Effect Of Casein Phosphopeptides Research Articles

Promoting the Calcium-Uptake Bioactivity of Casein Phosphopeptides in vitro and in vivo.

Casein phosphopeptides have been studied widely for their ability to chelate calcium. However, systematic studies on the effects of casein phosphopeptides (CPP) on calcium absorption in vitro and in vivo are scarce. The purities of two commercially available products, CPP1 and CPP2, are 18.37 and 25.12%, respectively. Here, the in vitro calcium binding capacity of CPP2 was 142.56 ± 7.39 mg/g, which was higher than that of CPP1 (107.15 ± 6.27 mg/g). The calcium transport results in a Caco-2 monolayer model indicated that, relative to controls, CPP1 and CPP2 increased calcium transport by 21.78 and 53.68%, respectively. Subsequent animal experiments showed that the CPP2-Ca-H group (1% Ca, 0.4% CPP2) had significant increases in the femur index, serum Ca2+ and serum osteocalcin levels, and femoral Ca content. The CPP2-Ca-H animal also had decreased serum alkaline phosphatase levels, parathyroid hormone content, and urinary pyridinoline content. Overall, our results demonstrated that CPP2 had stronger effects on promoting calcium uptake than CPP1.

Studies on Immunomodulatory Effect of Casein Phospho Peptide Isolated from Cultured Dairy Product

Bioactive peptides have been defined as specific protein fragments that have an impact on body functions or conditions and may ultimately influence health. Fermented milk is a dairy product which has abundance of bioactive peptides. In this study, Casein Phospho peptide (CPP) was isolated by enzymatic hydrolysis of fermented milk using trypsin. The molecular weight of the Casein Phospho peptide was 3.5 KDa. The anti-bacterial activity of Casein Phospho peptide was determined using four pathogens such as, Escherichia coli, Bacillus cereus, Staphylococcus aureus and Salmonella enterica. Casein phospho peptide formed a zone of inhibition against the pathogens. The bioactive peptides were characterized using Fourier Transform Infra-Red Spectroscopy (FT-IR), Casein Phospho peptide had aliphatic amine, acetyl amino I and acetyl amino II functional groups. The HPLC analysis of Casein Phospho peptide revealed that the major amino acid present was L- Glutamic acid and the amino acid present in lesser concentration was Leucine. Peripheral Blood Mononuclear Cells (PBMCs) were isolated from human blood and the cells were treated with Casein Phospho peptide to assess the immunomodulatory effect.
 Casein Phospho peptide was able to produce a higher concentration of IL-10 anti-inflammatory cytokines when treated with PBMCs.

Effect of Casein Phosphopeptides on In Vitro Solubilization of Calcium in Fortified Cheese Pizza

Cheese pizzas fortified with casein phosphopeptide (CPP) and calcium were subjected to an in vitro digestion to assess whether CPP could prevent the precipitation of calcium. The total calcium content of the cheese pizzas was adjusted to 1,000 mg per pizza (~370 g) with the addition of calcium originating from eggshells. Two levels of trypsin-digested caseins (367 and 459 mg), with a CPP content of ~20%, were added to each pizza. The in vitro digested pizzas were then centrifuged and the supernatant was mixed with Na2HPO3 at 37°C to estimate the possible soluble effect of CPP on calcium. After 24 h of reaction, the solution was centrifuged and the calcium content in the resultant supernatant was analyzed by inductively coupled plasma-atomic emission spectroscopy. One-way statistical analyses showed that CPP had a positive effect on the solubilization of calcium against phosphate (p<0.05). Cheese pizza supplemented with 459 mg of CPP powder was able to prevent precipitation of calcium by 98.8%, whereas no CPP-added cheese pizza solubilized 86.4% of the calcium. A sensory test was also carried out, revealing that panelists could not discern the bitter taste of the CPP added to the pizzas.

Effects of casein phosphopeptides on calcium absorption and metabolism bioactivity in vitro and in vivo.

