Effects Of Whey Protein Hydrolysate Research Articles

Antihypertensive effects of whey protein hydrolysate involve reshaping the gut microbiome in spontaneously hypertension rats

Novel angiotensin-converting enzyme (ACE) inhibitory peptides were identified from whey protein hydrolysates (WPH) in vitro in our previous study and the antihypertensive abilities of WPH in vivo were further investigated in the current study. Results indicated that WPH significantly inhibited the development of high blood pressure and tissue injuries caused by hypertension. WPH inhibited ACE activity (20.81 %, P < 0.01), and reduced renin concentration (P < 0.05), thereby reducing systolic blood pressure (SBP) (12.63 %, P < 0.05) in spontaneously hypertensive rats. The increased Akkermansia, Bacteroides, and Lactobacillus abundance promoted high short chain fatty acid content in feces after WPH intervention. These changes jointly contributed to low blood pressure. The heart weight and cardiomyocyte injuries (hypertrophy and degeneration) were alleviated by WPH. The proteomic results revealed that 19 protein expressions in the heart mainly associated with the wingless/integrated (Wnt) signaling pathway and Apelin signaling pathway were altered after WPH supplementation. Notably, WPH alleviated serum oxidative stress, indicated by the decreased malondialdehyde content (P < 0.01), enhanced total antioxidant capacity (P < 0.01) and superoxide dismutase activity (P < 0.01). The current study suggests that WPH exhibit promising antihypertensive abilities in vivo and could be a potential alternative for antihypertensive dietary supplements.

Whey protein hydrolysate maintains the homeostasis of muscle metabolism in exercise mice and releases potential anti-fatigue peptides after gastrointestinal digestion

Whey protein hydrolysate (WPH) has been proven to possess anti-fatigue effects, but its regulatory mechanism on muscle metabolism in exercise mice is unclear. In the present study, the effects of WPH on muscle metabolites in mice before and after exercise were explored using metabolomics techniques. Results showed that 34 muscle metabolites were significantly altered by exercise in the WPH group, which was less than 71 in the WP group and 101 in the control group. Specifically, supplementation of WPH significantly increased the levels of amino acid and carbohydrate metabolites (i.e., 1-methylhistamine, guanidoacetic acid, melibiose, and xylulose 5-phosphate) pre-exercise, thereby delaying energy depletion and maintaining metabolic homeostasis during exercise. Furthermore, simulated gastrointestinal digestion (SGID), peptidomics and bioinformatics tools were employed to screen potential peptides in WPH. A total of 37 peptides with 2–5 residues in WPH were resistant to SGID (peak intensity > 105). Among them, YGLF showed favorable potentials in physicochemical property, bioavailability, bioactivity, SwissTargetPrediction, and molecular docking. In addition, the anti-fatigue activity of YGLF was evaluated in vivo. YGLF (100 mg/kg) significantly increased the contents of glucose, glycogens, and amino acids and decreased the levels of lactate and urea nitrogen in mice after exercise. These findings may serve as a reference for the screening of anti-fatigue peptides.

Lactobacillus brevis M2-Fermented Whey Protein Hydrolysate Increases Slow-Wave Sleep via GABAA Receptors in Rodent Models.

In this study, we investigated the effects of whey protein hydrolysate (WPH) fermented with Lactobacillus brevis on sleep behavior and GABAergic mechanisms in rodent models. Fermentation converted the glutamate in WPH to high (3.15 ± 0.21 mg/mL) levels of γ-aminobutyric acid (GABA). Fermented WPH (WP-SF) enhanced sleep duration in mice by increasing GABA content in the brain. The increase in sleep duration induced by WP-SF resulted from an increase in delta wave activity during non-rapid eye movement sleep, and its sleep-promoting effect in a caffeine-induced insomnia model was characterized by an increase in delta waves. WP-SF increased GABAergic receptors at both mRNA and protein levels. Cotreatment with GABAA receptor antagonists abolished the sleep-promoting effects of WP-SF, indicating that WP-SF shares binding sites with antagonists on GABAA receptors. Collectively, WP-SF effectively increased sleep duration by enhancing delta wave activity through GABAergic activation; thus, it is suggested as a functional food-grade ingredient for promoting sleep.

