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Sample records for caseins

  1. Radioimmunoassay of casein

    International Nuclear Information System (INIS)

    The heterogeneity of casein and its consequence on the radioimmunoassay are discussed. All protein fractions of human milk are chequed for cross reactivity. Casein K and II are labelled with 125I. The elution curves of these labelled compounds are measured by radioactivity and optical density. The sera of lactating women and of cancerous patients were chromatographed. In human milk a major peak is found in the elution region of casein. The serum of cancerous patients showed a heterogenous elution pattern

  2. Natural variation in casein composition of milk

    OpenAIRE

    van der Bijl, E.

    2014-01-01

    Bovine milk contains 3-4 % protein and almost 80% of the milk protein fraction consist of four caseins; αs1-casein, β-casein, αs2-casein and κ-casein. Most of the caseins in milk are assembled in casein micelles, which consist of several thousands of individual casein molecules and salts. The unique structure of casein micelles allows the delivery of large amounts of calcium and phosphate to the neonate. Considerable natural variation in casein content and composition e...

  3. Glycation Reactions of Casein Micelles.

    Science.gov (United States)

    Moeckel, Ulrike; Duerasch, Anja; Weiz, Alexander; Ruck, Michael; Henle, Thomas

    2016-04-13

    After suspensions of micellar casein or nonmicellar sodium caseinate had been heated, respectively, in the presence and absence of glucose for 0-4 h at 100 °C, glycation compounds were quantitated. The formation of Amadori products as indicators for the "early" Maillard reaction were in the same range for both micellar and nonmicellar caseins, indicating that reactive amino acid side chains within the micelles are accessible for glucose in a comparable way as in nonmicellar casein. Significant differences, however, were observed concerning the formation of the advanced glycation end products (AGEs), namely, N(ε)-carboxymethyllysine (CML), pyrraline, pentosidine, and glyoxal-lysine dimer (GOLD). CML could be observerd in higher amounts in nonmicellar casein, whereas in the micelles the pyrraline formation was increased. Pentosidine and GOLD were formed in comparable amounts. Furthermore, the extent of protein cross-linking was significantly higher in the glycated casein micelles than in the nonmicellar casein samples. Dynamic light scattering and scanning electron microscopy showed that glycation has no influence on the size of the casein micelles, indicating that cross-linking occurs only in the interior of the micelles, but altered the surface morphology. Studies on glycation and nonenzymatic cross-linking can contribute to the understanding of the structure of casein micelles. PMID:27018258

  4. REVIEW ON CASEIN PRODUCTION AND CASEIN BASED NANO-FORMULATIONS

    Directory of Open Access Journals (Sweden)

    Arora Neha

    2012-01-01

    Full Text Available Biodegradable systems have the ability to release the drug for a prolonged period of time and subsequently degrade which can be easily cleared from the body. This property makes use of them for the design of carriers for the controlled delivery of therapeutic agents, since it will release the entrapped drug over an extended period of time. Due to its non toxic, biocompatible and slow digesting nature it is an effective matrix in drug delivery. Milk has many proteins, including the nine essential amino acids. The two basic types of proteins in milk are called casein and whey. The ability of casein to modify drug dissolution from compacts was reported. The high tensile strength of casein film, favours its use as an acceptable film-coating for tablets. Casein floating beads were developed to increase the residence time of drugs in the stomach based on its emulsifying and bubble-forming properties. Casein-based microparticles bioactive molecules were prepared via emulsification-chemical cross linking Casein nanoformulations were also prepared to deliver nutraceuticals and synthetic drugs via enzymatic crosslinking, graft copolymerization, heat-gelation and polyelectrolyte ionic complexation. It can be concluded that casein-based formulations are promising materials for controlled drug delivery.

  5. Casein polymorphism heterogeneity influences casein micelle size in milk of individual cows.

    Science.gov (United States)

    Day, L; Williams, R P W; Otter, D; Augustin, M A

    2015-06-01

    Milk samples from individual cows producing small (148-155 nm) or large (177-222 nm) casein micelles were selected to investigate the relationship between the individual casein proteins, specifically κ- and β-casein phenotypes, and casein micelle size. Only κ-casein AA and β-casein A1A1, A1A2 and A2A2 phenotypes were found in the large casein micelle group. Among the small micelle group, both κ-casein and β-casein phenotypes were more diverse. κ-Casein AB was the dominant phenotype, and 3 combinations (AA, AB, and BB) were present in the small casein micelle group. A considerable mix of β-casein phenotypes was found, including B and I variants, which were only found in the small casein micelle group. The relative amount of κ-casein to total casein was significantly higher in the small micelle group, and the nonglycosylated and glycosylated κ-casein contents were higher in the milks with small casein micelles (primarily with κ-casein AB and BB variants) compared with the large micelle group. The ratio of glycosylated to nonglycosylated κ-casein was higher in the milks with small casein micelles compared with the milks with large casein micelles. This suggests that although the amount of κ-casein (both glycosylated and nonglycosylated) is associated with micelle size, an increased proportion of glycosylated κ-casein could be a more important and favorable factor for small micelle size. This suggests that the increased spatial requirement due to addition of the glycosyl group with increasing extent of glycosylation of κ-casein is one mechanism that controls casein micelle assembly and growth. In addition, increased electrostatic repulsion due to the sialyl residues on the glycosyl group could be a contributory factor. PMID:25828659

  6. Casein micelle structure: a concise review

    OpenAIRE

    Chanokphat Phadungath

    2005-01-01

    Milk is a complex biological fluid with high amount of proteins, lipid and minerals. The function of milk is to supply nutrients such as essential amino acids required for the growth of the newborn. In addition, due to the importance of casein and casein micelles for the functional behavior of dairy products, the nature and structure of casein micelles have been studied extensively. However, the exact structure of casein micelles is still under debate. Various models for casein micelle struct...

  7. PROTEIN-PROTEIN INTERACTIONS IN CASEIN PRECIPITATED BY PRESSURIZED CARBON DIOXIDE (C02 CASEIN)

    Science.gov (United States)

    Despite much research on the casein micelles, the colloidal calcium-phosphate complexes of skim milk, the molecular interactions involved in the casein micelles remain elusive. We investigate casein precipitated by pressurized carbon dioxide (CO2 casein) to elucidate the mechanism of the formation o...

  8. Solubilization of genistein by caseinate micellar system.

    Science.gov (United States)

    Anjani, Gemala; Ohta, Akio; Yasuhara, Kazuma; Asakawa, Tsuyoshi

    2014-01-01

    This study investigates the aggregation behavior of caseinate and the solubilization of genistein in aqueous caseinate solution. The critical aggregation concentration (CAC) of caseinate was obtained from the fluorescence intensity of 8-anilino-1-naphthalenesulfonic acid (ANS), which was enhanced by ANS-protein interactions and the hydrophobicity of caseinate. The increasing solubility of genistein in caseinate was confirmed by HPLC measurements; above and below the CAC, the genistein/caseinate molar ratio is 1:1 and 10:1, respectively. The latter ratio indicates that more caseinate molecules surround genistein below the CAC. However, the solubility of genistein in caseinate is unaffected by calcium ions. Atomic force microscopy (AFM) shows that casein sub-micelles are similarly structured in the presence and absence of genistein. In AFM phase images, the caseinate sub-micelle is brightened in the presence of genistein, implying that the particle becomes more rigid, probably because genistein attaches to the surface or to the narrow part of the sub-micelle. The diameter of sub-micelle aggregates is two times that of caseinate alone (24 nm versus 12 nm). These results were confirmed by cryo-TEM observations. PMID:24599106

  9. Development of casein microgels from cross-linking of casein micelles by genipin.

    Science.gov (United States)

    Silva, Naaman F Nogueira; Saint-Jalmes, Arnaud; de Carvalho, Antônio F; Gaucheron, Frédéric

    2014-09-01

    Casein micelles are porous colloidal particles, constituted of casein molecules, water, and minerals. The vulnerability of the supramolecular structure of casein micelles face to changes in the environmental conditions restrains their applications in other domains besides food. Thus, redesigning casein micelles is a challenge to create new functionalities for these biosourced particles. The objective of this work was to create stable casein microgels from casein micelles using a natural cross-linker, named genipin. Suspensions of purified casein micelles (25 g L(-1)) were mixed with genipin solutions to have final concentrations of 5, 10, and 20 mM genipin. Covalently linked casein microgels were formed via cross-linking of lysyl and arginyl residues of casein molecules. The reacted products exhibited blue color. The cross-linking reaction induced gradual changes on the colloidal properties of the particles. The casein microgels were smaller and more negatively charged and presented smoother surfaces than casein micelles. These results were explained based on the cross-linking of free NH2 present in an external layer of κ-casein. Light scattering and rheological measurements showed that the reaction between genipin and casein molecules was intramicellar, as one single population of particles was observed and the values of viscosity (and, consequently, the volume fraction of the particles) were reduced. Contrary to the casein micelles, the casein microgels were resistant to the presence of dissociating agents, e.g., citrate (calcium chelating) and urea, but swelled as a consequence of internal electrostatic repulsion and the disruption of hydrophobic interactions between protein chains. The casein microgels did not dissociate at the air-solution interface and formed solid-like interfaces rather than a viscoelastic gel. The potential use of casein microgels as adaptable nanocarriers is proposed in the article. PMID:25117401

  10. Genetic polymorphism of kappa casein and casein micelle size in the Bulgarian Rhodopean cattle breed

    OpenAIRE

    Hristov P.; Neov B.; Sbirkova H.; Teofanova D.; Radoslavov G.; Shivachev B.

    2014-01-01

    The present study aimed to compare the size of casein micelle in cow milk sample in function of kappa casein (CSN3) genetic polymorphism. Sixteen cows from Bulgarian Rhodopean cattle breed were genotyped by PCRRFLP analysis. Milk samples from the three found CSN3 genotypes (AB, AA and BB) were employed for the determination of casein micelles size by Dynamic Light Scattering (DLS). The results showed differences in the size and polydispersity of the casein ...

  11. Electrosorption of pectin onto casein micelles

    OpenAIRE

    Tuinier, R.; Rolin, C.; de Kruif, C.G.

    2002-01-01

    Pectin, a polysaccharide derived from plant cells of fruit, is commonly used as stabilizer in acidified milk drinks. To gain a better understanding of the way that pectin stabilizes these drinks, we studied the adsorption and layer thickness of pectin on casein micelles in skim milk dispersions. Dynamic light scattering was used to measure the layer thickness of adsorbed pectin onto casein micelles in situ during acidification. The results indicate that the adsorption of pectin onto casein mi...

  12. Casein Micelle Dispersions under Osmotic Stress

    OpenAIRE

    Bouchoux, Antoine; Cayemitte, Pierre-Emerson; Jardin, Julien; Gésan-Guiziou, Geneviève; Cabane, Bernard

    2009-01-01

    Casein micelles dispersions have been concentrated and equilibrated at different osmotic pressures using equilibrium dialysis. This technique measured an equation of state of the dispersions over a wide range of pressures and concentrations and at different ionic strengths. Three regimes were found. i), A dilute regime in which the osmotic pressure is proportional to the casein concentration. In this regime, the casein micelles are well separated and rarely interact, whereas the osmotic press...

  13. Casein genetic polymorphisms in goat breeds of Lombardy

    Directory of Open Access Journals (Sweden)

    G. Pagnacco

    2010-01-01

    Full Text Available In the last years, the genetic polymorphism of goat caseins has raised a considerable research interest due to relationships with milk quality and technological properties (Martin et al., 2002. The four caseins, αs1-casein (CSN1S1, β-casein (CSN2, αs2-casein (CSN1S2, and k-casein (CSN3, are coded by a cluster of genes (Ferretti et al., 1990; Threadgill and Womack, 1990.

  14. Casein genetic polymorphisms in goat breeds of Lombardy

    OpenAIRE

    G. Pagnacco; A. Caroli; P. Bolla; F. Chiatti; Chessa, S.

    2010-01-01

    In the last years, the genetic polymorphism of goat caseins has raised a considerable research interest due to relationships with milk quality and technological properties (Martin et al., 2002). The four caseins, αs1-casein (CSN1S1), β-casein (CSN2), αs2-casein (CSN1S2), and k-casein (CSN3), are coded by a cluster of genes (Ferretti et al., 1990; Threadgill and Womack, 1990).

  15. Milk composition and lactation of beta-casein-deficient mice.

    OpenAIRE

    Kumar, S.; Clarke, A R; Hooper, M L; Horne, D S; Law, A J; Leaver, J; Springbett, A; Stevenson, E.; Simons, J.P.

    1994-01-01

    beta-Casein is a major protein component of milk and, in conjunction with the other caseins, it is assembled into micelles. The casein micelles determine many of the physical characteristics of milk, which are important for stability during storage and for milk-processing properties. There is evidence that suggests that beta-casein may also possess other, nonnutritional functions. To address the function of beta-casein, the mouse beta-casein gene was disrupted by gene targeting in embryonic s...

  16. Coacervates of lysozyme and β-casein

    NARCIS (Netherlands)

    Anema, S.G.; de Kruif, C.G.

    2013-01-01

    Complexes are formed when positively charged lysozyme (LYZ) is mixed with negatively charged caseins. Adding b-casein (BCN) to LYZ leads to flocculation even at low addition levels. Titrating LYZ into BCN shows that complexes are formed up to a critical composition (x = [LYZ]/([LYZ] + [BCN]). The fo

  17. Milk Intolerance, Beta-Casein and Lactose

    Directory of Open Access Journals (Sweden)

    Sebely Pal

    2015-08-01

    Full Text Available True lactose intolerance (symptoms stemming from lactose malabsorption is less common than is widely perceived, and should be viewed as just one potential cause of cows’ milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows’ milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows’ milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

  18. Milk Intolerance, Beta-Casein and Lactose.

    Science.gov (United States)

    Pal, Sebely; Woodford, Keith; Kukuljan, Sonja; Ho, Suleen

    2015-09-01

    True lactose intolerance (symptoms stemming from lactose malabsorption) is less common than is widely perceived, and should be viewed as just one potential cause of cows' milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows' milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows' milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed. PMID:26404362

  19. Characterisation of monotreme caseins reveals lineage-specific expansion of an ancestral casein locus in mammals.

    Science.gov (United States)

    Lefèvre, Christophe M; Sharp, Julie A; Nicholas, Kevin R

    2009-01-01

    Using a milk-cell cDNA sequencing approach we characterised milk-protein sequences from two monotreme species, platypus (Ornithorhynchus anatinus) and echidna (Tachyglossus aculeatus) and found a full set of caseins and casein variants. The genomic organisation of the platypus casein locus is compared with other mammalian genomes, including the marsupial opossum and several eutherians. Physical linkage of casein genes has been seen in the casein loci of all mammalian genomes examined and we confirm that this is also observed in platypus. However, we show that a recent duplication of beta-casein occurred in the monotreme lineage, as opposed to more ancient duplications of alpha-casein in the eutherian lineage, while marsupials possess only single copies of alpha- and beta-caseins. Despite this variability, the close proximity of the main alpha- and beta-casein genes in an inverted tail-tail orientation and the relative orientation of the more distant kappa-casein genes are similar in all mammalian genome sequences so far available. Overall, the conservation of the genomic organisation of the caseins indicates the early, pre-monotreme development of the fundamental role of caseins during lactation. In contrast, the lineage-specific gene duplications that have occurred within the casein locus of monotremes and eutherians but not marsupials, which may have lost part of the ancestral casein locus, emphasises the independent selection on milk provision strategies to the young, most likely linked to different developmental strategies. The monotremes therefore provide insight into the ancestral drivers for lactation and how these have adapted in different lineages. PMID:19874726

  20. Casein micelle structure: a concise review

    Directory of Open Access Journals (Sweden)

    Chanokphat Phadungath

    2005-01-01

    Full Text Available Milk is a complex biological fluid with high amount of proteins, lipid and minerals. The function of milk is to supply nutrients such as essential amino acids required for the growth of the newborn. In addition, due to the importance of casein and casein micelles for the functional behavior of dairy products, the nature and structure of casein micelles have been studied extensively. However, the exact structure of casein micelles is still under debate. Various models for casein micelle structure have been proposed. Most of the proposedmodels fall into three general categories, which are: coat-core, subunit (sub-micelles, and internal structure models. The coat-core models, proposed by Waugh and Nobel in 1965, Payens in 1966, Parry and Carroll in 1969, and Paquin and co-workers in 1987, describe the micelle as an aggregate of caseins with outer layer differing in composition form the interior, and the structure of the inner part is not accurately identified. The sub-micelle models, proposed by Morr in 1967, Slattery and Evard in 1973, Schmidt in 1980, Walstra in1984, and Ono and Obata in 1989, is considered to be composed of roughly spherical uniform subunits. The last models, the internal structure models, which were proposed by Rose in 1969, Garnier and Ribadeau- Dumas in 1970, Holt in 1992, and Horne in 1998, specify the mode of aggregation of the different caseins.