Casein phosphopeptides (CPPs) are a series of peptides containing serine phosphate, which prevents calcium precipitation in the small intestine, so that it can be absorbed. Calcium metabolism studies have been typically carried out using traditional metabolic balance tests. Non-radioactive stable isotopes have rarely been used to examine the in vivo absorption and metabolism of calcium in animal models due to their high cost and the difficulty in their accurate detection. However, they have recently attracted intense research attention. In this study, we developed InertSep ME-1 using a chelating resin to effectively separate 42Ca from spectral-interference in the process of ICP-MS detection. The method effectively removes interfering ions like potassium and sodium and greatly improves the accurate detection of the calcium ion isotope. We also investigated the absorption, distribution, and metabolism of different CPPs through both in vitro cell experiments and in vivo animal experiments. The results indicate that CPPs strongly promote calcium absorption, especially the P5 active monomer component. The results of our in vivo studies show that the calcium isotope can be absorbed from the small intestine into the blood. Then, one part is transported to various organs through tissue fluids while another part is excreted into the urine through the kidneys. In general, our results reveal that CPPs promote the absorption of calcium significantly and positively affect calcium metabolism.

Effect of casein phosphopeptide - amorphous calcium phosphate containing chewing gum on salivary concentration of calcium and phosphorus: An in-vivo study

Caries clinical trials of sugar-free chewing gum have shown that the gum is noncariogenic and in fact has anticariogenic effect through the stimulation of saliva. Sugar-free gums, therefore, may be an excellent delivery vehicle for safe and effective additive, capable of promoting enamel remineralization. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) nanocomplexes incorporated into sugar-free chewing gum have shown to remineralize enamel subsurface lesions in situ. So this study was conducted to evaluate the effect of CPP-ACP containing sugar-free chewing gum on salivary concentration of calcium and phosphorous. Unstimulated saliva from each 24 selected subjects was collected. Then each subject was given two pellets of chewing gum containing CPP-ACP and asked to chew for a period of 20 min, after which saliva samples were collected from each individual. Once all the samples were collected they were assessed for calcium and phosphorous concentration using affiliated reagent kits and photometer. Data obtained were analyzed using student's paired t test. Significant difference was found in the calcium and phosphorus concentration of saliva before and after chewing CPP-ACP containing chewing gum. Chewing of CPP-ACP containing chewing gum showed a significant increase in the salivary concentration of calcium for a prolonged period of time hence it may help in the remineralization of tooth surfaces.

Effect of Casein phosphopeptide on Phagocytic Capacity and Intracellular Oxidative Function of Polymorphonuclear Neutrophils

Effect of Casein phosphopeptide on Phagocytic Capacity and Intracellular Oxidative Function of Polymorphonuclear Neutrophils

Effect of casein phosphopeptide - amorphous calcium phosphate on hardness of enamel eroded by a cola drink

Effect of casein phosphopeptide - amorphous calcium phosphate on hardness of enamel eroded by a cola drink

Effect of casein phosphopeptide and 25-hydroxycholecalciferol on tibial dyschondroplasia in growing broiler chickens

1. An experiment was conducted to investigate the influence of dietary casein phosphopeptides and 25-hydroxycholecalciferol on the incidence of tibial dyschondroplasia (TD) in 14-d-old commercial broiler chickens. 2. Three hundred and twenty broiler chicks (one day old) were randomly allocated to one of 4 dietary treatments. A standard broiler diet was used as the control with the three experimental treatments receiving the control diet supplemented with 10 g casein phosphopeptide/kg, 14 g casein phosphopeptide/kg or 69 µg 25-hydroxycholecalciferol/kg. 3. Those birds fed the diets supplemented with 14 g casein phosphopeptide/kg or 25-hydroxycholecalciferol had a lower incidence of TD than both the control and 10 g casein phosphopeptide/kg treatments when assessed grossly. 4. The body weight of birds fed the 10 g casein phosphopeptide/kg diet or the 25-hydroxycholecalciferol diet was higher than birds fed the control diet. Although not significant, the body weight of birds fed the 14 g casein phosphopeptide/kg diet was also greater than the control birds. 5. The current experiment demonstrated that both casein phosphopeptide and 25-hydroxycholecalciferol can reduce the incidence of TD in the young broiler chicken. More research is required to explain the unexpected increase in body weight described above.