Evaluation of the Improvement Effect of Whey Protein Poly-Peptides on Quality Characteristics of Repeated Freeze–Thawed Spanish Mackerel Surimi Balls

This investigation aimed to assess the effects of whey protein hydrolysate (WPH) on the oxidative stability of protein and the ability of Scomberomorus niphoniu surimi balls to retain water after repeated freeze-thaw (F-T) cycles. Ten percent natural whey peptides (NWP), 5% WPH, 10% WPH, 15% WPH, 0.02% butyl hydroxyl anisole (BHA), and a control group that did not receive any treatment were the six groups that were employed in the experiment. The cooking loss, water retention, total sulfhydryl content, and carbonyl content of each group were all measured. Notably, it was found that the surimi balls’ capacity to hold onto water and fend off oxidation was enhanced in a dose-dependent manner by the addition of WPH. Furthermore, the results showed that the 15% WPH added to the surimi balls effectively decreased protein oxidation in the F–T cycles and ameliorated the texture deterioration of surimi balls induced by repeated F–T, laying a theoretical foundation for the industrial application of WPH in surimi products.

Application of whey protein hydrolysate as a technological additive in the sour cream production

In the context of economic sanctions, the problem of supplying food additives, including stabilizers and thickeners, which are important in the production of fermented milk products, remains relevant for many enterprises; the development of a fermented milk product – sour cream, using whey protein hydrolysate as a technological additive is relevant. The purpose of the research was to study the effect of whey protein hydrolysate on the technological process of manufacturing a fermented milk product, sour cream, and the quality indicators of the final product. The results of the acidity study showed that in the experimental samples, the ripening process occurred faster than in the control sample. Based on the results of the research, it can be concluded that whey protein hydrolysate can be used as a technological additive in the production of fermented milk products to improve quality indicators.

Reducing Effects of Whey Protein Hydrolysate on Coloration of Cured Sausages.

Curing produces a characteristic pink color during meat processing through the production of nitrosyl myoglobin (NOMb), which requires nitric oxide (NO). Nitrites and nitrates in coloring agents are crucial NO sources; however, a reducing agent is necessary to facilitate their chemical conversion to NO. This study aimed to investigate the effect of the reducing properties of whey protein hydrolysate (WPH) on the reddening of cured meat products. Cured and cooked sausage models were treated with WPH, which enhanced the reddening of the meat color and increased the a* value in the models compared with that of the controls. Additionally, ethanol-extracted WPH induced Fe3⁺ reduction, lowered oxidation-reduction potential, and decreased nitrite (NO2-) levels. Moreover, ethanol-extracted WPH promoted the formation of NOMb in myoglobin solution. This effect was also observed when ethanol-extracted WPH treated with maleimide was used, implying that certain peptides rather than the thiol group of WPH are involved in promoting NOMb formation. Furthermore, the peptides that decreased NO2- levels were isolated from ethanol-extracted WPH, identified, and synthesized. These synthesized peptides, particularly the FFVAPFPEVFGK peptide, showed NO2--reducing activity. Hence, WPH may promote the coloration of cured meat products through the reducing potential of the peptides contained within.

Neuroprotective effect of whey protein hydrolysate containing leucine-aspartate-isoleucine-glutamine-lysine on HT22 cells in hydrogen peroxide–induced oxidative stress

This study aimed to investigate the neuroprotective effects of whey protein hydrolysate (WPH) containing the pentapeptide leucine-aspartate-isoleucine-glutamine-lysine (LDIQK). Whey protein hydrolysate (50, 100, and 200 µg/mL) demonstrated the ability to restore the viability of HT22 cells subjected to 300 µM hydrogen peroxide (H2O2)-induced oxidative stress. Furthermore, at a concentration of 200 µg/mL, it significantly reduced the increase in reactive oxygen species production and calcium ion (Ca2+) influx induced by H2O2 by 46.1% and 46.2%, respectively. Similarly, the hydrolysate significantly decreased the levels of p-tau, a hallmark of tauopathy, and BCL2 associated X (BAX), a proapoptosis factor, while increasing the protein levels of choline acetyltransferase (ChAT), an enzyme involved in acetylcholine synthesis, brain-derived neurotrophic factor (BDNF), a nerve growth factor, and B-cell lymphoma 2 (BCL2, an antiapoptotic factor. Furthermore, it increased nuclear factor erythroid 2-related factor 2 (Nrf2)-hemoxygenase-1(HO-1) signaling, which is associated with the antioxidant response, while reducing the activation of mitogen-activated protein kinase (MAPK) signaling pathway components, namely phosphor-extracellular signal-regulated kinases (p-ERK), phosphor-c-Jun N-terminal kinases (p-JNK), and p-p38. Column chromatography and tandem mass spectrometry analysis identified LDIQK as a compound with neuroprotective effects in WPH; it inhibited Ca2+ influx and regulated the BAX/BCL2 ratio. Collectively, WPH containing LDIQK demonstrated neuroprotective effects against H2O2-induced neuronal cell damage, suggesting that WPH or its active peptide, LDIQK, may serve as a potential edible agent for improving cognitive dysfunction.