  1. Milk Intolerance, Beta-Casein and Lactose

    OpenAIRE

    Sebely Pal; Keith Woodford; Sonja Kukuljan; Suleen Ho

    2015-01-01

    True lactose intolerance (symptoms stemming from lactose malabsorption) is less common than is widely perceived, and should be viewed as just one potential cause of cows’ milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows’ milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-...

  2. Casein expression in cytotoxic T lymphocytes.

    OpenAIRE

    Grusby, M J; Mitchell, S C; Nabavi, N; Glimcher, L H

    1990-01-01

    A cDNA that expresses a mRNA restricted to cytotoxic T lymphocytes (CTL) and mammary tissue has been isolated and characterized. The deduced amino acid sequence from this cDNA shows extensive homology with the previously reported amino acid sequence for rat alpha-casein. Indeed, the presence of a six-residue-repeated motif that is specific for rodent alpha-caseins strongly supports the identification of this cDNA as mouse alpha-casein. Northern (RNA) blot analysis of many hematopoietic cell t...

  3. Structural changes of casein micelles in a calcium gradient film

    OpenAIRE

    Gebhardt, R.; Burghammer, M.; Riekel, C.; Roth, S. V.; Müller-Buschbaum, P.

    2008-01-01

    Calcium gradients are prepared by sequentially filling a micropipette with casein solutions of varying calcium concentration and spreading them on glass slides. The casein film is formed by a solution casting process, which results in a macroscopically rough surface. Microbeam grazing incidence small-angle X-ray scattering (microGISAXS) is used to investigate the lateral size distribution of three main components in casein films: casein micelles, casein mini-micelles, and micellar calcium pho...

  4. COMPARATIVE STUDY ON MILK CASEIN ASSAY METHODS

    Directory of Open Access Journals (Sweden)

    RODICA CĂPRIłĂ

    2013-12-01

    Full Text Available Casein, the main milk protein was determined by different assay methods: the gravimetric method, the method based on the neutralization of the NaOH excess used for the casein precipitate solving and the method based on the titration of the acetic acid used for the casein precipitation. The last method is the simplest one, with the fewer steps, and also with the lowest error degree. The results of the experiment revealed that the percentage of casein from the whole milk protein represents between 72.6–81.3% in experiment 1, between 73.6–81.3% in experiment 2 and between 74.3–81% in experiment 3.

  5. Interactions of casein micelles with calcium phosphate particles.

    Science.gov (United States)

    Tercinier, Lucile; Ye, Aiqian; Anema, Skelte G; Singh, Anne; Singh, Harjinder

    2014-06-25

    Insoluble calcium phosphate particles, such as hydroxyapatite (HA), are often used in calcium-fortified milks as they are considered to be chemically unreactive. However, this study showed that there was an interaction between the casein micelles in milk and HA particles. The caseins in milk were shown to bind to the HA particles, with the relative proportions of bound β-casein, αS-casein, and κ-casein different from the proportions of the individual caseins present in milk. Transmission electron microscopy showed no evidence of intact casein micelles on the surface of the HA particles, which suggested that the casein micelles dissociated either before or during binding. The HA particles behaved as ion chelators, with the ability to bind the ions contained in the milk serum phase. Consequently, the depletion of the serum minerals disrupted the milk mineral equilibrium, resulting in dissociation of the casein micelles in milk. PMID:24896851

  6. RECONSTITUTED MICELLE FORMATION USING REDUCED, CARBOXYMETHYLATED BOVINE K-CASEIN AND HUMAN B-CASEIN

    Science.gov (United States)

    In milk, K-casein, a mixture of disulfide-bonded polymers, stabilizes and regulates the size of the unique colloidal complex of protein, Ca+2 and inorganic phosphate (Pi) termed the casein (CN) micelle. However, reduced, carboxymethylated bovine K-CN (RCM-K) forms fibrils at 37 degrees C and its mi...

  7. Casein micelles and their internal structure

    Energy Technology Data Exchange (ETDEWEB)

    De Kruif, Cornelis G [ORNL; Huppertz, Thom [NIZO Food Research; Urban, Volker S [ORNL; Petukhov, Andrei V [Van ' t Hoff laboratory for Physical and Colloid Chemistry, Utrecht University, The Netherlands

    2012-01-01

    The internal structure of casein micelles was studied by calculating the small-angle neutron and X-ray scattering and static light scattering spectrum (SANS, SAXS, SLS) as a function of the scattering contrast and composition. We predicted experimental SANS, SAXS, SLS spectra self consistently using independently determined parameters for composition size, polydispersity, density and voluminosity. The internal structure of the casein micelles, i.e. how the various components are distributed within the casein micelle, was modeled according to three different models advocated in the literature; i.e. the classical sub-micelle model, the nanocluster model and the dual binding model. In this paper we present the essential features of these models and combine new and old experimental SANS, SAXS, SLS and DLS scattering data with new calculations that predict the spectra. Further evidence on micellar substructure was obtained by internally cross linking the casein micelles using transglutaminase, which led to casein nanogel particles. In contrast to native casein micelles, the nanogel particles were stable in 6 M urea and after sequestering the calcium using trisodium citrate. The changed scattering properties were again predicted self consistently. An important result is that the radius of gyration is independent of contrast, indicating that the mass distribution within a casein micelle is homogeneous. Experimental contrast is predicted quite well leading to a match point at a D{sub 2}O volume fraction of 0.41 ratio in SANS. Using SANS and SAXS model calculations it is concluded that only the nanocluster model is capable of accounting for the experimental scattering contrast variation data. All features and trends are predicted self consistently, among which the 'famous' shoulder at a wave vector value Q = 0.35 nm{sup -1}. In the nanocluster model, the casein micelle is considered as a (homogeneous) matrix of caseins in which the colloidal calcium phosphate (CCP

  8. Casein fraction of ovine milk from indigenous Greek breeds

    OpenAIRE

    Moatsou, Golfo; Samolada, Maria; Katsabeki, Alexandra; Anifantakis, Emmanuel

    2004-01-01

    Whole casein fractions isoelectrically prepared from bulk milks from four different indigenous Greek ovine breeds were analysed by urea-PAGE and reversed-phase HPLC. Individual caseins prepared by fractionation on a cation-exchange column were used to locate the peaks on the chromatograms. Apart from heterogeneity that was depicted in the peak shape of caseins, there was also quantitative variability regarding the $\\alpha$s- and $\\beta$-casein contents. According to the RP-HPLC results, the r...

  9. hermal decomposition of irradiated casein molecules

    International Nuclear Information System (INIS)

    NON-Isothermal studies were carried out using the derivatograph where thermogravimetry (TG) and differential thermogravimetry (DTG) measurements were used to obtain the activation energies of the first and second reactions for casein (glyco-phospho-protein) decomposition before and after exposure to 1 Gy γ-rays and up to 40 x 104 μg Gy fast neutrons. 25Cf was used as a source of fast neutrons, associated with γ-rays. 137Cs source was used as pure γ-source. The activation energies for the first and second reactions for casein decomposition were found to be smaller at 400 μGy than that at lower and higher fast neutron doses. However, no change in activation energies was observed after γ-irradiation. it is concluded from the present study that destruction of casein molecules by low level fast neutron doses may lead to changes of shelf storage period of milk

  10. Effects of hydrolysed casein, intact casein and intact whey protein on energy expenditure and appetite regulation

    DEFF Research Database (Denmark)

    Bendtsen, Line Quist; Lorenzen, Janne Kunchel; Gomes, Sisse;

    2014-01-01

    Casein and whey differ in amino acid composition and in the rate of absorption; however, the absorption rate of casein can be increased to mimic that of whey by exogenous hydrolysis. The objective of the present study was to compare the effects of hydrolysed casein (HC), intact casein (IC) and...... intact whey (IW) on energy expenditure (EE) and appetite regulation, and thereby to investigate the influence of amino acid composition and the rate of absorption. In the present randomised cross-over study, twenty-four overweight and moderately obese young men and women consumed three isoenergetic...... dietary treatments that varied in protein source. The study was conducted in a respiration chamber, where EE, substrate oxidation and subjective appetite were measured over 24 h at three independent visits. Moreover, blood and urine samples were collected from the participants. The results showed no...

  11. A sensitive radioimmunoassay for a component of mouse casein

    International Nuclear Information System (INIS)

    Mouse casein (m.w. 22,000 daltons) has been purified by employing Sephadex G-100 and DEAE-cellulose column chromatographies. A sensitive radioimmunoassay method has been developed by using [125I]-labelled casein and antiserum elicited in rabbits after injection of glutaraldehyde-treated casein. The assay method is capable of detecting as little as 0.1 ng of casein. The use of the present radioimmunoassay method in detecting casein production in cultured mouse mammary explants has also been demonstrated

  12. Co-acervates of lactoferrin and caseins

    NARCIS (Netherlands)

    Anema, S.G.; de Kruif, C.G.

    2012-01-01

    On mixing positively charged lactoferrin (LF) with negatively charged caseins (*CN) it is observed that complexes are formed. The * stands for α, β, κ or Na. The size of the complex co-acervates appears to grow indefinitely and asymptotically near the point of charge equivalency. Away from the charg

  13. Casein kinase-2 structure-function relationship

    DEFF Research Database (Denmark)

    Boldyreff, B; Meggio, F; Pinna, L A;

    1992-01-01

    Nine mutants of human casein kinase-2 beta subunit have been created and assayed for their ability to assemble with the catalytic alpha subunit to give, at a 1:1 molar ratio, a fully competent CK-2 holoenzyme as judged by the following criteria: 1) the generation of an active heterotetrameric for...

  14. Casein - whey protein interactions in heated milk

    NARCIS (Netherlands)

    Vasbinder, Astrid Jolanda

    2003-01-01

    Heating of milk is an essential step in the processing of various dairy products, like for example yoghurt. A major consequence of the heat treatment is the denaturation of whey proteins, which either associate with the casein micelle or form soluble whey protein aggregates. By combination of enzyma

  15. αS1-casein, which is essential for efficient ER-to-Golgi casein transport, is also present in a tightly membrane-associated form

    OpenAIRE

    Le Parc, Annabelle; LEONIL, Joëlle; Chanat, Eric

    2010-01-01

    Background Caseins, the main milk proteins, aggregate in the secretory pathway of mammary epithelial cells into large supramolecular structures, casein micelles. The role of individual caseins in this process and the mesostructure of the casein micelle are poorly known. Results In this study, we investigate primary steps of casein micelle formation in rough endoplasmic reticulum-derived vesicles prepared from rat or goat mammary tissues. The majority of both αS1- and β-casein which are cystei...

  16. AlphaS1-casein, which is essential for efficient ER-to-Golgi casein transport, is also present in a tightly membrane-associated form

    OpenAIRE

    Le Parc, Annabelle; Léonil, Joelle; Chanat, Eric

    2010-01-01

    BACKGROUND: Caseins, the main milk proteins, aggregate in the secretory pathway of mammary epithelial cells into large supramolecular structures, casein micelles. The role of individual caseins in this process and the mesostructure of the casein micelle are poorly known. RESULTS: In this study, we investigate primary steps of casein micelle formation in rough endoplasmic reticulum-derived vesicles prepared from rat or goat mammary tissues. The majority of both alphaS1- and beta-casein which a...

  17. Interaction of lactoferrin and lysozyme with casein micelles.

    Science.gov (United States)

    Anema, Skelte G; de Kruif, C G Kees

    2011-11-14

    On addition of lactoferrin (LF) to skim milk, the turbidity decreases. The basic protein binds to the caseins in the casein micelles, which is then followed by a (partial) disintegration of the casein micelles. The amount of LF initially binding to casein micelles follows a Langmuir adsorption isotherm. The kinetics of the binding of LF could be described by first-order kinetics and similarly the disintegration kinetics. The disintegration was, however, about 10 times slower than the initial adsorption, which allowed investigating both phenomena. Kinetic data were also obtained from turbidity measurements, and all data could be described with one equation. The disintegration of the casein micelles was further characterized by an activation energy of 52 kJ/mol. The initial increase in hydrodynamic size of the casein micelles could be accounted for by assuming that it would go as the cube root of the mass using the adsorption and disintegration kinetics as determined from gel electrophoresis. The results show that LF binds to casein micelles and that subsequently the casein micelles partly disintegrate. All micelles behave in a similar manner as average particle size decreases. Lysozyme also bound to the casein micelles, and this binding followed a Langmuir adsorption isotherm. However, lysozyme did not cause the disintegration of the casein micelles. PMID:21932853

  18. Casein - whey protein interactions in heated milk

    OpenAIRE

    Vasbinder, Astrid Jolanda

    2003-01-01

    Heating of milk is an essential step in the processing of various dairy products, like for example yoghurt. A major consequence of the heat treatment is the denaturation of whey proteins, which either associate with the casein micelle or form soluble whey protein aggregates. By combination of enzymatic fractionation and capillary electrophoresis we were able to quantitatively determine the distribution of denatured whey proteins after heat treatment. This thesis describes the relation between...

  19. Thermal decomposition of irradiated casein molecules

    International Nuclear Information System (INIS)

    Non-isothermal studies were carried out using the derivatograph where thermogravimetry (TG), and differential thermogravimetry (DTG) measurements were used to obtain the activation energies of the first and second reactions for casein decomposition before and after exposure to gamma rays and fast neutrons. Cf- 252 was used as a source of fast neutrons associated with gamma rays. TG and DTG patterns were also recorded for casein samples before and after irradiation with 1 Gy gamma-rays of 0.662 MeV from Cs - 137. However, no change in a activation energies were observed after exposure to gamma-irradiation. On the other hand, the activation energies for first and second reactions were found to be smaller at 0.4 m Gy than that at lower and higher neutron doses. However, no change in activation energies was observed after γ irradiation. It is concluded from the present study that destruction of casein molecules by low level fast neutron doses may lead to changes of shelf storage period milk. 3 figs., 1 tab

  20. Disorder in milk proteins: caseins, intrinsically disordered colloids.

    Science.gov (United States)

    Redwan, Elrashdy M; Xue, Bin; Almehdar, Hussein A; Uversky, Vladimir N

    2015-01-01

    This article opens a series of reviews on the abundance and roles of intrinsic disorder in milk proteins. The focus of this introductory article on caseins is symbolic, since caseins were among the first recognized functional unfolded proteins and since they are definitely the most disordered, the most abundant, and the most studied of all milk proteins. In eutherian milks, the casein family includes at least three and usually four major members (αs1-, αs2-, β-, and κ-caseins) that are unrelated in sequence. However, in some species, two different αS2-casein genes are active, and therefore the total number of caseins can be as high as five. These proteins have found a number of uses in food industry. The functional repertoire of caseins ranges from nutritional function to involvement in the improving and/or maintaining cardiovascular health, to crucial contribution to the milk capacity to transport calcium phosphate, to serve as molecular chaperones, and to protect the mother's mammary gland against amyloidoses and ectopic calcification. An intricate feature of caseins is their ability to assemble to colloidal protein particles, casein micelles, serving to sequester and transport amorphous calcium phosphate. These and many other functions of caseins are obviously dependent on their intrinsically disordered nature and are controlled by various posttranslational modifications. Since various aspects of casein structure and function are rather well studied and since several recent reviews emphasized the functional roles of caseins' intrinsic disorder, the major goal of this article is to show how intrinsic disorder is encoded in the amino acid sequences of these proteins. PMID:25714333

  1. Long range restriction analysis of the bovine casein genes.

    OpenAIRE

    Ferretti, L; Leone, P.; Sgaramella, V

    1990-01-01

    Pulsed field gel electrophoresis (PFGE) was used to analyse the organization of the bovine alpha s1, alpha s2, beta and kappa casein genes. High molecular weight DNA was prepared from fibroblasts and lymphocytes embedded in agarose and was digested with the restriction endonucleases Clal, Sall, Smal, Xhol. The digestion products were separated by PFGE, transfered to nitrocellulose filters and hybridized to probes corresponding to the cDNAs of the four bovine casein genes. The casein genes wer...

  2. Characterization of casein and Poly-L-arginine Multilayer Films

    OpenAIRE

    Warszynska, Lilianna Szyk; Kilan, Katarzyna; Socha, Robert P.

    2013-01-01

    Thin films containing casein appear to be a promising material for coatings used in the medical area to promote biomineralization. alfa- and beta-casein and poly-L-arginine multilayer films were formed by the layer-by layer technique and their thickness and mass were analyzed by ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D). We investigated the effect of the type of casein used for the film formation and of the polyethyleneimine anchoring layer on the thickn...