Effect of caseinphosphopeptides on inhibition of calcium intestinal absorption due to phosphate

Caseinphosphopeptides (CPPs) are peptides resulting from enzyme digestion of whole casein, characterized by the presence of an aminoacid cluster, negatively charged at physiological intestinal pH, that binds cations. The aim of the present study was to compare Ca absorption through the everted small intestine of rats from CaCl 2 or Ca-CPPs solutions, in absence of inorganic phosphate (Pi) or when two ratios between Ca and Pi occur. Secondly, it was evaluated whether the ion absorption could be affected by the fatty acid pattern of enterocyte membrane. Results showed that in the absence of Pi, Ca transport from CaCl 2 was higher than from Ca-CPPs solutions. However, when Pi was added to the CaCl 2 solution, Ca absorption decreased by 90% (Ca:Pi = 1:1) and 97% (Ca:Pi = 1:2). On the contrary, when Pi was added to the Ca-CPPs solution, Ca transport decreased only by 40% (Ca:Pi = 1:1) and 60% (Ca:Pi = 1:2). The enrichment of enterocyte membrane with n-3 fatty acids (fish oil diet) compared to control diet did not modify the effect of CPPs on Ca transport in ileum of rats. These findings demonstrate that CPPs can limit the inhibitory effect of phosphate on Ca availability and increase Ca transport across the distal small intestine. This action could be of nutritional relevance especially when the dietary intake of this mineral is lower than the recommended level or there is an unbalanced ratio Ca/Pi in diet.

The effect of casein phosphopeptides on calcium absorption from calcium-fortified milk in growing rats.

The effect of casein phosphopeptides (CPP) prepared from bovine casein by enzymatic hydrolysis (extrinsic CPP) on Ca absorption from Ca-fortified milk was studied in young male rats, in comparison with that produced from casein in the small intestine (intrinsic CPP). The gastrointestinal Ca disappearance (Ca ingested - (gastric Ca + intestinal Ca )) was calculated as an indirect measurement of Ca absorption. After being fasted overnight, the animals were given 2.0 ml Ca-fortified milk (30 g fat, 35 g protein, 2.7 g Ca/kg) without or with 1.0 mg extrinsic CPP/ml, by gastric intubation. The intestinal soluble Ca level after 15 min and the gastrointestinal Ca disappearance after 15 and 30 min in the rats given Ca-fortified milk with 1.0 mg extrinsic CPP/ml were significantly higher than these figures in the rats given Ca-fortified milk without CPP (P < 0.05). When the rats were given unfortified milk (1.35 g Ca/kg) in another reference experiment, no significant effect on intestinal soluble Ca and gastrointestinal Ca disappearance was apparent from the addition of CPP to milk. Ca availability was estimated by measuring 45Ca-deposits in the bones of rats 48 h after being given 2.0 ml Ca-fortified milk labelled with 45Ca (180 kBq/2 ml) with or without 0.25 mg CPP/ml. The levels of 45Ca radioactivity of the femur and tibia from the rats given Ca-fortified milk with extrinsic CPP were significantly higher than those from the control group (P < 0.05). These results suggest that the addition of CPP to Ca-fortified milk could increase Ca absorption by growing rats mainly from CaCO3 added to the milk. The mechanism of CPP related to the interaction of CPP and Ca in the gastrointestinal tract is discussed.

The effect of casein phosphopeptides on zinc and calcium absorption from high phytate infant diets assessed in rat pups and Caco-2 cells.

Milk and other foods of animal origin have been shown to improve zinc absorption from phytate-rich diets. The ability of milk proteins and casein phosphopeptides (CPP), the latter formed during digestion of casein, to overcome the inhibitory effect of phytate on zinc and calcium absorption was investigated. Suckling rat pups were given aqueous phytate-containing solutions, oat diet, or soy formula alone or with milk proteins or CPP added. Diets labeled extrinsically with 65Zn and 47Ca were given by gastric intubation. Absorption was determined from measurement of radioisotope activity in intestine, organs, and carcass. Addition of CPP improved zinc and calcium absorption from aqueous phytate-containing solutions and from oat diet. The effect of CPP on calcium absorption from soy formula was less pronounced. The influence of CPP on zinc absorption from aqueous phytate-containing solutions was also examined using a human colon carcinoma-derived cell line, Caco-2. Binding + uptake of 65Zn was determined after incubation with these solutions. Phytate reduced zinc binding + uptake to 79% of the control value. Addition of 14 mumol of CPP/L increased zinc binding + uptake to 94%, whereas 36 and 72 mumol of CPP/L depressed zinc binding + uptake (75 and 39%). In conclusion, CPP improved zinc and calcium bioavailability from high phytate meals in the rat pup model. In the Caco-2 cell system, addition of 14 mumol of CPP/L showed a positive effect on zinc binding + uptake from phytate-containing solutions, whereas higher levels of CPP inhibited zinc binding + uptake.