Protective Effects of Whey Protein Hydrolysate, Treadmill Exercise, and Their Combination against Scopolamine-Induced Cognitive Deficit in Mice.

In this study, the potential of whey protein hydrolysate (WPH) and treadmill exercise to prevent cognitive decline was investigated, along with their neuroprotective mechanisms. Cognitive dysfunction was induced in mice with 1 mg/kg of scopolamine, followed by the administration of WPH at 100 and 200 mg/kg and/or treadmill exercise at 15 m/min for 30 min five days per week. Both WPH administration and treadmill exercise significantly improved the memory of mice with scopolamine-induced cognitive impairment, which was attributed to several key mechanisms, including a reduction in oxidative stress based on decreased levels of reactive oxygen species and malondialdehyde in the brain tissue and an increase in acetylcholine by increasing choline acyltransferase and decreasing acetylcholine esterase levels. Exercise and WPH also exerted neuroprotective effects by inhibiting the hyperphosphorylation of tau proteins, enhancing the expression of the brain-derived neurotrophic factor, and inhibiting apoptosis by reducing the Bax/Bcl2 ratio in conjunction with the downregulation of the mitogen-activated protein kinase pathway. Moreover, the impact of WPH and treadmill exercise extended to the gut microbiome, suggesting a potential link with cognitive improvement. These findings suggest that both WPH intake and treadmill exercise are effective strategies for mitigating cognitive impairment, providing promising avenues for treating neurodegenerative diseases.

Whey Protein Hydrolysate Improved the Structure and Function of Myofibrillar Protein in Ground Pork during Repeated Freeze-Thaw Cycles.

Whey protein hydrolysate (WPH) has made a breakthrough in inhibiting oxidative deterioration and improving the quality of meat products during storage. Based on our previous study of extracting the most antioxidant active fraction I (FI, the molecular weight < 1 kDa) from whey protein hydrolysates of different molecular weights, the present study continued to delve into the effects of WPH with fraction I on the structure and function of myofibrillar proteins (MP) in ground pork during the freeze-thaw (F-T) cycles. With the number of F-T cycles raised, the total sulfhydryl content, the relative contents of α-helix, Ca2+-ATPase activity, K+-ATPase activity, solubility, emulsion activity index (EAI), and emulsion stability index (ESI) of MP gradually decreased. Conversely, the carbonyl content and the relative content of random curl showed an increasing trend. In particular, the damage to the structure and the function of MP became more pronounced after three F-T cycles. But, during F-T cycles, FI stabilized the structure of MP. Compared to the control group, the 10% FI group showed a remarkable improvement (p < 0.05) in the total sulfhydryl content, Ca2+-ATPase activity, K+-ATPase activity, solubility, EAI and ESI after multiple F-T cycles, suggesting that 10% FI could effectively inhibit protein oxidation and had the influence of preserving MP function properties. In conclusion, WPH with fraction I can be used as a potential natural antioxidant peptide for maintaining the quality of frozen processed meat products.

Preparation of β-lactoglobulin-derived tryptophan peptide and its effect on anxiety-like behaviors in Zebrafish.