  3. Casein genes of Bos taurus. II. Isolation and characterization of the β-casein gene

    International Nuclear Information System (INIS)

    The expression of the casein genes in the cells of the mammary gland is regulated by peptide and steroid hormones. In order to study the controlling mechanisms we have isolated and characterized the β-casein gene. The gene is 8.6 kb long and exceeds by a factor of 7.8 the length of the corresponding mRNA which is encoded by nine exons. The genomic clones incorporate in addition 8.5 kb and 4.5 kb of the 5'- and 3'-flanking regions. We have determined the sequence of the 5- and 3-terminals of the gene and have performed a comparative analysis of the corresponding regions of the rat β-casein gene. Furthermore we have identified the conversed sequences identical or homologous to the potential sections of binding to the nuclear factor CTF/NF-1 by glucocorticoid and progesterone receptors. The regulatory region of the bovine casein gene contains two variants of the TATA signal, flanking the duplication section in the promoter region

  4. Co-acervates of lactoferrin and caseins

    OpenAIRE

    Anema, S.G.; Kruif, C.G. de

    2012-01-01

    On mixing positively charged lactoferrin (LF) with negatively charged caseins (*CN) it is observed that complexes are formed. The * stands for α, β, κ or Na. The size of the complex co-acervates appears to grow indefinitely and asymptotically near the point of charge equivalency. Away from the charge equivalent ratio it seems that build-up of (surface) charges limits complex size. We proposed a simple scaling law so as to predict the size of the complex. By assuming that surface charge densit...

  5. Water holding capacity and swelling of casein hydrogels

    NARCIS (Netherlands)

    de Kruif, C; Anema, Skelte G.; Zhu, Changjun; Havea, Palatasa; Coker, Christina

    2015-01-01

    The water holding capacity of casein gels was investigated by measuring the swelling and de-swelling under a variety of conditions of temperature and salt concentration. Transglutaminase cross-linked sodium caseinate (15% w/w) gels will swell in good solvents or shrink in poor solvents until an equi

  6. 21 CFR 520.1157 - Iodinated casein tablets.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Iodinated casein tablets. 520.1157 Section 520... tablets. (a) Specifications. Each 1-gram tablet contains 25 milligrams of iodinated casein. (b) Sponsor. See No. 017762 in § 510.600(c) of this chapter. (c) Conditions of use—(1) Amount. 1/5 to 1 tablet...

  7. Is there a feeding strategy to increase milk casein content?

    Directory of Open Access Journals (Sweden)

    A. Formigoni

    2010-04-01

    Full Text Available Because more than 60% of milk produced in Italy is transformed into cheese, milk economical value strongly depends on cheese yield. Among the factors that influence cheese yield, milk casein and fat content plays a major role: when milk is converted into Grana Padano and Parmigiano reggiano, three grams of seasoned cheese are produced from one gram of milk casein.....

  8. Influence of succinylation on physicochemical property of yak casein micelles.

    Science.gov (United States)

    Yang, Min; Yang, Jitao; Zhang, Yuan; Zhang, Weibing

    2016-01-01

    Succinylation is a chemical-modification method that affects the physicochemical characteristics and functional properties of proteins. This study assessed the influence of succinylation on the physicochemical properties of yak casein micelles. The results revealed that surface hydrophobicity indices decreased with succinylation. Additionally, denaturation temperature and denaturation enthalpy decreased with increasing succinylation level, except at 82%. The buffering properties of yak casein micelles were affected by succinylation. It was found that chemical modification contributed to a slight shift of the buffering peak towards a lower pH value and a markedly increase of the maximum buffering values of yak casein micelles at pH 4.5-6.0 and pH casein micellar hydration and whiteness values. The findings obtained from this study will provide the basic information on the physicochemical properties of native and succinylated yak casein micelles. PMID:26213046

  9. Influence of succinylation on the conformation of yak casein micelles.

    Science.gov (United States)

    Yang, Min; Cui, Na; Fang, Yan; Shi, Ying; Yang, Jitao; Wang, Jiangyu

    2015-07-15

    Succinylation modifies the physicochemical characteristics and improves the functional properties of proteins. This study assessed the effects of succinylation on the conformation of yak casein micelles with seven degree of modification. The results revealed that succinylation contributed to the dissociation of casein micelles. With the increase of succinylated degree, soluble nitrogen and minerals content increased, while casein micelle size and polydispersity index of micelles decreased. Succinylation affected the spatial conformation of yak casein micelles: turn decreased, ß-sheet and α-helix increased, and irregular structure were non-significantly affected. The intrinsic and ANS fluorescence intensity decreased and the maximum emission wavelength shifted red with increasing succinylation. Based on the results, the structure of yak casein micelles was characteristic of the sub-micelle model. PMID:25722161

  10. Lipid oxidation in algae oil-in-water emulsions stabilized by bovine and caprine caseins

    Science.gov (United States)

    Caseins (alpha S1-, alpha S2-, and beta-casein) are phosphoproteins that are capable of binding transition metals and scavenging free radicals, these properties make them good candidates to be used as natural antioxidants in oil-in-water emulsions. Caprine casein exhibits variability in aS1-casein c...

  11. Casein Phosphopeptide- Amorphous Calcium Phosphate in Dentistry: A Literature Review

    OpenAIRE

    KESKİN, Arş. Gör. Dt. Gül; GÜLER, Yrd. Doç. Dr. Çiğdem

    2013-01-01

    Casein is a phosphoprotein in bovine milk and accounts for almost 80 percent of its total protein. Casein fosfopeptit (CPP) is produced as a result of decomposition of casein with trypsin enzyme using a selective deposition method. CPP can remarkably stabilize calcium phosphate in a state-forming CPPamorphous calcium phosphate (ACP) complex. CPPACP is located in topical gels, sugar-free chewing gum and mint dragees at the present day. In this paper, the effectiveness of CPP-ACP and usage in d...

  12. Alginate-Casein Microspheres as Bioactive Vehicles for Nutrients

    Institute of Scientific and Technical Information of China (English)

    何志敏; 张茜青; 齐崴; 黄仁亮; 苏荣欣

    2015-01-01

    The aim of this work was to develop an alginate-casein composite microsphere as a bioactive vehicle for oral administration of nutrients by a simple extrusion dripping method. Riboflavin was selected as a model drug, and the microencapsulation efficiency was raised to 97.94%after optimizing the preparation conditions by response surface methodology. In vitro release studies showed that riboflavin was released completely from alginate-casein microspheres in simulated intestinal fluids. Meanwhile, the morphology, structure and interaction between alginate and casein were characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectra.

  13. Process standardization for rennet casein based Mozzarella cheese analogue

    OpenAIRE

    Shah, Rahul; Jana, Atanu H.; Aparnathi, K. D.; Prajapati, P. S.

    2010-01-01

    A process for manufacture of Mozzarella cheese analogue (MCA) using rennet casein and plastic cream as protein and fat sources respectively was standardized. The formulation comprised of 25% plastic cream (72% fat), 27% rennet casein along with 3% tri-sodium citrate as emulsifying salt, 2% maltodextrin as binder, 0.55% lactic acid as pH regulator, 1% common salt for seasoning, 1% Mozzarella cheese bud as flavouring and 40.4% water. The process involved (a) dissolving the dry mixture of casein...

  14. Nucleolin (C23), a physiological substrate for casein kinase II

    DEFF Research Database (Denmark)

    Schneider, H R; Issinger, O G

    1988-01-01

    Nucleolin (C23), a 110 kDa phosphoprotein, which is mainly found in the nucleolus has been shown to be a physiological substrate for casein kinase II (CKII). Nucleolin was identified and characterized by immunodetection using an anti-nucleolin antibody. Phosphopeptide patterns from nucleolin...... phosphorylated by purified casein kinase II and of phosphorylated nucleolin which had been isolated from tumor cells grown in the presence of [32P]-o-phosphate, were identical. The partial tryptic digest revealed nine phosphopeptides. Nucleolin isolated from Krebs II mouse ascites cells was phosphorylated by...... purified casein kinase II with about two moles phosphate per one mole of nucleolin....

  15. αS1-casein, which is essential for efficient ER-to-Golgi casein transport, is also present in a tightly membrane-associated form

    Directory of Open Access Journals (Sweden)

    Le Parc Annabelle

    2010-08-01

    Full Text Available Abstract Background Caseins, the main milk proteins, aggregate in the secretory pathway of mammary epithelial cells into large supramolecular structures, casein micelles. The role of individual caseins in this process and the mesostructure of the casein micelle are poorly known. Results In this study, we investigate primary steps of casein micelle formation in rough endoplasmic reticulum-derived vesicles prepared from rat or goat mammary tissues. The majority of both αS1- and β-casein which are cysteine-containing casein was dimeric in the endoplasmic reticulum. Saponin permeabilisation of microsomal membranes in physico-chemical conditions believed to conserve casein interactions demonstrated that rat immature β-casein is weakly aggregated in the endoplasmic reticulum. In striking contrast, a large proportion of immature αS1-casein was recovered in permeabilised microsomes when incubated in conservative conditions. Furthermore, a substantial amount of αS1-casein remained associated with microsomal or post-ER membranes after saponin permeabilisation in non-conservative conditions or carbonate extraction at pH11, all in the presence of DTT. Finally, we show that protein dimerisation via disulfide bond is involved in the interaction of αS1-casein with membranes. Conclusions These experiments reveal for the first time the existence of a membrane-associated form of αS1-casein in the endoplasmic reticulum and in more distal compartments of the secretory pathway of mammary epithelial cells. Our data suggest that αS1-casein, which is required for efficient export of the other caseins from the endoplasmic reticulum, plays a key role in early steps of casein micelle biogenesis and casein transport in the secretory pathway.

  16. Phosphorylation of varicella-zoster virus glycoprotein gpI by mammalian casein kinase II and casein kinase I

    International Nuclear Information System (INIS)

    Varicella-zoster virus (VZV) glycoprotein gpI is the predominant viral glycoprotein within the plasma membranes of infected cells. This viral glycoprotein is phosphorylated on its polypeptide backbone during biosynthesis. In this report, the authors investigated the protein kinases which participate in the phosphorylation events. Under in vivo conditions, VZV gpI was phosphorylated on its serine and threonine residues by protein kinases present within lysates of either VZV-infected or uninfected cells. Because this activity was diminished by heparin, a known inhibitor of casein kinase II, isolated gpI was incubated with purified casein kinase II and shown to be phosphorylated in an in vitro assay containing [γ-32P]ATP. The same glycoprotein was phosphorylated when [32P]GTP was substituted for [32P]ATP in the protein kinase assay. They also tested whether VZV gpI was phosphorylated by two other ubiquitous mammalian protein kinases--casein kinase I and cyclic AMP-dependent kinase--and found that only casein kinase I modified gpI. When the predicted 623-amino-acid sequence of gpI was examined, two phosphorylation sites known to be optimal for casein kinase II were observed. In summary, this study showed that VZV gpI was phosphorylated by each of two mammalian protein kinases (casein kinase I and casein kinase II) and that potential serine-threonine phosphorylation sites for each of these two kinases were present in the viral glycoprotein

  17. Immune activation by casein dietary antigens in bipolar disorder

    NARCIS (Netherlands)

    Severance, E.G.; Dupont, D.; Dickerson, F.B.; Stallings, C.R.; Origoni, A.E.; Krivogorsky, B.; Yang, S.; Haasnoot, W.; Yolken, R.H.

    2010-01-01

    Objectives: Inflammation and other immune processes are increasingly linked to psychiatric diseases. Antigenic triggers specific to bipolar disorder are not yet defined. We tested whether antibodies to bovine milk caseins were associated with bipolar disorder, and whether patients recognized differe

  18. Structural characterization of casein micelles: shape changes during film formation

    International Nuclear Information System (INIS)

    The objective of this study was to determine the effect of size-fractionation by centrifugation on the film structure of casein micelles. Fractionated casein micelles in solution were asymmetrically distributed with a small distribution width as measured by dynamic light scattering. Films prepared from the size-fractionated samples showed a smooth surface in optical microscopy images and a homogeneous microstructure in atomic force micrographs. The nano- and microstructure of casein films was probed by micro-beam grazing incidence small angle x-ray scattering (μGISAXS). Compared to the solution measurements, the sizes determined in the film were larger and broadly distributed. The measured GISAXS patterns clearly deviate from those simulated for a sphere and suggest a deformation of the casein micelles in the film. (paper)

  19. Geographic distribution of haplotype diversity at the bovine casein locus

    OpenAIRE

    Jann, Oliver; Ibeagha-Awemu, Eveline; Özbeyaz, Ceyhan; Zaragoza, Pilar; Williams, John; Ajmone-Marsan, Paolo; Lenstra, Johannes; Moazami-Goudarzi, Katy; Erhardt, Georg

    2004-01-01

    The genetic diversity of the casein locus in cattle was studied on the basis of haplotype analysis. Consideration of recently described genetic variants of the casein genes which to date have not been the subject of diversity studies, allowed the identification of new haplotypes. Genotyping of 30 cattle breeds from four continents revealed a geographically associated distribution of haplotypes, mainly defined by frequencies of alleles at CSN1S1 and CSN3. The genetic diversity within taurine b...

  20. Thermodynamic incompatibility and complex formation in pectin/caseinate mixtures

    OpenAIRE

    Rediguieri, Camila F; de Freitas, Osvaldo; Lettinga, Pavlik; Tuinier, Remco

    2007-01-01

    The instability of polysaccharide/protein mixtures occurs because of either thermodynamic incompatibility or complexation. We studied which instability mechanism dominated given the external conditions. Therefore the effect of temperature, pH, and biopolymer concentration on the phase separation of pectin/caseinate mixtures was investigated. At pH > 6, thermodynamic incompatibility with spinodal decomposition was observed in pectin/caseinate mixtures resulting in the formation of water-in-wat...

  1. Examination of rheological properties of aqueous solutions of sodium caseinate

    Directory of Open Access Journals (Sweden)

    Jolanta Gawałek

    2012-12-01

    Full Text Available Application of sodium caseinate as a functional additive in manufacturing processes requires production of its concentrated aqueous solutions which, in industrial conditions, presents a number of difficulties. In order to develop an effective and optimal industrial process of mixing – manufacturing a concentrated solution of sodium caseinate, it is essential to know rheological properties in a definite range of concentrations changing in the course of the dissolving process. The material for investigations was typical commercial sodium caseinate in the form of dry powder manufactured in Poland from acid casein using the method of extrusion. The objective of the undertaken empirical studies was the assessment of the impact of the concentration on rheological properties of sodium caseinate concentrates. Investigations were carried out for five concentrates manufactured in a mixer equipped in a mechanical agitator at concentrations ranging X (% Î (2.5¸12.5 and changing mass proportions of sodium caseinate in the aqueous solution as follows: GS/G (kgS·kg-1 = 0.025. On the basis of the obtained research results, classical flow curves were plotted for individual concentrates. The determined values of viscosity and density of the examined solutions were correlated in the form of h = f(GS/G and r = f(GS/G dependencies which were used during the determination of classical characteristics of mixing forces essential for the assessment of energetic expenditures required to manufacture concentrates in a mixer equipped in a mechanical agitator. The density of the examined concentrates increased in a way directly proportional, while the dynamic viscosity coefficient increased exponentially together with the increase of sodium caseinate concentration. Sodium caseinate concentrates exhibited Newtonian character in the examined range of concentrations.

  2. Geographic distribution of haplotype diversity at the bovine casein locus.

    OpenAIRE

    Moazami-Goudarzi Katy; Lenstra Johannes A; Ajmone-Marsan Paolo; Williams John L; Zaragoza Pilar; Özbeyaz Ceyhan; Ibeagha-Awemu Eveline M; Jann Oliver C; Erhardt Georg

    2004-01-01

    Abstract The genetic diversity of the casein locus in cattle was studied on the basis of haplotype analysis. Consideration of recently described genetic variants of the casein genes which to date have not been the subject of diversity studies, allowed the identification of new haplotypes. Genotyping of 30 cattle breeds from four continents revealed a geographically associated distribution of haplotypes, mainly defined by frequencies of alleles at CSN1S1 and CSN3. The genetic diversity within ...

  3. Interactions in micellar solutions of β-casein

    Science.gov (United States)

    Leclerc, E.; Calmettes, P.

    1997-02-01

    β-casein is a protein which forms micelles in aqueous solvents. The magnitude and the range of the weight-average interactions between the diverse solute particles are infrared from small-angle neutron scattering measurements made on various β-casein solutions. Well above the critical micelle concentration (CMC), these interactions are repulsive. They weaken with decreasing protein concentration, and finally become strongly attractive near the CMC. Although indispensable for micelle formation this fact has never been reported so far.

  4. Casein Micelles: Size Distribution in Milks from Individual Cows

    OpenAIRE

    Kruif, C.G. de; Huppertz, T.