Effect of casein phospho peptides and Ca2+ on penetration of boar spermatozoa into pig oocytes matured in vitro.

To investigate the promoting effect of casein phospho peptides (CPP) on the penetration of ejaculated boar spermatozoa into in vitro-matured oocytes, preincubated spermatozoa were coincubated with in vitro-matured oocytes in fertilization medium with or without CPP for 60-240 min. Sperm penetration into zona-intact oocytes was observed within 90 min of coincubation. The penetration rate reached a maximum earlier in the presence of CPP than in the absence of CPP. Furthermore, spermatozoa preincubated in fertilization medium with CPP before coincubation with matured oocytes retained the ability to penetrate oocytes for a longer time than spermatozoa preincubated in fertilization medium without CPP. In fertilization medium containing CPP, Ca2+ uptake by spermatozoa increased within 90 min of incubation, with a peak of 113 nm/1 x 10(7) cells. On the other hand, in fertilization medium with no CPP, the intracellular Ca2+ concentration remained constant or gradually decreased until 180 min of incubation. When preincubated spermatozoa were coincubated with oocytes for 5 h in a fertilization medium containing CPP and/or EGTA, or with no addition, EGTA decreased the rate of oocytes penetrated by spermatozoa and the number of spermatozoa penetrating each oocyte. On the other hand, when CPP was added to the medium containing EGTA, the inhibitory effect of EGTA was neutralized. These results indicate that CPP promotes Ca2+ uptake of boar spermatozoa, resulting in a more rapid and longer-lasting shift of a subpopulation of spermatozoa that can penetrate oocytes; the findings also suggest that CPP neutralizes the inhibitory effect of EGTA on sperm penetration.

Effect of caseinphosphopeptide on absorbability of co-ingested calcium in normal postmenopausal women

Caseinphosphopeptide in a dose of 87.5 mg was administered to 35 normal post-menopausal women as a part of a standard test meal containing a calcium load of 250 mg. Absorption of calcium was tested both with and without caseinphosphopeptide, using an intrinsic45Ca label in the calcium source. The mean quotient of absorption with/without caseinphosphopeptide was greater than 1.0, but nonsignificantly so. However, when analysis was confined to women with low absorption values, caseinphosphopeptide administration was associated with significantly better absorption of co-ingested calcium. Those findings suggest that caseinphosphopeptide supplementation is particularly useful, for persons with low basal absorptive performance.

Effect of casein phospho peptides on penetration of boar spermatozoa to oocytes matured in culture

We carried out the serotyping of 196 freshly isolated strains of Pseudomonas aeruginosa with international antigenic typing system (IATS) antibodies from Rougier Bio-Tech (RBT kit) and other two kinds of antibodies, from Meiji Seika Kaisha (Mei assay kit) and Denka Seiken (Seiken kit) to ascertain the relationship among the serotypes obtained by the different methods. All the test strains belonging to the serogroups A, C, E, F, G, H, I, J, and K were well correlated with each other with the Mei assay and the Seiken kits. However, 21 of 26 strains of serogroup M typed with the Mei assay kit were non-typable with the Seiken kit. It was confirmed that serogroups obtained with both these methods were closely related; 171 of the 196 strains of P. aeruginosa tested (87.2%) were classified into identical serogroups. From the serotyping results for P. aeruginosa strains with the Seiken and the RBT kits and with the Mei assay and the RBT kits, it was found that the serotypes of 172 of the 192 strains tested (89.6%) and those of 164 of the 170 strains tested (96.5%), respectively were well correlated. However, the serotyping of P. aeruginosa with the RBT kit may be problematic, as the kit did not contain the antisera agglutinated with the serogroup M strains of P. aeruginosa classified with the Mei assay and Seiken kits.