This study aimed to obtain three Trp-containing peptides from β-lactoglobulin and study their effects on anxiety-like behaviors in zebrafish. Three Trp-containing peptides were prepared from β-lactoglobulin by selective enzymatic hydrolysis and identified by UPLC-Q-TOF MS/MS. The anxiety-like behaviors of zebrafish were reduced after two weeks of administrated of β-lactoglobulin Trp peptides (LAWP), VAGTWY, VAGTW and G TW(concentration of 56 μg/mL or 500 μg/mL). As an index of serotonergic activity, we assessed the enhancing abilities of 5-HT synthesis. The treatment remarkably enhanced the 5-HT synthesis by upregulation of Trp concentration and Trp hydroxylase activation. In addition, this study further validated the anti-anxiety effects of whey protein hydrolysate with a high Trp index in animal and the experimental results were consistent with those reported in previous studies. Our results showed that β-lactoglobulin Trp peptides ingestion has a significant anti-anxiety effect as evidenced by the increasing Trp concentration, TPH activation and 5-HT level compared to the control group, with the VAGTW being the more effective.

Protective effects of whey protein hydrolysate on Bifidobacterium animalis ssp. lactis Probio-M8 during freeze-drying and storage.

We evaluated the potential of whey protein hydrolysate as a lyoprotectant for maintaining the cell viability of Bifidobacterium animalis ssp. lactis Probio-M8 during freeze-drying and subsequent storage. The moisture content and water activity of the lyophilized samples treated by different concentrations of whey protein hydrolysate were ≤5.23 ± 0.33 g/100 g and ≤0.102 ± 0.003, respectively. During storage at 25°C and 30°C, whey protein hydrolysate had a stronger protective effect on B. lactis Probio-M8 than the same concentration of whey protein. Using the Excel tool GinaFit, we estimated the microbial inactivation kinetics during storage. Whey protein hydrolysate reduced cell damage caused by an increase in temperature. Whey protein hydrolysate could protect cells by increasing the osmotic pressure as a compatible solute. Whey protein hydrolysate improved cell membrane integrity and reduced the amounts of reactive oxygen species and malondialdehyde produced. The findings indicated that whey protein hydrolysate was a novel antioxidant lyoprotectant that could protect probiotics during freeze-drying and storage.

In Vitro Gut Fermentation of Whey Protein Hydrolysate: An Evaluation of Its Potential Modulation on Infant Gut Microbiome.

Whey protein and its hydrolysate are ubiquitously consumed as nutritional supplements. This study aimed to evaluate the potential effect of whey protein hydrolysate (WPH) on the infant gut microbiome, which is more variable than that of adults. Colonic fermentation was simulated through a static digestion model and fecal culture fermentation, using feces from normal infants aged from 1–3 years old. During in vitro gut fermentation, measurements of short-chain fatty acids (SCFA) concentrations and 16S rRNA amplicon sequencing were performed. Additionally, the growth curves of cultivated probiotics were analyzed to evaluate the prebiotic potential of WPH. Besides the decline of pH in fermentation, the addition of WPH induced a significant increase in the SCFA production and also the relative abundance of Proteobacteria, Bacteroides, and Streptococcus (p < 0.05). The lower ratio of Firmicutes/Bacteroidetes in WPH-supplemented samples indicated the positive modulation of WPH on the gut microbiota, which could benefit the energy balance and metabolism of infants. The stimulation effect of WPH on the probiotics (particularly Lactobacillus acidophilus NCFM) during cultivation implied the prebiotic potential as well. Our findings shed light on WPH as a valuable dietary supplement with not only enriched resources of essential amino acids but also the potential to restore the infant gut microbiome.

Anti-hyperuricemic and nephroprotective effects of whey protein hydrolysate in potassium oxonate induced hyperuricemic rats.

Hyperuricemia (HUA) is a serious public health concern globally that needs to be solved. It is closely related to gout and other metabolic diseases. To develop a safe and effective dietary supplementation for alleviating HUA, we investigated the effects of whey protein hydrolysate (WPH) on HUA and associated renal dysfunction and explored their underlying mechanism. Potassium oxonate was used to induce HUA in model rats, who were then administered WPH for 21 days. The results showed that WPH significantly inhibited xanthine oxidase and adenosine deaminase activity in serum and liver, decreased uric acid (UA), creatinine, and blood urea nitrogen levels in serum, and increased the UA excretion in urine. In addition, WPH downregulated the expression of urate transporter 1 and upregulated the expression of organic anion transporter 1, adenosine triphosphate binding cassette subfamily G member 2, organic cation/carnitine transporters 1 and 2, and organic cation transporter 1 in kidneys. These findings demonstrated for the first time that WPH could alleviate HUA by inhibiting UA production and promoting UA excretion, and improve the renal dysfunction caused by HUA. Thus, WPH may be a potential functional ingredient for the prevention and treatment of HUA and associated renal dysfunction. © 2021 Society of Chemical Industry.