    2012-01-01

    The size distribution and protein composition of casein micelles in the milk of Holstein-Friesian cows was determined as a function of stage and number of lactations. Protein composition did not vary significantly between the milks of different cows or as a function of lactation stage. Differences in the size and polydispersity of the casein micelles were observed between the milks of different cows, but not as a function of stage of milking or stage of lactation and not even over successive ...

  5. Origin of suppressed demixing in casein/xanthan mixtures

    OpenAIRE

    van Gruijthuijsen, K.; Herle, V.; Tuinier, R.; Schurtenberger, P; Stradner, A.

    2012-01-01

    We explore the properties of casein/xanthan mixtures for xanthan concentrations beyond those inducing phase separation. Previous work has successfully described the onset of demixing by depletion theory in the protein limit, where the xanthan polysaccharides , the polymers , are larger than the caseins from skim milk powder, the colloids (S. Bhat et al., J. Phys.: Condens. Matter, 2006, L339). We now extend these studies to xanthan concentrations in a range of c/c* = 13–88, aiming to arrest t...

  6. Na-caseinate/oil/water systems: emulsion morphology diagrams.

    Science.gov (United States)

    Tan, Hui Lin; McGrath, Kathryn M

    2012-09-01

    The concentrated (dispersed phase 50-70 wt%) composition space of Na-caseinate, a family of milk proteins, stabilised emulsions was investigated for three different oils: soybean oil, palm olein and tetradecane with pH 6.8 phosphate buffer continuous phase. The variation of emulsion stability and microstructure were explored using static light scattering, diffusion nuclear magnetic resonance, cryo-scanning electron microscopy, rheology and the time varying macroscopic phase separation of the emulsions. For soybean oil and palm olein a rich diversity of emulsion microstructures and stabilities are realised. Five emulsion domains, each having a different microstructure and macroscopic stability have been identified within the composition space probed. For the lowest concentrations of emulsifier bridging flocculation is evident and emulsions are of low stability. Increasing Na-caseinate concentration leads to an increased stability and the existence of distinct individual oil droplets, visualised using cryo-scanning electron microscopy. Further increases in Na-caseinate concentration reduce emulsion stability due to depletion flocculation. Na-caseinate self-assembly is then initiated. At sufficiently high Na-caseinate and/or oil concentrations the continuous phase of the emulsion is a three-dimensional protein network and emulsion stability is again enhanced. At the limits of the emulsion composition space a gel-like paste is formed. The diversity of emulsion microstructure is reduced when tetradecane is the discrete phase. Na-caseinate self-assembly is limited and there is no evidence for formation of a protein network. PMID:22709624

  7. Cloning and sequencing of the gene for human. beta. -casein

    Energy Technology Data Exchange (ETDEWEB)

    Loennerdal, B.; Bergstroem, S.; Andersson, Y.; Hialmarsson, K.; Sundgyist, A.; Hernell, O. (Univ. of California, Davis (United States))

    1990-02-26

    Human {beta}-casein is a major protein in human milk. This protein is part of the casein micelle and has been suggested to have several physiological functions in the newborn. Since there is limited information on {beta}casein and the factors that affect its concentration in human milk, the authors have isolated and sequenced the gene for this protein. A human mammary gland cDNA library (Clontech) in gt 11 was screened by plaque hy-hybridization using a 42-mer synthetic {sup 32}p-labelled oligo-nucleotide. Positive clones were identified and isolated, DNA was prepared and the gene isolated by cleavage with EcoR1. Following subcloning (PUC18), restriction mapping and Southern blotting, DNA for sequencing was prepared. The gene was sequenced by the dideoxy method. Human {beta}-casein has 212 amino acids and the amino acid sequence deducted from the nucleotide sequence is to 91% identical to the published sequence for human {beta}-casein show a high degree of conservation at the leader peptide and the highly phosphorylated sequences, but also deletions and divergence at several positions. These results provide insight into the structure of the human {beta}-casein gene and will facilitate studies on factors affecting its expression.

  8. Single-Strand Conformation Polymorphism (SSCP) analysis of alfas1-casein, beta-casein and k-casein genes in Charnequeira Portuguese indigenous goat breed

    OpenAIRE

    Bonifácio, C.; Santos, Ingrid; Belo, Carmona; Cravador, A.

    2001-01-01

    The DNA from eighty goats belonging to the indigenous portuguese caprine breed Charnequeira, ecotype from Beira Baixa (Beiroa), was analysed. Single-strand conformation polymorphisms were identified at exon 9 and at exons 10-11 of the as1-casein gene and at exon 4 of the kcasein. The b-casein gene was found monomorphic at exon 7 in this population sample. The establishment of an association of some of these SSCP polymorphisms with milk production and protein and fat content was at...

  9. Mesoscale crystallization of calcium phosphate nanostructures in protein (casein) micelles

    Science.gov (United States)

    Thachepan, Surachai; Li, Mei; Mann, Stephen

    2010-11-01

    Aqueous micelles of the multi-protein calcium phosphate complex, casein, were treated at 60 °C and pH 7 over several months. Although partial dissociation of the micelles into 12 nm sized amorphous calcium phosphate (ACP)/protein nanoparticles occurred within a period of 14 days, crystallization of the ACP nanoclusters into bundles of hydroxyapatite (HAP) nanofilaments was not observed until after 12 weeks. The HAP nanofilaments were formed specifically within the partially disrupted protein micelles suggesting a micelle-mediated pathway of mesoscale crystallization. Similar experiments using ACP-containing synthetic micelles prepared from β-casein protein alone indicated that co-aligned bundles of HAP nanofilaments were produced within the protein micelle interior after 24 hours at temperatures as low as 35 °C. The presence of Mg2+ ions in the casein micelles, as well as a possible synergistic effect associated with the multi-protein nature of the native aggregates, could account for the marked inhibition in mesoscale crystallization observed in the casein micelles compared with the single-component β-casein constructs.Aqueous micelles of the multi-protein calcium phosphate complex, casein, were treated at 60 °C and pH 7 over several months. Although partial dissociation of the micelles into 12 nm sized amorphous calcium phosphate (ACP)/protein nanoparticles occurred within a period of 14 days, crystallization of the ACP nanoclusters into bundles of hydroxyapatite (HAP) nanofilaments was not observed until after 12 weeks. The HAP nanofilaments were formed specifically within the partially disrupted protein micelles suggesting a micelle-mediated pathway of mesoscale crystallization. Similar experiments using ACP-containing synthetic micelles prepared from β-casein protein alone indicated that co-aligned bundles of HAP nanofilaments were produced within the protein micelle interior after 24 hours at temperatures as low as 35 °C. The presence of Mg2+ ions in

  10. Casein maps: Effect of ethanol, pH, temperature, and CaCl2 on the particle size of reconstituted casein micelles

    OpenAIRE

    Ye, Ran; Harte, Federico

    2012-01-01

    Although conditions favoring casein micelle aggregation are well known, factors promoting the dissociation of the casein micelle are not fully understood. It was our objective to investigate the ethanol-induced dissociation of micellar casein as affected by temperature and a wide range of pH, along with the concentrations of calcium and casein. Two different concentrations of casein micelles were dispersed in imidazole buffer with 0 to 80% ethanol (vol/vol) and 2 and 10 mM calcium. Apparent m...

  11. αS1-casein, which is essential for efficient ER-to-Golgi casein transport, is also present in a tightly membrane-associated form

    OpenAIRE

    Le Parc Annabelle; Leonil Joëlle; Chanat Eric

    2010-01-01

    Abstract Background Caseins, the main milk proteins, aggregate in the secretory pathway of mammary epithelial cells into large supramolecular structures, casein micelles. The role of individual caseins in this process and the mesostructure of the casein micelle are poorly known. Results In this study, we investigate primary steps of casein micelle formation in rough endoplasmic reticulum-derived vesicles prepared from rat or goat mammary tissues. The majority of both αS1- and β-casein which a...

  12. Structural investigations of sodium caseinate micelles in complex environments

    International Nuclear Information System (INIS)

    Full text: The most frequent destabilization mechanisms in Sodium Caseinate (NaCas) emulsions are creaming and flocculation. Coarse or fine emulsions with low protein con- tent destabilize mainly by creaming. If migration mechanism is suppressed, flocculation may become the main mechanism of destabilization. Small Angle X-Ray Scattering (SAXS) technique was applied to investigate sodium caseinate micelles structure in different environments. As many natural products, Sodium Caseinate samples have large polydisperse size distribution. The experimental data was analyzed using advanced modeling approaches. The Form Factor for the Caseinate micelle subunits was described by an ellipsoidal core shell model and the structure factor was split into two contributions, one corresponding to the particle-particle interactions and another one for the long range correlation of the subunits in the supramolecular structure. For the first term the hard sphere structure factor using the Percus-Yevick approximation for closure relation was used and for the second term a fractal model was applied. Three concentrations of sodium Caseinate (2, 5 and 7.5 %wt.) were measured in pure water, sugar solutions (20 %wt.) and in three different lipid phase emulsions containing 10 %wt. sunflower seed, olive and fish oils. Data analysis provided an average casein subunit radius of 4 nm, an average distance between the subunits of around 20nm and a fractal dimension value of around 3 for all samples. As indicated by the values of the correlation lengths for the set of studied samples, the casein aggregation is strongly affected by simple sugar additions and it is enhanced by emulsion droplets hydrophobic interaction. As will be presented, these nanoscale structural results provided by scattering experiments is consistent with macroscopic results obtained from several techniques, providing a new understanding of NaCas emulsions. (author)

  13. Structural investigations of sodium caseinate micelles in complex environments

    Energy Technology Data Exchange (ETDEWEB)

    Huck Iriart, C.; Herrera, M.L.; Candal, R. [Universidad de Buenos Aires, Buenos Aires (Argentina); Oliveira, C.L.P. [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil); Torriani, I. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil)

    2012-07-01

    Full text: The most frequent destabilization mechanisms in Sodium Caseinate (NaCas) emulsions are creaming and flocculation. Coarse or fine emulsions with low protein con- tent destabilize mainly by creaming. If migration mechanism is suppressed, flocculation may become the main mechanism of destabilization. Small Angle X-Ray Scattering (SAXS) technique was applied to investigate sodium caseinate micelles structure in different environments. As many natural products, Sodium Caseinate samples have large polydisperse size distribution. The experimental data was analyzed using advanced modeling approaches. The Form Factor for the Caseinate micelle subunits was described by an ellipsoidal core shell model and the structure factor was split into two contributions, one corresponding to the particle-particle interactions and another one for the long range correlation of the subunits in the supramolecular structure. For the first term the hard sphere structure factor using the Percus-Yevick approximation for closure relation was used and for the second term a fractal model was applied. Three concentrations of sodium Caseinate (2, 5 and 7.5 %wt.) were measured in pure water, sugar solutions (20 %wt.) and in three different lipid phase emulsions containing 10 %wt. sunflower seed, olive and fish oils. Data analysis provided an average casein subunit radius of 4 nm, an average distance between the subunits of around 20nm and a fractal dimension value of around 3 for all samples. As indicated by the values of the correlation lengths for the set of studied samples, the casein aggregation is strongly affected by simple sugar additions and it is enhanced by emulsion droplets hydrophobic interaction. As will be presented, these nanoscale structural results provided by scattering experiments is consistent with macroscopic results obtained from several techniques, providing a new understanding of NaCas emulsions. (author)

  14. The role of casein in supporting the operation of surface bound kinesin

    Directory of Open Access Journals (Sweden)

    Hancock William O

    2008-10-01

    Full Text Available Abstract Microtubules and associated motor proteins such as kinesin are envisioned for applications such as bioseparation and molecular sorting to powering hybrid synthetic mechanical devices. One of the challenges in realizing such systems is retaining motor functionality on device surfaces. Kinesin motors adsorbed onto glass surfaces lose their functionality or ability to interact with microtubules if not adsorbed with other supporting proteins. Casein, a milk protein, is commonly used in microtubule motility assays to preserve kinesin functionality. However, the mechanism responsible for this preservation of motor function is unknown. To study casein and kinesin interaction, a series of microtubule motility assays were performed where whole milk casein, or its αs1 and αs2, β or κ subunits, were introduced or omitted at various steps of the motility assay. In addition, a series of epifluorescence and total internal reflection microscopy (TIRF experiments were conducted where fluorescently labeled casein was introduced at various steps of the motility assay to assess casein-casein and casein-glass binding dynamics. From these experiments it is concluded that casein forms a bi-layer which supports the operation of kinesin. The first tightly bound layer of casein mainly performs the function of anchoring the kinesin while the second more loosely bound layer of casein positions the head domain of the kinesin to more optimally interact with microtubules. Studies on individual casein subunits indicate that β casein was most effective in supporting kinesin functionality while κ casein was found to be least effective.

  15. The decontamination of industrial casein and milk powder by irradiation

    Science.gov (United States)

    Żegota, H.; Małolepszy, B.

    2008-09-01

    The efficacy of gamma radiation decontamination of industrial casein, a milk protein utilized as a component of many food and non-food products has been studied. Low-fat milk powder was also included with a purpose to study the microflora survival in protein-rich materials. Microbial analysis of the samples prior to irradiation showed that the initial total viable count was higher than 6.0 log cfu g -1 in both casein and milk powders. The contamination of casein with moulds and yeasts was found to be equal to 3.56 log cfu g -1. The counts of coliforms have not exceeded the value of 2.48 log cfu g -1. Radiation processing of casein and milk powder has substantially reduced the microbial population of all samples. The dose of 5 kGy was sufficient to reduce the total microflora and coliforms counts to the level permitted for food products. Survivals of microorganisms were analyzed by the generalized exponential equation, SF =exp[ -D/ Do) α]. Values of an exponent, α, standing for the dispersion parameter, were equal to 0.65 and 0.70 for microorganisms contaminating casein and milk powders, respectively. The numerical value of the dispersion parameter α<1 indicates the concave dependence of a logarithm of surviving fraction versus radiation dose. No difference in microflora survival in irradiated samples tested immediately and in samples stored for 1-month after irradiation has been noticed.

  16. Human alpha s1-casein: purification and characterization.

    Science.gov (United States)

    Rasmussen, L K; Due, H A; Petersen, T E

    1995-05-01

    The human counterpart of alpha s1-casein has been purified by a combination of gel-filtration and ion-exchange chromatography under denaturing conditions. SDS-PAGE analysis revealed the presence of a diffuse ladder with a high molecular mass which upon reduction was replaced by several closely spaced bands of lower molecular masses and a broad diffuse band corresponding to kappa-casein. Amino acid sequence analysis of the closely spaced bands all resulted in the same N-terminal sequence which was found to be homologous with alpha s1-casein from other species. Sequence analysis of a major radiolabelled tryptic peptide from purified 14C-carboxymethylated alpha s1-casein demonstrated that the protein contains at least two cysteine residues. As judged by SDS-PAGE in the presence or absence of a reducing agent, the molecular structure of the polymers constituting the ladder is composed of heteropolymers of alpha s1- and kappa-casein cross-linked by disulfide bonds. PMID:7749638

  17. The decontamination of industrial casein and milk powder by irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Zegota, H. [Institute of Applied Radiation Chemistry, Technical University, Wroblewskiego 15, 93-590 Lodz (Poland)], E-mail: ahzegota@mitr.p.lodz.pl; Malolepszy, B. [Fleur Comp. Ltd., Artyleryjska 6, 91-072 Lodz (Poland)

    2008-09-15

    The efficacy of gamma radiation decontamination of industrial casein, a milk protein utilized as a component of many food and non-food products has been studied. Low-fat milk powder was also included with a purpose to study the microflora survival in protein-rich materials. Microbial analysis of the samples prior to irradiation showed that the initial total viable count was higher than 6.0 log cfu g{sup -1} in both casein and milk powders. The contamination of casein with moulds and yeasts was found to be equal to 3.56 log cfu g{sup -1}. The counts of coliforms have not exceeded the value of 2.48 log cfu g{sup -1}. Radiation processing of casein and milk powder has substantially reduced the microbial population of all samples. The dose of 5 kGy was sufficient to reduce the total microflora and coliforms counts to the level permitted for food products. Survivals of microorganisms were analyzed by the generalized exponential equation, SF=exp[-D/D{sub o}){sup {alpha}}]. Values of an exponent, {alpha}, standing for the dispersion parameter, were equal to 0.65 and 0.70 for microorganisms contaminating casein and milk powders, respectively. The numerical value of the dispersion parameter {alpha}<1 indicates the concave dependence of a logarithm of surviving fraction versus radiation dose. No difference in microflora survival in irradiated samples tested immediately and in samples stored for 1-month after irradiation has been noticed.

  18. Cryo-transmission electron tomography of native casein micelles from bovine milk

    OpenAIRE

    Trejo, R.; Dokland, T; Jurat-Fuentes, J.; Harte, F.