The influences of casein phosphopeptides on metabolism of ectopic bone induced by decalcified bone matrix implantation in rats.

The effect of casein phosphopeptides (CPP) on bone metabolism was studied in the ectopic bone induced by the implantation of decalcified bone matrix in rats. Forty-two Wistar male rats of 7 weeks old were fed low calcium diets (0.39% of calcium) with or without supplying 0.50% of CPP, or a control diet (0.91% of calcium) without CPP supplementation. After a 1-week preliminary period, each rat was subcutaneously implanted with 30 mg of demineralized bone matrix powder. Fourteen and 21 days after the implantation, the implants were harvested from 7 rats of each group. Calcium content in the graft was not significantly different among all groups on day 14. Subsequently, the content of calcium rapidly increased in the grafts irrespective of diets given. However, the graft of the CPP- group contained less calcium than the other groups and the calcium content was more in the control rats compared to the CPP+ animals on day 21. Alkaline phosphatase activity (an index of bone and cartilage calcification) was lower in the control group than in the CPP+ group on day 14. The enzyme activity subsequently decreased in the control group but the activity was not changed in the other groups. As a result, the activity of alkaline phosphatase was lower in the control animals than in the other rats on day 21. Tartrate-resistant cap phosphatase activity (an index of bone resorption) was higher in the CPP--group compared to the control on day 14. On day 21, the activity was higher in the CPP--group compared to the others.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of casein phosphopeptides and Ca 2+ on fertilization in vitro of bovine oocytes matured in culture

Effects of casein phosphopeptides and Ca 2+ on fertilization in vitro of bovine oocytes matured in culture

The effect of casein phosphopeptides on calcium utilization in young ovariectomized rats

The effect of casein phosphopeptides (CPPs) on Ca utilization in ovariectomized (OVX) rats was studied. A mixture of CPPs corresponding to the amino acid sequences 1-25 and 1-28 in the beta-casein was isolated from the tryptic digest of beta-casein (beta CPP). After being fed a low Ca diet for 30 days, OVX rats were fed experimental diets of which the Ca level was 0.1%, 0.3% or 0.5% with or without 0.15% beta CPP for 28 days. During days 1-3 of the Ca refeeding period, rats fed beta CPP with 0.5% Ca showed a higher Ca absorption than control rats not supplemented with beta CPP. During days 7-9 and 26-28, there were no significant differences in Ca and P balances between the beta CPP group and the control group for each dietary Ca level. Femoral Ca and P contents from rats fed beta CPP tended to be higher than those from control rats. These results suggest that beta CPP supplementation could have an effect on Ca absorption at a certain degree of Ca deficiency.

Casein phosphopeptides: their effect on calcification of in vitro cultured embryonic rat bone.

The effect of casein phosphopeptides (CPP) and enzymatically dephosphorylated, otherwise identical peptides (DPP) on Ca solubility and mineralization was studied in an in vitro model. The model consisted of explanted embryonic rat bone rudiments (femora, tibiae, and metatarsals), cultured in Bigger's medium at two different concentrations of total calcium (1.78 mM and 3.24 mM). The medium enabled growth of the explants, but minor or no mineralization took place during a culture period of 11 days. The medium was unstable with respect to Ca solubility, and measurements with a selective electrode showed that during most of the time the actual concentration of soluble, ionic Ca2+ was not higher than about 0.7 mM. CPP added at a concentration of 10 mg/100 ml of the medium vastly improved Ca-solubility under culture conditions. The mean concentration of soluble, ionic Ca2+ in the presence of the peptides was 1.4 mM at total calcium = 1.78 mM and 2.3 mM at the higher total calcium level (3.24 mM). Rudiments cultured in the presence of CPP showed impressive increases in calcification of their diaphyseal area. This progress in mineralization was quantitatively assayed by measuring the projected dark diaphyseal area at the beginning and at the end of the culture period. It was also verified by histological sections of the rudiments. DDP did not enhance Ca solubility of the culture medium, nor did explants cultured in the presence of 10 mg/100 ml of DPP show any significant progress in mineralization. This indicated that the activity of CPP was dependent on the presence of serine-linked phosphate groups.