta-LG f(142-148) is probably not sufficiently stable to gastrointestinal and serum proteinases and peptidases to act as an hypotensive agent in humans following oral ingestion. The in vitro methodology described herein has general application in evaluating the hypotensive potential of food protein-derived ACE inhibitory peptides.

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High-protein milk protein concentrate (MPC) and milk protein isolate (MPI) powders may have lower solubility than low-protein MPC powders, but information is limited on MPC solubility. Our objectives in this study were to (1) characterize the solubility of commercially available powder types with differing protein contents such as MPC40, MPC80, and MPI obtained from various manufacturers (sources), and (2) determine if such differences could be associated with differences in mineral, protein composition, and conformational changes of the powders. To examine possible predictors of solubility as measured by percent suspension stability (%SS), mineral analysis, Fourier transform infrared (FTIR) spectroscopy, and quantitative protein analysis by HPLC was performed. After accounting for overall differences between powder types, %SS was found to be strongly associated with the calcium, magnesium, phosphorus, and sodium content of the powders. The FTIR score plots were in agreement with %SS results. A principal component analysis of FTIR spectra clustered the highly soluble MPC40 separately from the rest of samples. Furthermore, 2 highly soluble MPI samples were clustered separately from the rest of the MPC80 and MPI samples. We found that the 900 to 1,200 cm⁻¹ region exhibited the highest discriminating power, with dominant bands at 1,173 and 968 cm⁻¹, associated with phosphate vibrations. The 2 highly soluble MPI powders were observed to have lower κ-casein and α-(S1)-casein contents and slightly higher whey protein contents than the other powders. The differences in the solubility of MPC and MPI were associated with a difference in mineral composition, which may be attributed to differences in processing conditions. Additional studies on the role of minerals composition on MPC80 solubility are warranted. Such a study would provide a greater understanding of factors associated with differences in solubility and can provide insight on methods to improve solubility of high-protein milk protein concentrates.