    2011-01-01

    Caseins are the principal protein components in milk and an important ingredient in the food industry. In liquid milk, caseins are found as micelles of casein proteins and colloidal calcium nanoclusters. Casein micelles were isolated from raw skim milk by size exclusion chromatography and suspended in milk protein-free serum produced by ultrafiltration (molecular weight cut-off of 3 kDa) of raw skim milk. The micelles were imaged by cryo-electron microscopy and subjected to tomographic recons...

  19. Evolution of the casein multigene family: conserved sequences in the 5' flanking and exon regions.

    OpenAIRE

    Yu-Lee, L Y; Richter-Mann, L; Couch, C H; Stewart, A F; Mackinlay, A G; Rosen, J. M.

    1986-01-01

    The rat alpha- and bovine alpha s1-casein genes have been isolated and their 5' sequences determined. The rat alpha-, beta-, gamma- and bovine alpha s1-casein genes contain similar 5' exon arrangements in which the 5' noncoding, signal peptide and casein kinase phosphorylation sequences are each encoded by separate exons. These findings support the hypothesis that during evolution, the family of casein genes arose by a process involving exon recruitment followed by intragenic and intergenic d...

  20. Kappa Casein Gene Polymorphism in Holstein Chinese Cattle

    Directory of Open Access Journals (Sweden)

    A. E. Hamza1,*, X. L. Wang and Z. P.Yang

    2010-10-01

    Full Text Available Kappa casein gene polymorphism has received a considerable attention because of its correlation with milk quality, composition and technological properties. The polymorphism of kappa casein gene (K-CN was detected in Holstein Chinese cattle. A 218 bp sequence in exon IV of 319 Holstein Chinese cattle blood samples were amplified using polymerase chain reaction-single strand conformation (PCR-SSCP technique. Sequence analysis revealed one single nucleotide polymorphism (SNP T/C SNP in exon 1V at nucleotides (80, moreover; three genotypes TT, TC and CC were also identified with following frequencies: 0.40, 0.34 and 0.26%, respectively. The allele frequency for T and C found to be 0.6 and 0.4 %, respectively. Allele frequencies in the population fitted with Hardy–Weinberg equilibrium (P>0.05. Analysis of genetic polymorphism of k-casein at exon 1V exhibited medium polymorphism information content (PIC=0.36.

  1. Hollow Casein-Based Polymeric Nanospheres for Opaque Coatings.

    Science.gov (United States)

    Zhang, Fan; Ma, Jianzhong; Xu, Qunna; Zhou, Jianhua; Simion, Demetra; Carmen, Gaidău; Wang, John; Li, Yunqi

    2016-05-11

    Casein-based hollow polymeric sphere were fabricated through emulsifier-free polymerization coupled with alkali swelling approach. Hollow structure and nanoscale size of casein-based polymeric spheres were verified by TEM, AFM, SEM, and UV-vis spectra. The as-obtained hollow spheres were proved exhibiting superior opaque characteristic. Through adjusting the structural parameters, for example, MAA usages and MAA content in seed to core, sphere film showed tunable visible-light transmittance and antiultraviolet property. The formation mechanism of casein-based hollow sphere has been discussed in depth. Worth mentioning, the resultant hollow polymeric sphere can easily form films itself at room temperature, which would open a new possibility of designing opaque coatings in several fields, such as leather, packaging, paper making, biomedical, and special indoor coating applications. PMID:27090208

  2. Rheology and phase behavior of dense casein micelle dispersions

    Science.gov (United States)

    Bouchoux, A.; Debbou, B.; Gésan-Guiziou, G.; Famelart, M.-H.; Doublier, J.-L.; Cabane, B.

    2009-10-01

    Casein micelle dispersions have been concentrated through osmotic stress and examined through rheological experiments. In conditions where the casein micelles are separated from each other, i.e., below random-close packing, the dispersions have exactly the flow and dynamic properties of the polydisperse hard-sphere fluid, demonstrating that the micelles interact only through excluded volume effects in this regime. These interactions cause the viscosity and the elastic modulus to increase by three orders of magnitude approaching the concentration of random-close packing estimated at Cmax≈178 g/l. Above Cmax, the dispersions progressively turn into "gels" (i.e., soft solids) as C increases, with elastic moduli G' that are nearly frequency independent. In this second regime, the micelles deform and/or deswell as C increases, and the resistance to deformation results from the formation of bonds between micelles combined with the intrinsic mechanical resistance of the micelles. The variation in G' with C is then very similar to that observed with concentrated emulsions where the resistance to deformation originates from a set of membranes that separate the droplets. As in the case of emulsions, the G' values at high frequency are also nearly identical to the osmotic pressures required to compress the casein dispersions. The rheology of sodium caseinate dispersions in which the caseins are not structured into micelles is also reported. Such dispersions have the behavior of associative polymer solutions at all the concentrations investigated, further confirming the importance of structure in determining the rheological properties of casein micelle systems.

  3. Molecular modeling and conformational IgG epitope mapping on bovine β-casein

    NARCIS (Netherlands)

    Liu, Fahui; Gao, Jinyan; Li, Xin; Chen, Hongbing

    2016-01-01

    Characterizing conformational B-cell epitopes is a key step for understanding the immunological basis of allergy contributed by β-casein. There is no resolved conformational structure of β-casein in protein data bank, and most of the previous research on epitope identification of β-casein focused

  4. Electrochemical immunosensor for the determination of β-casein

    Directory of Open Access Journals (Sweden)

    Judith Molinari

    2015-03-01

    Full Text Available An amperometric biosensor for the quantification of food allergens based on an inhibitory immunoassay is presented. As a proof of concept, the experimental conditions were optimized for the detection of β-casein in the 0-10 ppm range. Eight electro­che­mi­cal cells were integrated into a small-sized portable potentiostat controlled by a smart­phone via Bluetooth communication. The determination of β-casein in eight different samples can be measured with the electrochemical biosensor, which has the potential to be modified for the detection of multiple allergens.

  5. Effects of casein and fat content on water self-diffusion coefficients in casein systems: a pulsed field gradient nuclear magnetic resonance study

    OpenAIRE

    A. Metais; Cambert, M.; Riaublanc, A.; Mariette, F.

    2004-01-01

    The water self-diffusion coefficients in casein matrixes were measured using a pulsed field gradient spin-echo nuclear magnetic resonance technique (PFG-SE NMR). The dependence of the water self-diffusion coefficient on the casein concentration and the aqueous phase composition is reported in both a rehydrated native phosphocaseinate dispersion and a concentrated casein retentate. A model has been proposed to explain the different behavior of the water self-diffusion coefficient in the two ca...

  6. Biochemical and functional characterisation of casein and whey protein hydrolysates.

    OpenAIRE

    Ven, van de, P.M.

    2002-01-01

    Whey protein and sodium caseinate were hydrolysed with commercially available enzyme preparations. The resulting hydrolysates were characterised using several analytical characterisation methods and by determination of several functional properties. Subsequently, correlations between the biochemical characteristics themselves and between biochemical and functional properties were studied using multivariate regression analysis.Biochemical characteristics of hydrolysates were determined using u...

  7. Process standardization for rennet casein based Mozzarella cheese analogue.

    Science.gov (United States)

    Shah, Rahul; Jana, Atanu H; Aparnathi, K D; Prajapati, P S

    2010-10-01

    A process for manufacture of Mozzarella cheese analogue (MCA) using rennet casein and plastic cream as protein and fat sources respectively was standardized. The formulation comprised of 25% plastic cream (72% fat), 27% rennet casein along with 3% tri-sodium citrate as emulsifying salt, 2% maltodextrin as binder, 0.55% lactic acid as pH regulator, 1% common salt for seasoning, 1% Mozzarella cheese bud as flavouring and 40.4% water. The process involved (a) dissolving the dry mixture of casein, maltodextrin, flavouring and common salt in hot emulsifying salt solution, (b) incorporation of half the quantity of acid solution in casein-maltodextrin dough, followed by addition and emulsification of plastic cream, and (c) addition of remaining half of the acid solution and heating the mass to 80 °C until a plastic cheese mass was obtained. The analogue was shaped in ball form, cooled and packaged in polyethylene bag. The MCA conformed to the PFA requirements for pizza cheese and had all the requisite baking characteristics expected of pizza cheese topping. PMID:23572688

  8. CALCIUM-INDUCED SUPRAMOLECULAR STRUCTURES IN THE CALCIUM CASEINATE SYSTEM

    Science.gov (United States)

    The molecular details deciphering the spontaneous calcium-induced protein aggregation process in the calcium caseinate system remain obscure. Understanding this complex process could lead to potential new applications of this important food ingredient. In this work, we studied calcium-induced supra...

  9. CASEIN MICELLE STRUCTURE: THE PAST AND THE PRESENT

    Science.gov (United States)

    At the heart of the milk system are the colloidal casein–calcium–transport complexes termed the casein micelles. The application of physical chemical techniques such as light, neutron, and X-ray scattering, and Electron Microscopy (EM) has yielded a wealth of experimental detail concerning the struc...

  10. The physical characteristics and emulsification properties of partially dephosphorylated bovine β-casein.

    Science.gov (United States)

    McCarthy, Noel A; Kelly, Alan L; O'Mahony, James A; Fenelon, Mark A

    2013-06-01

    Bovine β-casein was purified from phosphocasein by rennet coagulation and cold solubilisation from the resultant curd. β-Casein was then dephosphorylated using potato acid phosphatase. Urea-polyacrylamide gel electrophoresis (PAGE) of partially dephosphorylated β-casein showed a number of bands, depending on the final level of phosphorylation. Dephosphorylating β-casein increased its pH of minimum solubility from ∼pH 5 to 5.5 and reduced its net negative charge from -30.8 to -27.0 mV. During the acidification of β-casein solutions, partially dephosphorylated β-casein failed to form a gel, unlike the phosphorylated (i.e., control) β-casein. Use of partially dephosphorylated β-casein to stabilise oil-in-water emulsions resulted in larger fat globules compared to control β-casein, but such globules were less susceptible to aggregation in the presence of 15 or 30 mM CaCl(2). Overall, the dephosphorylation of β-casein resulted in a protein similar to human β-casein in terms of physicochemical functionality, with increased stability against calcium-induced aggregation. PMID:23411247

  11. Order of 17 july 1991 on treatment by ionizing radiation of casein and casein products for human consumption

    International Nuclear Information System (INIS)

    This Order authorizes and fixes the conditions for the sale and marketing of casein (one of the chief constituents of milk which forms the basis of cheese), which is treated by ionizing radiation, for human consumption. The absorbed radiation dose must not exceed 6 kGy

  12. PFG-NMR self-diffusion in casein dispersions: Effects of probe size and protein aggregate size

    OpenAIRE

    Salami, S; Rondeau Mouro, C.; Van Duyhoven, J.; Mariette, F.

    2013-01-01

    The self-diffusion coefficients of different molecular weight PEGs (Polyethylene glycol) and casein particles were measured, using a pulsed-gradient nuclear magnetic resonance technique (PFG-NMR), in native phosphocaseinate (NPC) and sodium caseinate (SC) dispersions where caseins are not structured into micelles. The dependence of the PEG self-diffusion coefficient on the PEG size, casein concentration, the size and the mobility of casein obstacle particles are reported. Wide differences in ...

  13. Milk lacking α-casein leads to permanent reduction in body size in mice.

    Directory of Open Access Journals (Sweden)

    Andreas F Kolb

    Full Text Available The major physiological function of milk is the transport of amino acids, carbohydrates, lipids and minerals to mammalian offspring. Caseins, the major milk proteins, are secreted in the form of a micelle consisting of protein and calcium-phosphate.We have analysed the role of the milk protein α-casein by inactivating the corresponding gene in mice. Absence of α-casein protein significantly curtails secretion of other milk proteins and calcium-phosphate, suggesting a role for α-casein in the establishment of casein micelles. In contrast, secretion of albumin, which is not synthesized in the mammary epithelium, into milk is not reduced. The absence of α-casein also significantly inhibits transcription of the other casein genes. α-Casein deficiency severely delays pup growth during lactation and results in a life-long body size reduction compared to control animals, but has only transient effects on physical and behavioural development of the pups. The data support a critical role for α-casein in casein micelle assembly. The results also confirm lactation as a critical window of metabolic programming and suggest milk protein concentration as a decisive factor in determining adult body weight.

  14. The bioactive effects of casein proteins on enteroendocrine cell health, proliferation and incretin hormone secretion.

    Science.gov (United States)

    Gillespie, Anna L; Green, Brian D

    2016-11-15

    Previous studies suggest that casein exerts various anti-diabetic effects. However, it is not known which casein proteins are bioactive, nor their effects on enteroendocrine cells. This study evaluated the effects of intact whole casein, intact individual proteins (alpha, beta and kappa casein) and hydrolysates on an enteroendocrine cell line. High content analysis accurately monitored changes in cell health and intracellular glucagon-like peptide-1 (GLP-1) content. Cheese ripening duration and GLP-1 secretory responses were also considered. Beta casein significantly stimulated enteroendocrine cell proliferation and all caseins were potent GLP-1 secretagogues (except kappa casein). Interestingly the GLP-1 secretory activity was almost always lost or significantly reduced upon hydrolysis with proteolytic enzymes. Only pepsin-derived beta casein hydrolysates had significantly increased potency compared with the intact protein, but this was diminished with prolonged hydrolysis. In conclusion casein proteins are not detrimental to enteroendocrine cells, and alpha and beta casein are particularly beneficial stimulating proliferation and GLP-1 secretion. PMID:27283618

  15. Influence of race and crossbreeding on casein micelles size.

    Science.gov (United States)

    Freitas, Denise R; Fonseca, Leorges M; Souza, Fernando N; Ladeira, Cristiane V G; Diniz, Soraia A; Haddad, João Paulo A; Ferreira, Diêgo S; Santoro, Marcelo M; Cerqueira, Mônica M O P

    2015-05-01

    Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production. PMID:25488503

  16. Characterization of casein hydrolysates derived from enzymatic hydrolysis

    OpenAIRE

    Wang, Jinshui; Su, Yinjie; Jia, Feng; Jin, Huali

    2013-01-01

    Background Casein is the main proteinaceous component of milk and has made us interest due to its wide applications in the food, drug, and cosmetic industries as well as to its importance as an investigation material for elucidating essential questions regarding the protein chemistry. Enzymatic hydrolysis is an important method commonly used in the modification of protein structure in order to enhance the functional properties of proteins. The relationship between enzymatic hydrolysis and str...

  17. The Size Distribution of Casein Micelles in Camel Milk

    OpenAIRE

    Farah, Z.; Ruegg, M. W.

    1989-01-01

    The size distribution of casein micelles in camel milk has been determined by electron microscopy. Individual and pooled samples were cryo-fixed by rapid freezing and freeze-fractured. Electron micrographs of the freeze-fracture replica revealed a relatively broad size distribution, with an average micelle dimeter around 280 nm in the volume distribution curve. The distribution was significantly broader than that of the particles of cow's or human milk and showed a greater number of large ...

  18. The Size Distribution of Bovine Casein Micelles: A Review

    OpenAIRE

    Holt, C.

    1985-01-01

    This review considers the average size and size distribution of bovine casein micelles as measured by electron microscopy, light scattering and controlled pore glass chromatography, and the origin and biological function of the size distribution. Recent work by electron microscopy has given average sizes in reasonable agreement with measurements on the same milk sample by light scattering . It is suggested that natural variations in averaqe micelle size and overestimation of micelle radii ...