Stimulating effect of whey protein hydrolysate on bone growth in MC3T3-E1 cells and a rat model

This study was conducted to investigate the effect of whey protein hydrolysate (WPH) on osteogenic cell differentiation and its growth-promoting effects in rats. Alkaline phosphatase (ALP) activity and calcium deposition were measured by treating MC3T3-E1 cells with WPH, and mRNA and protein levels of factors related to osteoblast differentiation were assessed. ALP activity and calcium deposition were significantly increased in the WPH group (p < 0.001). These findings were confirmed by the upregulation of ALP, bone morphogenic protein, bone sialoprotein, and collagen at the mRNA and protein levels. Furthermore, to confirm the growth-promoting effect of WPH, bone growth was analyzed by administering 3-week-old Sprague-Dawley rats with whey protein or WPH. Moreover, serum levels of calcium, ALP, and insulin-like growth factor-1 (IGF-1) were analyzed, bone analysis was performed using micro-CT, and the size of the growth plate was measured by Cresyl violet staining. When rats were administered with a high dose of WPH (600 mg per kg per day), calcium levels decreased significantly, while ALP levels (1.14-fold; p < 0.01), IGF-1 levels, tibia length, and growth plate height increased significantly compared to those in the control group. Collectively, WPH has shown to be effective in bone differentiation and bone growth.

Peynir Altı Suyu Proteini Hidrolizatının Ayranda Yoğurt Starter Kültürleri ve Probiyotik Bakteriler Üzerine Etkilerinin İncelenmesi

This study aims to monitor the effect of whey protein hydrolysate (WPH) on the growth and activity of probiotic bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii spp. bulgaricus, Lactobacillus acidophilus, and Bifidobacterium lactis) in ayran, and also to enhance the functionality of ayran by addition of the probiotics. Effects of WPH and probiotics addition as 0%, 0.25%, 0.5% and 1% on the growth of probiotic bacteria were studied before and after the fermentation and 7th, 14th, and 21st days of the storage. WPH and probiotics had a significant effect (p&amp;lt;0.05) on the growth of target microorganisms and the biochemical variables. The addition of WPH and probiotics significantly (p&amp;lt;0.05) increased the growth of probiotics compared to the control after the fermentation up to the 21st day of storage. The samples containing different WPH levels had lower (p&amp;lt;0.05) pH levels compared to the control during the study time. The mixing of WPH at 1% had the highest total solids and protein content compared to the control samples. The WPH decreased (p&amp;lt;0.05) the viscosity and Hunter color parameters in added the samples. Results indicated that WPH had great potential for enhancing the growth of probiotic bacteria and the nutritional of ayran.

Effects of whey protein hydrolysate on growth promotion and immunomodulation in mouse pups in artificial rearing system.

This study aimed to investigate the effects of whey protein hydrolysate (WPH) on the growth and immunity of mouse pups in artificial rearing (AR) system. Mouse pups were reared in the AR system with artificial milk including 5% WPH (AR with WPH) or not (AR without WPH), and the remaining pups were reared by their mother (dam) for 14days after birth. The body weight change and body weight gain rates in the AR with WPH group were significantly higher than those observed in the AR without WPH group and similar to those in the dam group. Moreover the feed and protein efficiencies in the AR with WPH group were significantly higher than those of the AR without WPH group. In addition, the supplement of WPH in the AR system was shown to significantly elevate the number of CD3+ CD8+ , B220+ CD19+ , IA/IE+ CD11c+ , and CD11b+ in the thymocyte and/or splenocyte, and the thymus weight. Furthermore, MALDI-TOF/MS analysis identified the amino acid sequences corresponding to some peptides, and indicated that VRTPEVDDE had the highest relative intensity among the peptides from tested WPH. Therefore, WPH would be required to not only promote growth, but also exert immunomodulatory activities in mouse pups in AR system.