  19. Solving the mystery of the internal structure of casein micelles.

    Science.gov (United States)

    Ingham, B; Erlangga, G D; Smialowska, A; Kirby, N M; Wang, C; Matia-Merino, L; Haverkamp, R G; Carr, A J

    2015-04-14

    The interpretation of milk X-ray and neutron scattering data in relation to the internal structure of the casein micelle is an ongoing debate. We performed resonant X-ray scattering measurements on liquid milk and conclusively identified key scattering features, namely those corresponding to the size of and the distance between colloidal calcium phosphate particles. An X-ray scattering feature commonly assigned to the particle size is instead due to protein inhomogeneities. PMID:25711160

  20. Molecular cloning of the human casein kinase II α subunit

    International Nuclear Information System (INIS)

    A human cDNA encoding the α subunit of casein kinase II and a partial cDNA encoding the rat homologue were isolated by using a Drosophila casein kinase II cDNA probe. The 2.2-kb human cDNA contains a 1.2-kb open reading frame, 150 nucleotides of 5' leader, and 850 nucleotides of 3' noncoding region. Except for the first 7 deduced amino acids that are missing in the rat cDNA, the 328 amino acids beginning with the amino terminus are identical between human and rat. The Drosophila enzyme sequence is 90% identical with the human casein kinase II sequence, and there is only a single amino acid difference between the published partial bovine sequence and the human sequence. In addition, the C-terminus of the human cDNA has an extra 53 amino acids not present in Drosophila. Northern analysis of rat and human RNA showed predominant bands of 5.5, 3.1, and 1.8 kb. In rat tissues, brain and spleen had the highest levels of casein kinase II α subunit specific RNA, while skeletal muscle showed the lowest. Southern analysis of human cultured cell and tissue genomic DNA using the full-length cDNA probe revealed two bands with restriction enzymes that have no recognition sites within the cDNA and three to six bands with enzymes having single internal sites. These results are consistent with the possibility that two genes encode the α subunits

  1. Geographic distribution of haplotype diversity at the bovine casein locus

    Directory of Open Access Journals (Sweden)

    Moazami-Goudarzi Katy

    2004-03-01

    Full Text Available Abstract The genetic diversity of the casein locus in cattle was studied on the basis of haplotype analysis. Consideration of recently described genetic variants of the casein genes which to date have not been the subject of diversity studies, allowed the identification of new haplotypes. Genotyping of 30 cattle breeds from four continents revealed a geographically associated distribution of haplotypes, mainly defined by frequencies of alleles at CSN1S1 and CSN3. The genetic diversity within taurine breeds in Europe was found to decrease significantly from the south to the north and from the east to the west. Such geographic patterns of cattle genetic variation at the casein locus may be a result of the domestication process of modern cattle as well as geographically differentiated natural or artificial selection. The comparison of African Bos taurus and Bos indicus breeds allowed the identification of several Bos indicus specific haplotypes (CSN1S1*C-CSN2*A2-CSN3*AI/CSN3*H that are not found in pure taurine breeds. The occurrence of such haplotypes in southern European breeds also suggests that an introgression of indicine genes into taurine breeds could have contributed to the distribution of the genetic variation observed.

  2. Interactions between tea catechins and casein micelles and their impact on renneting functionality.

    Science.gov (United States)

    Haratifar, Sanaz; Corredig, Milena

    2014-01-15

    Many studies have shown that tea catechins bind to milk proteins. This research focused on the association of tea polyphenols with casein micelles, and the consequences of the interactions on the renneting behaviour of skim milk. It was hypothesized that epigallocatechin-gallate (EGCG), the main catechin present in green tea, forms complexes with the casein micelles and that the association modifies the processing functionality of casein micelles. The binding of EGCG to casein micelles was quantified using HPLC. The formation of catechin-casein micelles complexes affected the rennet induced gelation of milk, and the effect was concentration dependent. Both the primary as well as the secondary stage of gelation were affected. These experiments clearly identify the need for a better understanding of the effect of tea polyphenols on the processing functionality of casein micelles, before milk products can be used as an appropriate platform for delivery of bioactive compounds. PMID:24054208

  3. Scaling relations between structure and rheology of ageing casein particle gels

    OpenAIRE

    Mellema, M

    2000-01-01

    Mellema, M. (Michel), Scaling relations between structure and rheology of ageing casein particle gels , PhD Thesis, Wageningen University, 150 + 10 pages, references by chapter, English and Dutch summaries (2000).The relation between (colloidal) interactions, structure and rheology of particle gels is discussed, especially the properties and the spontaneous ageing behavior of rennet-induced casein(ate) or skim milk gels.Methods involved were Brownian dynamics simulations, confocal microscopy,...

  4. Pressure-induced dissociation of casein micelles: size distribution and effect of temperature

    OpenAIRE

    Gebhardt, R.; Doster, W.; U. Kulozik

    2005-01-01

    Pressure-induced dissociation of a turbid solution of casein micelles was studied in situ in static and dynamic light scattering experiments. We show that at high pressure casein micelles decompose into small fragments comparable in size to casein monomers. At intermediate pressure we observe particles measuring 15 to 20 nm in diameter. The stability against pressure dissociation increased with temperature, suggesting enhanced hydrophobic contacts. The pressure transition curves are biphasic,...

  5. Effect of Calcium Concentration on the Structure of Casein Micelles in Thin Films

    OpenAIRE

    Müller-Buschbaum, P.; Gebhardt, R.; Roth, S. V.; Metwalli, E.; Doster, W.

    2007-01-01

    The structure of thin casein films prepared with spin-coating is investigated as a function of the calcium concentration. Grazing incidence small-angle x-ray scattering and atomic force microscopy are used to probe the micelle structure. For comparison, the corresponding casein solutions are investigated with dynamic light-scattering experiments. In the thin films with added calcium three types of casein structures, aggregates, micelles, and mini-micelles, are observed in coexistence with ato...

  6. Untersuchungen zur Proteolyse von para-k-Casein: vom Modell zum Käse

    OpenAIRE

    Böhm, Anke

    2003-01-01

    Para-k-Casein entsteht durch Hydrolyse des kappa-Caseins nach Zugabe proteolytischer Enzyme zur Milch. Untersuchungen an selbst erstellten Modellen unter Bedingungen, die die Käsereifung simulieren, zeigen, dass die Proteolyse des für die Käsereifung bedeutenden para-k-Caseins stark vom Wassergehalt abhängt. Mit Hilfe geeigneter Methoden (SDS-Elektrophorese, IEF, GPC, RP-HPLC, ESI-MS u.a.) konnte der Abbau des para-k-Caseins durch die industriell relevanten Milchgerinnungsenzyme Chymosin, Fro...

  7. Behaviour of casein micelles at conditions comparable to those in ice cream

    OpenAIRE

    Jonkman, M.J.

    2000-01-01

    The physical properties of ice cream are mainly determined by the processing and the ingredients. Milk (powder) is one of the ingredients and ice cream thus contains casein, the major milk protein. A large proportion of casein in ice cream is present in the plasma phase of ice cream. Since the behaviour of casein in ice cream plasma was not known and could not be predicted and is expected to be important for the properties of ice cream, the behaviour of casein micelles in ice cream plasma and...

  8. Genetic parameters and selection for casein content in Italian Holstein and Brown Swiss

    Directory of Open Access Journals (Sweden)

    Alessandro Bagnato

    2010-01-01

    Full Text Available A total of more than 2,000,000 records on casein contents were collected in Lombardia (Italy during routine milk recording of Italian Holstein and Brown Swiss dairy cows. Variance components for casein were estimated as well as all the genetic correlations of casein with production and type traits considered in selection. According to the heritabilities estimated (12.4% for Brown and 9.36% for Holstein, breeding values were calculated for bulls and compared to the breeding values for total protein. The results of two different selection scenarios were compared for each breed when including protein or casein as selection criterion. Genetic progress expected for all traits selected were compared after 10 years of selection. The genetic variability of casein allows the use of this trait as selection criterion with the estimation of breeding values and its inclusion in selection indexes. Ranking of breeding values for casein and protein are very similar in both breeds. But some differences in genetic values for casein exist for the same level of breeding value for protein. Nevertheless results in genetic gain differ between breeds depending mainly on genetic correlations with the other traits selected. The positive results in selection response estimated for several traits suggest to the Brown Swiss Association the replacement of protein selection with casein. In contrast the smaller effects estimated for the Italian Holstein suggest to wait for more casein data collected before any change in selection program.

  9. Milk Lacking α-Casein Leads to Permanent Reduction in Body Size in Mice

    OpenAIRE

    Kolb, Andreas F.; Huber, Reinhard C.; Simon G. Lillico; Carlisle, Ailsa; Robinson, Claire J.; Neil, Claire; Petrie, Linda; Sorensen, Dorte B.; Olsson, I. Anna S.; Whitelaw, C. Bruce A

    2011-01-01

    The major physiological function of milk is the transport of amino acids, carbohydrates, lipids and minerals to mammalian offspring. Caseins, the major milk proteins, are secreted in the form of a micelle consisting of protein and calcium-phosphate. We have analysed the role of the milk protein α-casein by inactivating the corresponding gene in mice. Absence of α-casein protein significantly curtails secretion of other milk proteins and calcium-phosphate, suggesting a role for α-casein in the...

  10. Characterization of casein and alpha lactalbumin of African elephant (Loxodonta africana) milk.

    Science.gov (United States)

    Madende, M; Osthoff, G; Patterton, H-G; Patterton, H E; Martin, P; Opperman, D J

    2015-12-01

    The current research reports partial characterization of the caseins and α-lactalbumin (α-LA) of the African elephant with proposed unique structure-function properties. Extensive research has been carried out to understand the structure of the casein micelles. Crystallographic structure elucidation of caseins and casein micelles is not possible. Consequently, several models have been developed in an effort to describe the casein micelle, specifically of cow milk. Here we report the characterization of African elephant milk caseins. The κ-caseins and β-caseins were investigated, and their relative ratio was found to be approximately 1:8.5, whereas α-caseins were not detected. The gene sequence of β-casein in the NCBI database was revisited, and a different sequence in the N-terminal region is proposed. Amino acid sequence alignment and hydropathy plots showed that the κ-casein of African elephant milk is similar to that of other mammals, whereas the β-casein is similar to the human protein, and displayed a section of unique AA composition and additional hydrophilic regions compared with bovine caseins. Elephant milk is destabilized by 62% alcohol, and it is speculated that the β-casein characteristics may allow maintenance of the colloidal nature of the casein micelle, a role that was previously only associated with κ-casein. The oligosaccharide content of milk was reported to be low in dairy animals but high in some other species such as humans and elephants. In the milk of the African elephant, lactose and oligosaccharides both occur at high levels. These levels are typically related to the content of α-LA in the mammary gland and thus point to a specialized carbohydrate synthesis, where the whey protein α-LA plays a role. We report the characterization of African elephant α-LA. Homology modeling of the α-LA showed that it is structurally similar to crystal structures of other mammalian species, which in turn may be an indication that its functional

  11. Rat beta casein cDNA: sequence analysis and evolutionary comparisons.

    OpenAIRE

    Blackburn, D E; Hobbs, A A; Rosen, J. M.

    1982-01-01

    The complete sequence of a 1072 nucleotide rat beta-casein cDNA insertion in the hybrid plasmid pC beta 23 has been determined. Primer extension was employed to determine the sequence of an additional 82 5'-terminal nucleotides in beta-casein mRNA. Rat beta-casein mRNA consists of a 696 nucleotide coding region, flanked by 52 nucleotide 5' and 406 nucleotide 3' noncoding regions, including a 40 nucleotide poly(A) tail. The derived 216 amino acid sequence of rat beta-casein was compared to the...

  12. Effect of caseins structure on the diffusion of solutes in caseins solutions and gels as studied by pulsed field gradient NMR

    OpenAIRE

    Le Feunteun, S.; Mariette, F.

    2006-01-01

    Caseins are milk's major proteins. They are present in milk as a suspension of particles called casein micelles. In the dairy industry, two classical processes which are acid and rennet coagulation or a combination of both are used to convert milk into gels, such as yoghurt or cheese curd. Acids gels are obtained through the acidification of the medium whereas rennet gels are formed after enzymatic hydrolysis. It is widely accepted that casein gels are so-called particle gels (Lucey J.A. J. D...

  13. Protein phosphatases active on acetyl-CoA carboxylase phosphorylated by casein kinase I, casein kinase II and the cAMP-dependent protein kinase

    International Nuclear Information System (INIS)

    The protein phosphatases in rat liver cytosol, active on rat liver acetyl-CoA carboxylase (ACC) phosphorylated by casein kinase I, casein kinase II and the cAMP-dependent protein kinase, have been partially purified by anion-exchange and gel filtration chromatography. The major phosphatase activities against all three substrates copurify through fractionation and appear to be identical to protein phosphatases 2A1 and 2A2. No unique protein phosphatase active on 32P-ACC phosphorylated by the casein kinases was identified

  14. Predictors of High Serum Casein Antibody Levels among Malnourished Infants and Young Children with Congenital Heart Disease

    Directory of Open Access Journals (Sweden)

    Inas R. El-Alameey

    2015-03-01

    CONCLUSION: Serum casein antibody levels play a significant role in the pathogenesis of malnutrition. Encouragement of breast feeding and avoidance of early cow's milk consumption could prevent the development of antibody response to bovine casein.

  15. A DIGITAL TOOL FOR CASEIN ISOLECTRIC POINT DETERMINATION

    Directory of Open Access Journals (Sweden)

    V. M.T. Trindade

    2015-08-01

    Full Text Available Introduction: Casein is a milk protein that has the same number of positive and negative charges in a pH around 4.7. This characteristic is called the isoelectric point (pI. The pI varies from protein to protein and it depends on the charges of the lateral chain of the constituent aminoacids. At this pH value the protein has its minimum solubility since the net charge is zero and the repulsion between molecules is decreased. Furthermore, the electrostatic interaction occurs between the protein molecules. Thus, they form clumps which tend to precipitate. On the other hand, when they are placed in a solution whose pH is above or below of its pI, the protein molecules have respectively a negative or positive net charge, with strong repulsion between themselves and great interaction with the solvent (water. Objective: This learning object presents a simulation of a laboratory practice for the determination of casein isoelectric point. Materials and Methods: Cartoons were planned in order to show the methodology procedures and biochemical fundamentals. Animations were developed with the aid of the Adobe ® Flash 8 software associated with logic programming. Results: The simulation consists of six steps that reproduce the activities performed in the laboratory. Among them, the user can observe different degrees of turbidity and / or precipitation of casein in solutions with different pH values, he can measure these values with paper indicator strips and then determine the pI of this protein. Conclusions: This learning object was tested by students of Biochemistry I, Pharmacy course since 2012/2. The navigation features, design, interaction, interactivity were considered excellent by 80% of students indicating that this object can be used as an interesting tool to assist the teaching and learning of basic biochemistry.  Available at:www.ufrgs.br/gcoeb/PontoIsoleletricoDaCaseina/PontoIsoleletricoDaCaseina.swf

  16. Enhanced casein kinase II activity in human tumour cell cultures

    DEFF Research Database (Denmark)

    Prowald, K; Fischer, H; Issinger, O G

    1984-01-01

    Casein kinase II (CKII) activity is enhanced as much as 2-3 fold in established and 4-5-fold in transformed human cell lines when compared to that of fibroblasts and primary human tumour cell cultures where CKII activity never exceeded a basic level. The high activity of CKII in transformed cells...... and in established cell lines was reduced to about the same basic level after treatment with heparin, a highly specific inhibitor of CKII activity. The activity of the cAMP-dependent protein kinase was virtually the same in fibroblasts and various human tumour cell lines investigated....

  17. Potential of casein kinase I in digestive cancer screening

    Directory of Open Access Journals (Sweden)

    Cristina Modak

    2009-10-01

    Full Text Available Casein kinase I is a group of ubiquitous Serine/Threonine kinases that have been implicated in both normal cellular functions and several pathological conditions including Alzheimer’ s disease and cancer. Recent findings in colon and pancreatic cancer have brought tremendous attention to these molecules as potential therapeutic targets in treatment of digestive cancers. In this review, we summarize up to date what is known about this family of kinases and their involvement in carcinogenesis and other pathological conditions. Our emphasis is on their implications in digestive cancers and their potential for cancer screening and therapy.

  18. Expression of casein kinase 2 during mouse embryogenesis

    DEFF Research Database (Denmark)

    Mestres, P; Boldyreff, B; Ebensperger, C; Hameister, H; Issinger, O G

    1994-01-01

    This paper deals with the expression and distribution of casein kinase 2 (CK-2) subunits in mouse embryos at different developmental stages. Expression was investigated at the mRNA level of CK-2 alpha- and beta-subunits by in situ hybridization and distribution at the protein level by...... immunohistochemistry using CK-2-alpha- and CK-2-beta-specific antibodies, respectively. In general both methods gave similar results. In earlier stages of mouse embryonic development (day 10.5 after coitus) CK-2 was more expressed in neuroepithelia than in all other tissues. From day 11.5 after coitus on, high...

  19. A DIGITAL TOOL FOR CASEIN ISOLECTRIC POINT DETERMINATION

    OpenAIRE

    V.M.T. Trindade; I. B. Santos; G. Zanatta; P. R. Arantes; C. G. Salbego

    2015-01-01

    Introduction: Casein is a milk protein that has the same number of positive and negative charges in a pH around 4.7. This characteristic is called the isoelectric point (pI). The pI varies from protein to protein and it depends on the charges of the lateral chain of the constituent aminoacids. At this pH value the protein has its minimum solubility since the net charge is zero and the repulsion between molecules is decreased. Furthermore, the electrostatic interaction occurs between the prote...

  20. Detecting β-Casein Variation in Bovine Milk

    OpenAIRE

    Anna Maria Caroli; Salvatore Savino; Omar Bulgari; Eugenio Monti

    2016-01-01

    In bovine species, β-casein (β-CN) is characterized by genetic polymorphism. The two most common protein variants are β-CN A2 (the original one) and A1, differing from A2 for one amino acid substitution (Pro67 to His67). Several bioactive peptides affecting milk nutritional properties can originate from β-CN. Among them, β-casomorphin-7 (BCM7) ranging from amino acid 60 to 66 can be released more easily from β-CN variants carrying His67 (A1 type) instead of Pro67 (A2 type). Nowadays, “A2 milk...

  1. The membrane-associated form of as1- casein interacts with cholesterol-rich detergent-resistant microdomains

    OpenAIRE

    Le Parc, Annabelle; Honvo Houeto, Edith; Pigat, Natascha; Chat, Sophie; Léonil, Joelle; Chanat, Eric

    2014-01-01

    Caseins, the main milk proteins, interact with colloidal calcium phosphate to form the casein micelle. The mesostructure of this supramolecular assembly markedly influences its nutritional and technological functionalities. However, its detailed molecular organization and the cellular mechanisms involved in its biogenesis have been only partially established. There is a growing body of evidence to support the concept that αs1-casein takes center stage in casein micelle building and transport ...

  2. Inhibition of hydroxyapatite growth by casein, a potential salivary phosphoprotein homologue.

    Science.gov (United States)

    Romero, Maria J R H; Nakashima, Syozi; Nikaido, Toru; Ichinose, Shizuko; Sadr, Alireza; Tagami, Junji

    2015-08-01

    Salivary phosphoproteins are essential in tooth mineral regulation but are often overlooked in vitro. This study aimed to evaluate the effect of casein, as a salivary phosphoprotein homologue, on the deposition and growth of hydroxyapatite (HA) on tooth surfaces. Hydroxyapatite growth was quantified using seeded crystal systems. Artificial saliva (AS) containing HA powder and 0, 10, 20, 50, or 100 μg ml(-1) of casein, or 100 μg ml(-1) of dephosphorylated casein (Dcasein), was incubated for 0-8 h at 37°C, pH 7.2. Calcium concentrations were measured using atomic absorption spectroscopy (AAS). Surface precipitation of HA on bovine enamel and dentine blocks, incubated in similar conditions for 7 d, was examined using field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) with selected area electron diffraction (SAED). Casein adsorption was assessed using modified Lowry assays and zeta-potential measurements. The AAS results revealed a concentration-dependent inhibition of calcium consumption. Hydroxyapatite precipitation occurred when no casein was present, whereas precipitation of HA was apparently completely inhibited in casein-containing groups. Adsorption data demonstrated increasingly negative zeta-potential with increased casein concentration and an affinity constant similar to proline-rich proteins with Langmuir modelling. Casein inhibited the deposition and growth of HA primarily through the binding of esterized phosphate to HA active sites, indicating its potential as a mineral-regulating salivary phosphoprotein homologue in vitro. PMID:26083784

  3. Rapid method for measuring protease activity in milk using radiolabeled casein

    International Nuclear Information System (INIS)

    A rapid means to detect the presence of protease activity in raw milk could be useful in predicting keeping ability of products made from that milk. A 30-min assay has been developed and compared with three other methods of detecting protease. Casein, [methyl-14C]-methylated-alpha was purchased from a radioisotope supplier. Concentrations of substrate from 2 to 20 nCi gave counts per minute, which increased linearly when counted with the Charm analyzer. There was not a significant difference in counting times of 10, 20, or 30 min. A mixture of sodium acetate and acetic acid precipitated nonhydrolyzed substrate with an efficiency of 97%. Comparison of the [14C] casein assay, a casein fluorescein isothiocyanate assay, trinitrobenzenesulfonic acid procedure, and the Hull procedure using protease from psychrotrophic bacteria revealed that the [14C] casein and casein fluorescein isothiocyanate methods were roughly equivalent and that the radiometric procedure was 10 times more sensitive than the trinitrobenzenesulfonic acid assay. The radiometric procedure was approximately 10(4) times more sensitive than the Hull procedure. The [14C] casein and casein fluorescein isothiocyanate methods were similar in time required, about 30 min, while the trinitrobenzenesulfonic acid assay and Hull method required about 1 h plus reagent preparation time. The [14C] casein procedure was most expensive per test; the other three were cheaper and similar to each other in cost

  4. Foams and surface rheological properties of β-casein, gliadin and glycinin

    NARCIS (Netherlands)

    Bos, M.A.; Dunnewind, B.; Vliet, T. van

    2003-01-01

    Interfacial rheological properties and their suitability for foam production and stability of two vegetable proteins were studied and compared to β-casein. Proteins used ranged from flexible to rigid/globular in the order of β-casein, gliadin and soy glycinin. Experiments were performed at pH 6.7. N

  5. Foams and surface rheological properties of b-casein, gliadin and glycinin

    NARCIS (Netherlands)

    Bos, M.A.; Dunnewind, B.; Vliet, van T.

    2003-01-01

    Interfacial rheological properties and their suitability for foam production and stability of two vegetable proteins were studied and compared to ß-casein. Proteins used ranged from flexible to rigid/globular in the order of ß-casein, gliadin and soy glycinin. Experiments were performed at pH 6.7. N

  6. Behaviour of casein micelles at conditions comparable to those in ice cream

    NARCIS (Netherlands)

    Jonkman, M.J.

    2000-01-01

    The physical properties of ice cream are mainly determined by the processing and the ingredients. Milk (powder) is one of the ingredients and ice cream thus contains casein, the major milk protein. A large proportion of casein in ice cream is present in the plasma phase of ice cream. Since the behav

  7. Purification and properties of a ribosomal casein kinase from rabbit reticulocytes

    DEFF Research Database (Denmark)

    Issinger, O G

    1977-01-01

    A casein kinase was isolated and purifed from rabbit reticulocytes. About 90% of the enzyme activity co-sedimented with the ribosomal fraction, whereas about 10% of the enzyme activity was found in the ribosome-free supernatant. Both casein kinases (the ribosome-bound enzyme as well as the free...

  8. The association of low-molecular-weight hydrophobic compounds with native casein micelles in bovine milk.

    Science.gov (United States)

    Cheema, M; Mohan, M S; Campagna, S R; Jurat-Fuentes, J L; Harte, F M

    2015-08-01

    The agreed biological function of the casein micelles in milk is to carry minerals (calcium, magnesium, and phosphorus) from mother to young along with amino acids for growth and development. Recently, native and modified casein micelles were used as encapsulating and delivery agents for various hydrophobic low-molecular-weight probes. The ability of modified casein micelles to bind certain probes may derive from the binding affinity of native casein micelles. Hence, a study with milk from single cows was conducted to further elucidate the association of hydrophobic molecules into native casein micelles and further understand their biological function. Hydrophobic and hydrophilic extraction followed by ultraperformance liquid chromatography-high resolution mass spectrometry analysis were performed over protein fractions obtained from size exclusion fractionation of raw skim milk. Hydrophobic compounds, including phosphatidylcholine, lyso-phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, showed strong association exclusively to casein micelles as compared with whey proteins, whereas hydrophilic compounds did not display any preference for their association among milk proteins. Further analysis using liquid chromatography-tandem mass spectrometry detected 42 compounds associated solely with the casein-micelles fraction. Mass fragments in tandem mass spectrometry identified 4 of these compounds as phosphatidylcholine with fatty acid composition of 16:0/18:1, 14:0/16:0, 16:0/16:0, and 18:1/18:0. These results support that transporting low-molecular-weight hydrophobic molecules is also a biological function of the casein micelles in milk. PMID:26074238

  9. MACROSTRUCTURE AND VISCOSITY OF AGGREGATING COLLOIDAL CASEIN MICELLES UNDER STRONG SHEARING FORCE

    Science.gov (United States)

    Casein particle gels are the main constituents in many classes of soft food colloids. In this study, we examined the stability of casein micelles under strong hydrodynamic forces, and investigated the effect of continuous applied shear on the coagulation properties of rennet-induced milk. At the a...

  10. Effects of gamma-irradiation on some properties of bovine casein micelles

    International Nuclear Information System (INIS)

    Sedimentation studies and electron microscopic observations revealed that an association between casein micelles dispersed in water or milk serum was not induced significantly by gamma-irradiation of exposure up to 3 x 106R, whereas a release of nonprotein nitrogen was observed to a certain extent. It was concluded from the results of turbidi-metry and gel filtration using 3 size groups of casein micelles, namely large, medium and small, that an irradiation-induced polymerization or association occurred within individual casein micelles, and strengthend the micelle structure. Thus the irradiated casein micelles resisted, more or less, to the solubilizing effect of NaCl, EDTA, pyrophosphate and urea. Stabilities of casein micelles for ethanol and for acidification to an isoelectric point were decreased and increased, respectively, after irradiation. Gamma irradiation also caused the decrease of glycomacropeptide released from casein micelles by the action of rennin, and this resulted in the delay of rennin-coagulation of casein. There were no essential differences among the 3 size groups of casein micelles concerning the above described tendencies. (auth.)

  11. Effect of Fluoride, Casein Phosphopeptide–Amorphous Calcium Phosphate and Casein Phosphopeptide–Amorphous Calcium Phosphate Fluoride on Enamel Surface Microhardness After Microabrasion: An In Vitro Study

    OpenAIRE

    Ghazaleh Ahmadi Zenouz; Fariba Ezoji; Seyede Anese Enderami; Soraya Khafri

    2016-01-01

    Objectives: This study aimed to assess the effect of applying casein phosphopeptide–amorphous calcium phosphate (CPP-ACP) paste, casein phosphopeptide–amorphous calcium phosphate fluoride (CPP-ACPF) paste and sodium fluoride gel on surface microhardness of enamel after microabrasion.Materials and Methods: Thirty freshly extracted human premolars were selected. All samples were subjected to hardness indentations made with the Vickers hardness machine and the average value was recorded as the i...

  12. beta-Casein mRNA sequesters a single-stranded nucleic acid-binding protein which negatively regulates the beta-casein gene promoter.

    OpenAIRE

    Altiok, S; Groner, B

    1994-01-01

    beta-Casein gene expression in mammary epithelial cells is under the control of the lactogenic hormones, glucocorticoids, insulin, and prolactin. The hormonal control affects gene transcription, and several regulatory elements in the beta-casein gene promoter between positions -80 and -221 have previously been identified. A region located in the promoter between positions -170 and -221 contains overlapping sequences for negative and positive regulatory elements. A sequence-specific single-str...

  13. PFG-NMR self-diffusion in casein dispersions: effect of probe size and protein aggregate size

    NARCIS (Netherlands)

    Salami, S.; Rondeau, C.; Duynhoven, van J.P.M.; Mariette, F.

    2013-01-01

    The self-diffusion coefficients of different molecular weight PEGs (Polyethylene glycol) and casein particles were measured, using a pulsed-gradient nuclear magnetic resonance technique (PFG-NMR), in native phosphocaseinate (NPC) and sodium caseinate (SC) dispersions where caseins are not structured

  14. Chemical synthesis and structure elucidation of bovine κ-casein (1-44)

    International Nuclear Information System (INIS)

    The caseins (αs1, αs2, β, and κ) are phosphoproteins present in bovine milk that have been studied for over a century and whose structures remain obscure. Here we describe the chemical synthesis and structure elucidation of the N-terminal segment (1-44) of bovine κ-casein, the protein which maintains the micellar structure of the caseins. κ-Casein (1-44) was synthesised by highly optimised Boc solid-phase peptide chemistry and characterised by mass spectrometry. Structure elucidation was carried out by circular dichroism and nuclear magnetic resonance spectroscopy. CD analysis demonstrated that the segment was ill defined in aqueous medium but in 30% trifluoroethanol it exhibited considerable helical structure. Further, NMR analysis showed the presence of a helical segment containing 26 residues which extends from Pro8 to Arg34. This is First report which demonstrates extensive secondary structure within the casein class of proteins

  15. Hydrolyzed casein reduces diet-induced obesity in male C57BL/6J mice

    DEFF Research Database (Denmark)

    Lillefosse, Haldis Haukås; Tastesen, Hanne Sørup; Du, Zhen-Yu;

    2013-01-01

    The digestion rate of dietary protein is a regulating factor for postprandial metabolism both in humans and animal models. However, few data exist about the habitual consumption of proteins with different digestion rates with regard to the development of body mass and diet-induced obesity. Here, we...... used a factorial ANOVA design to investigate the effects of protein form (intact vs. hydrolyzed casein) and protein level (16 vs. 32 energy percent protein) on body mass gain and adiposity in obesity-prone male C57BL/6J mice fed Western diets with 35 energy percent fat. Mice fed the hydrolyzed casein...... diets had higher spontaneous locomotor activity than mice fed intact casein. During the light phase, mice fed hydrolyzed casein tended (P = 0.08) to have a lower respiratory exchange ratio, indicating lower utilization of carbohydrates as energy substrate relative to those fed intact casein. In further...

  16. Dissolution and enzymatic hydrolysis of casein micelles studied by dynamic light scattering

    Institute of Scientific and Technical Information of China (English)

    LIU Rui; QI Wei; SU Rongxin; ZHANG Yubin; JIN Fengmin; HE Zhimin

    2007-01-01

    The effects of temperature,ionic strength,and enzymatic hydrolysis on the average hydrodynamic radius (Rh) of casein micelles in phosphate buffer were studied by using dynamic light scattering.The results showed that the average Rh value of casein mieelles decreased irreversibly during the heating,decreased with the increase of ionic strength in lower ionic strength solution (less than 0.05 tool/L),but opposite in higher ionic strength solution (above 0.1 tool/L).The Rh value of casein increased rapidly during the process of enzymatic hydrolysis,and the structural model of casein micelles in the enzymatic hydrolysis process was also proposed,i.e.the casein micelle changed from compact sphere into unfolded and regularly flocky peptides.

  17. Rheology of Sodium Caseinate Stabilized Oil-in-Water Emulsions

    Science.gov (United States)

    Dickinson; Golding

    1997-07-01

    We report on shear rheological measurements at 30°C of fine oil-in-water emulsions (volume-surface average diameter tetradecane as the dispersed phase (10, 35, or 45 vol%). Strong sensitivity of rheological behavior to total protein concentration was indicated by both steady-state viscometry and small-deformation oscillatory experiments. The behavior can be classified into three types, depending on the protein/oil ratio. (1) Emulsions containing insufficient protein for (near-) saturation protein surface coverage develop a time-dependent increase in low-stress apparent viscosity and associated shear-thinning behavior; this can be attributed to bridging flocculation. (2) Emulsions having full protein surface coverage but relatively little excess unadsorbed protein in the continuous phase are stable Newtonian liquids. (3) Emulsions containing a substantial excess of unadsorbed sodium caseinate exhibit considerable pseudoplasticity which can be attributed to depletion flocculation. Taken as a whole, the time-dependent rheological properties for this set of emulsions as a function of protein content and oil volume fraction are largely consistent with our previous results on the creaming stability and the particle gel microstructure for these same emulsion systems. In particular, the reversible flocculation of emulsion samples of high protein content is readily explicable in terms of depletion flocculation of droplets by unadsorbed protein existing in the form of approximately spherical caseinate submicelles. PMID:9241217

  18. Casein Phosphopeptide-Amorphous Calcium Phosphate Nanocomplexes: A Structural Model.

    Science.gov (United States)

    Cross, Keith J; Huq, N Laila; Reynolds, Eric C

    2016-08-01

    Tryptic digestion of the calcium-sensitive caseins yields casein phosphopeptides (CPP) that contain clusters of phosphorylated seryl residues. The CPP stabilize calcium and phosphate ions through the formation of complexes. The calcium phosphate in these complexes is biologically available for intestinal absorption and remineralization of subsurface lesions in tooth enamel. We have studied the structure of the complexes formed by the CPP with calcium phosphate using a variety of nuclear magnetic resonance (NMR) techniques. Translational diffusion measurements indicated that the β-CN(1-25)-ACP nanocomplex has a hydrodynamic radius of 1.526 ± 0.044 nm at pH 6.0, which increases to 1.923 ± 0.082 nm at pH 9.0. (1)H NMR spectra were well resolved, and (3)JH(N)-H(α) measurements ranged from a low of 5.5 Hz to a high of 8.1 Hz. Total correlation spectroscopy and nuclear Overhauser effect spectroscopy spectra were acquired and sequentially assigned. Experiments described in this paper have allowed the development of a structural model of the β-CN(1-25)-amorphous calcium phosphate nanocomplex. PMID:27434168

  19. Antioxidant activity and bioaccessibility of phenols-enriched edible casein/caseinate coatings during in vitro digestion.