Suppressive effects of whey protein hydrolysate on sucrose-induced hyperglycemia in silkworms.

Silkworms are useful for evaluating substances that suppress postprandial hyperglycemia by oral administration. In this study, orally administered whey protein hydrolysate (WPH), obtained by enzymatic treatment of whey protein, suppressed sucrose-induced hyperglycemia in silkworms in a dose-dependent manner. WPH also inhibited glucose-induced hyperglycemia in silkworms. These findings suggest that WPH contains a bioactive peptide that inhibits glucose uptake from the intestinal tract and thereby suppresses sucrose-induced hyperglycemia.

Effects of Whey Protein Hydrolysate Ingestion on Postprandial Aminoacidemia Compared with a Free Amino Acid Mixture in Young Men.

To stimulate muscle protein synthesis, it is important to increase the plasma levels of essential amino acids (EAA), especially leucine, by ingesting proteins. Protein hydrolysate ingestion can induce postprandial hyperaminoacidemia; however, it is unclear whether protein hydrolysate is associated with higher levels of aminoacidemia compared with a free amino acid mixture when both are ingested orally. We assessed the effects of whey protein hydrolysate (WPH) ingestion on postprandial aminoacidemia, especially plasma leucine levels, compared to ingestion of a free amino acid mixture. This study was an open-label, randomized, 4 × 4 Latin square design. After 12–15 h of fasting, 11 healthy young men ingested the WPH (3.3, 5.0, or 7.5 g of protein) or the EAA mixture (2.5 g). Blood samples were collected before ingestion and at time points from 10 to 120 min after ingestion, and amino acids, insulin, glucose and insulin-like growth factor-1 (IGF-1) concentrations in plasma were measured. Even though the EAA mixture and 5.0 g of the WPH contained similar amounts of EAA and leucine, the WPH was associated with significantly higher plasma EAA and leucine levels. These results suggest that the WPH can induce a higher level of aminoacidemia compared with a free amino acid mixture when both are ingested orally.

Viability and growth promotion of starter and probiotic bacteria in yogurt supplemented with whey protein hydrolysate during refrigerated storage.

The effect of whey protein hydrolysate (WPH) addition on growth of standard yoghurt cultures and Bifidobacterium adolescentis during co-fermentation and its viability during storage at 4ºC in yoghurts has been evaluated. WPH was obtained with the use of serine protease from Y. lipolytica yeast. Stirred probiotic yoghurts were prepared by using whole milk standardized to 16% of dry matter with the addition of either whey protein concentrate, skim milk powder (SMP), WPH-SMP (ratio 1:1), WPH. The hydrolysate increased the yoghurt culture counts at the initial stage of fermentation and significantly inhibited the decrease in population viability throughout the storage at 4ºC in comparison to the control. The post-fermentation acidification was also retarded by the addition of WPH. The hydrolysate did not increase the Bifidobacterium adolescentis counts at the initial stage. However, the WPH significantly improved its viability. After 21 days of storage, in the yogurts supplemented with WPH, the population of these bacteria oscillated around 3.04 log10 CFU/g, while in samples where SMP or whey protein concentrate was used, the bacteria were no longer detected.

Whey protein hydrolysates enhance water absorption in the perfused small intestine of anesthetized rats.

We evaluated the effect of whey protein hydrolysates (WPH) on the water absorption rate in the small intestine using a rat small intestine perfusion model. The rate was significantly higher with 5g/L WPH than with 5g/L soy protein hydrolysates or physiological saline (p < 0.05). WPH dose-dependently increased the water absorption rate in the range of 1.25-10.0g/L. WPH showed a significantly higher rate than an amino acid mixture whose composition was equal to that of WPH (p < 0.05). The addition of 4-aminomethylbenzoic acid, an inhibitor of PepT1, significantly suppressed WPH's enhancement of water absorption (p < 0.05). The rate of water absorption was significantly correlated with that of peptides/amino acids absorption in WPH (r = 0.82, p < 0.01). These data suggest that WPH have a high water absorption-promoting effect, to which PepT1 contributes.