    Science.gov (United States)

    Helal, Ahmed; Desobry, Stephane; Banon, Sylvie; Shamsia, Sherif M

    2015-02-01

    Active films were developed for food coating applications. Entrapped phenol susceptibility to digestion was studied. Sodium caseinate (Na-CN) coatings were formulated with 0, 10, 20% Casein (CN) incorporating selected phenols as model antioxidants. This study investigated phenol/CN/Na-CN interactions, in vitro bioaccessibility of phenols and CN role in phenols retention during in vitro gastric and pancreatic digestion. The antioxidant activity of catechin (CAT), rutin (RUT), chlorogenic acid (CHL), gallic acid (GAL), and tannic acid (TA) in coatings varied with the phenolic compound type and CN concentration and was related to phenol hydrophobic binding to CN. ABTS method gave activities ranged from 412 down to 213, and DPPH method gave values from 291·7 to 190·9. An inverse relationship was found with CN content due to CN/phenol interaction. During digestion, a part of phenols was degraded by alkaline pH of pancreatic fluid. Simultaneously, CN proteolysis led to release of phenols and the bioaccessibility index remained above 80% for all phenols. The results suggested the possibility of protecting phenols against oxidation and digestive alteration by entrapment in CN and Na-CN coating films. These positive results showed the ability to produce antioxidant-enriched edible coatings to increase food protection and phenol nutritional intake. PMID:25327452

  20. Solution and film properties of sodium caseinate/glycerol and sodium caseinate/polyethylene glycol edible coating systems.

    Science.gov (United States)

    Siew, D C; Heilmann, C; Easteal, A J; Cooney, R P

    1999-08-01

    The aim of this study is to determine the effects of plasticizer hydrogen bonding capability and chain length on the molecular structure of sodium caseinate (NaCAS), in NaCAS/glycerol and NaCAS/polyethylene glycol 400 (PEG) systems. Both solution and film phases were investigated. Glycerol and PEG reduced the viscosity of aqueous NaCAS, with the latter having a greater effect. This was explained in terms of protein/plasticizer aggregate size and changes to the conformation of the caseinate chain. In the film phase, glycerol caused more pronounced changes to the film tensile strength compared with PEG. However, the effect of glycerol on film water vapor permeability was smaller. These observations are attributed to the differences in plasticizer size and hydrogen bonding strength that controls the protein-plasticizer and protein-protein interactions in the films. Glass transition calculations from the tensile strength data indicate that the distribution of bonding interactions is more homogeneous in NaCAS/PEG films than in NaCAS/glycerol films. PMID:10552668

  1. Spectrofluoremetric and molecular docking study on the interaction of bisdemethoxycurcumin with bovine β-casein nanoparticles

    International Nuclear Information System (INIS)

    The interaction of bisdemethoxycurcumin (BDMC), as one of the main active component of turmeric (Curcuma longa L.), with bovine β-casein nanoparticle, as an efficient drug carrier system, was investigated using steady-state fluorescence spectroscopy and molecular docking calculations. Results of fluorescence quenching experiments, Forster energy transfer measurements and molecular docking calculations suggested that BDMC bind to the hydrophobic core of β-casein via formation of 3 hydrogen bonds and several vander Waals contacts that represented the encapsulation of BDMC in β-casein micelle nanoparticles. The binding parameters including number of substantive binding sites and the binding constants were evaluated by fluorescence quenching method. Additionally, the cytotoxicity of free BDMC and BDMC-β-casein complex in human breast cancer cell line MCF7 was evaluated in vitro. The study revealed the higher cytotoxic effects of encapsulated BDMC on MCF7 cells compared to equal dose of free BDMC. -- Highlights: • BDMC binds to the hydrophobic core of β-casein. • The effective encapsulation of BDMC in β-casein micelle nanoparticles was shown. • Enhanced cytotoxicity was observed for encapsulated BDMC in β-casein nanoparticles

  2. Nutritional evaluation of caseins and whey proteins and their hydrolysates from Protamex

    Institute of Scientific and Technical Information of China (English)

    SINDAYIKENGERA Séverin; XIA Wen-shui

    2006-01-01

    Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard.The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.

  3. Spectrofluoremetric and molecular docking study on the interaction of bisdemethoxycurcumin with bovine β-casein nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Mehranfar, Fahimeh [Department of Chemistry, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Bordbar, Abdol-Khalegh, E-mail: bordbar@chem.ui.ac.ir [Department of Chemistry, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Keyhanfar, Mehrnaz; Behbahani, Mandana [Faculty of Advanced Sciences and Technologies, Department of Biotechnology, University of Isfahan, Isfahan, 81746-73441 (Iran, Islamic Republic of)

    2013-11-15

    The interaction of bisdemethoxycurcumin (BDMC), as one of the main active component of turmeric (Curcuma longa L.), with bovine β-casein nanoparticle, as an efficient drug carrier system, was investigated using steady-state fluorescence spectroscopy and molecular docking calculations. Results of fluorescence quenching experiments, Forster energy transfer measurements and molecular docking calculations suggested that BDMC bind to the hydrophobic core of β-casein via formation of 3 hydrogen bonds and several vander Waals contacts that represented the encapsulation of BDMC in β-casein micelle nanoparticles. The binding parameters including number of substantive binding sites and the binding constants were evaluated by fluorescence quenching method. Additionally, the cytotoxicity of free BDMC and BDMC-β-casein complex in human breast cancer cell line MCF7 was evaluated in vitro. The study revealed the higher cytotoxic effects of encapsulated BDMC on MCF7 cells compared to equal dose of free BDMC. -- Highlights: • BDMC binds to the hydrophobic core of β-casein. • The effective encapsulation of BDMC in β-casein micelle nanoparticles was shown. • Enhanced cytotoxicity was observed for encapsulated BDMC in β-casein nanoparticles.

  4. Antiproliferative activity of tea catechins associated with casein micelles, using HT29 colon cancer cells.

    Science.gov (United States)

    Haratifar, S; Meckling, K A; Corredig, M

    2014-02-01

    Numerous studies have shown that green tea polyphenols display anticancer activities in many organ sites by using different experimental models in rodents and in cultured cell lines in vitro. The present study tested the ability of casein micelles to deliver biologically active concentrations of polyphenols to HT-29 colon cancer cells. Epigallocatechin gallate (EGCG), the major catechin found in green tea, was used as the model molecule, as it has been shown to have antiproliferative activity on colon cancer cells. In the present work, we hypothesized that due to the binding of caseins with EGCG, casein micelles may be an ideal platform for the delivery of this bioactive molecule and that the binding would not affect the bioaccessibility of EGCG. The cytotoxicity and proliferation behavior of HT-29 colon cancer cells when exposed to free EGCG was compared with that of nanoencapsulated EGCG in casein micelles of skim milk. Epigallocatechin gallate-casein complexes were able to decrease the proliferation of HT-29 cancer cells, demonstrating that bioavailability may not be reduced by the nanoencapsulation. As casein micelles may act as protective carriers for EGCG in foods, it was concluded that nanoencapsulation of tea catechins in casein micelles may not diminish their antiproliferative activity on colon cancer cells compared with free tea catechins. PMID:24359816

  5. A novel method for separation of caseins from milk by phosphates precipitation.

    Science.gov (United States)

    Yen, Chon-Ho; Lin, Yin-Shen; Tu, Ching-Fu

    2015-01-01

    Milk protein of farm animals is difficult to isolate because of the presence of casein micelles, which are hard to separate from whey by using centrifugation or filtration. Insoluble casein micelles also create an obstacle for purification instruments to operate efficiently. The conventional method, to precipitate caseins by lowering pH to 4.6 and then recover the whey fraction for further purification using chromatography techniques, is not applicable to proteins having an isoelectric point similar to caseins. In addition, the acid condition used for casein removal usually leads to significantly poor yields and reduced biological activities. In this study, a novel method of precipitating caseins under neutral or weak acidic conditions is presented. The method employs a phosphate salt and a freeze-thaw procedure to obtain a casein-free whey protein fraction. This fraction contains more than 90% yield with little loss of bioactivity of the target protein, and is readily available for further chromatographic purification. This method was successfully applied to purify recombinant human factor IX and recombinant hirudin from the milk of transgenic pigs in the presented study. It is an efficient pretreatment approach prior to chromatographic purification of milk protein from farm animals and particularly of great value to collect those recombinants secreted from transgenic livestock. PMID:24372141

  6. A new tetra-primer ARMS-PCR for genotyping bovine kappa-casein polymorphisms.

    Science.gov (United States)

    Fonseca, P A S; Rosse, I C; Demiranda, M; Machado, M A; Verneque, R S; Peixoto, M G C D; Carvalho, M R S

    2013-01-01

    Kappa-casein (κ-casein) is one of the most abundant milk proteins. Its main function is to avoid the aggregation of casein micelles, keeping them, and therefore calcium phosphate, in pockets in solution. In bovines, a κ-casein functional polymorphism has been associated with fat, calcium, and protein milk contents and faster curd contraction in cheese production. Quicker curd contraction reduces the loss of milk solids, enhancing cheese yield. This polymorphism induces a double amino acid substitution (Thr136Ile and Ala148Asp). The polymorphism is normally detected by PCR-RFLP, which is a laborious method. An interesting methodological alternative is the tetra-primer amplification refractory mutation system PCR (tetra-primer ARMS-PCR). A tetra-primer ARMS-PCR for the detection of this κ-casein polymorphism has been described. However, specificity was not achieved, probably due to problems with primer design. We developed a new tetra-primer ARMS-PCR for the detection of the κ-casein polymorphism. This new method was validated in a double-blind test, by comparison with the results obtained for 50 Guzerá bulls formerly genotyped by PCR-RFLP. This new method achieved 100% sensitivity and specificity. We conclude that this method is a useful, cost-efficient alternative for the detection of functional κ-casein polymorphisms. PMID:24390998

  7. Reassociation of dissociated caseins upon acidification of heated pH-adjusted skim milk.

    Science.gov (United States)

    Anema, Skelte G; Li, Yuming

    2015-05-01

    Milk was heated at different pH (pH 6.5-7.1) and temperatures (20-120 °C/10 min). This resulted in different levels of casein and denatured whey proteins to be distributed between the colloidal and serum phases. The milks were subsequently acidified and the distribution of protein between colloidal and serum was monitored at different pH. On acidification to pH 5.4, the serum phase caseins and denatured whey proteins partially reassociated with the caseins, although a complex behaviour was observed at ∼ pH 5.4 where additional casein dissociation occurred in some samples. At pH below 5.4 the caseins and denatured whey proteins rapidly aggregated. No separate aggregation of κ-casein/denatured whey protein complexes or κ-casein depleted micelles was observed. The earlier gelation of milks heated at higher pH was likely to be due to the destabilisation of the entire milk protein system rather than a preferential aggregation of the serum phase proteins. PMID:25529690

  8. Temperature-dependent dynamics of dry and hydrated β-casein studied by quasielastic neutron scattering.

    Science.gov (United States)

    Dhindsa, Gurpreet K; Tyagi, Madhusudan; Chu, Xiang-qiang

    2014-09-18

    β-Casein is a component of casein micelle with amphillic nature and is recognized as a "natively disordered" protein that lacks secondary structures. In this study, the temperature and hydration effects on the dynamics of β-casein are explored by quasielastic neutron scattering (QENS). An upturn in the mean square displacement (MSD) of hydrated β-casein indicates an increase of protein flexibility at a temperature of ~225 K. Another increase in MSD at ~100 K, observed in both dry and hydrated β-casein, is ascribed to the methyl group rotations, which are not sensitive to hydration. QENS analysis in the energy domain reveals that the fraction of hydrogen atoms participating in motion in a sphere of diffusion is highly hydration dependent and increases with temperature. In the time domain analysis, a logarithmic-like decay is observed in the range of picosecond to nanosecond (β-relaxation time) in the dynamics of hydrated β-casein. This dynamical behavior has been observed in hydrated globular and oligomeric proteins. Our temperature-dependent QENS experiments provide evidence that lack of a secondary structure in β-casein results in higher flexibility in its dynamics and easier reversible thermal unfolding compared to other rigid biomolecules. PMID:25144497

  9. The Effect of Milk Constituents and Crowding Agents on Amyloid Fibril Formation by κ-Casein.

    Science.gov (United States)

    Liu, Jihua; Dehle, Francis C; Liu, Yanqin; Bahraminejad, Elmira; Ecroyd, Heath; Thorn, David C; Carver, John A

    2016-02-17

    When not incorporated into the casein micelle, κ-casein, a major milk protein, rapidly forms amyloid fibrils at physiological pH and temperature. In this study, the effects of milk components (calcium, lactose, lipids, and heparan sulfate) and crowding agents on reduced and carboxymethylated (RCM) κ-casein fibril formation was investigated using far-UV circular dichroism spectroscopy, thioflavin T binding assays, and transmission electron microscopy. Longer-chain phosphatidylcholine lipids, which form the lining of milk ducts and milk fat globules, enhanced RCM κ-casein fibril formation irrespective of whether the lipids were in a monomeric or micellar state, whereas shorter-chain phospholipids and triglycerides had little effect. Heparan sulfate, a component of the milk fat globule membrane and catalyst of amyloid deposition in extracellular tissue, had little effect on the kinetics of RCM κ-casein fibril formation. Major nutritional components such as calcium and lactose also had no significant effect. Macromolecular crowding enhances protein-protein interactions, but in contrast to other fibril-forming species, the extent of RCM κ-casein fibril formation was reduced by the presence of a variety of crowding agents. These data are consistent with a mechanism of κ-casein fibril formation in which the rate-determining step is dissociation from the oligomer to give the highly amyloidogenic monomer. We conclude that the interaction of κ-casein with membrane-associated phospholipids along its secretory pathway may contribute to the development of amyloid deposits in mammary tissue. However, the formation of spherical oligomers such as casein micelles is favored over amyloid fibrils in the crowded environment of milk, within which the occurrence of amyloid fibrils is low. PMID:26807595

  10. Localization of the casein gene family to a single mouse chromosome

    OpenAIRE

    1982-01-01

    A series of mouse-hamster somatic cell hybrids containing a variable number of mouse chromosomes and a constant set of hamster chromosomes have been used to determine the chromosomal location of a family of hormone-inducible genes, the murine caseins. Recombinant mouse cDNA clones encoding the alpha-, beta-, and gamma-caseins were constructed and used in DNA restriction mapping experiments. All three casein cDNAs hybridized to the same set of somatic cell hybrid DNAs isolated from cells conta...

  11. Molecular mechanisms involved in casein gene expression and secretion in mouse mammary epithelial cells

    International Nuclear Information System (INIS)

    Mouse mammary epithelial cells (MMEC) secrete a group of milk-specific proteins including various caseins and whey proteins. Dissociated mammary epithelial cells maintain expression of most of their differentiated functions only if cells are plated on a suitable substratum. Casein production and section, cell morphology, and production of α-lactalbumin have been used as markers to assess the degree of differentiation of mammary cells in culture. The general consensus is that cells express their differentiated properties at high levels and for longer periods of time on such substrata. In this paper, the authors demonstrate that modulation of the expression of caseins by floating collagen gels is manifested at several regulatory points

  12. Search for phosphopeptides in the feces of axenic rats fed radioactive ovine casein

    International Nuclear Information System (INIS)

    Radioactive ovine casein was obtained by injecting 100 μCi of 14C-Ser into the jugular vein of an ewe. The milk collected 17 and 24 h after this injection contained 12% of the radioactivity injected in protein form. The seryl residues were specificially labelled. This casein was used as the only protein source fed to axenic rats; 0.30% of the tracer ingested was found in the feces of those rats. Since phosphoserine represented 25% of the total casein seryl residues, the phosphopeptides may not be selectively unabsorbable

  13. Proteolysis of ovine and caprine Caseins in solution by enzymatic extracts from flowers of Cynara cardunculuso

    OpenAIRE

    Sousa, M. José; Malcata, F. Xavier

    1998-01-01

    Primary proteolysis of ovine and caprine Na-caseinate at 30°C in phosphate buffer at pH 6.5 or 5.5 in the absence of NaCl and at pH 5.2 with 5% (w/v) NaCl by cardosins in aqueous extracts of Cynara cardunculus flowers was investigated using urea-polyacrylamide gel electrophoresis and reversed-phase high performance liquid chromatography. Caprine caseinate underwent more extensive degradation than ovine caseinate under the same conditions (pH 6.5 and pH 5.5); proteolysis of b- and ...

  14. Pressure-induced dissociation of casein micelles: size distribution and effect of temperature

    Directory of Open Access Journals (Sweden)

    Gebhardt R.

    2005-01-01

    Full Text Available Pressure-induced dissociation of a turbid solution of casein micelles was studied in situ in static and dynamic light scattering experiments. We show that at high pressure casein micelles decompose into small fragments comparable in size to casein monomers. At intermediate pressure we observe particles measuring 15 to 20 nm in diameter. The stability against pressure dissociation increased with temperature, suggesting enhanced hydrophobic contacts. The pressure transition curves are biphasic, compatible with a temperature (but not pressure-dependent conformational equilibrium of two micelle species. Our thermodynamic model predicts an increase in structural entropy with temperature.

  15. NMR study of the polyethyleneglycols diffusion in casein suspensions and gels

    OpenAIRE

    Colsenet, R.; Mariette, F.; Söderman, O.

    2006-01-01

    PFG-NMR spectroscopy was used to study the diffusion of molecular probes (polyethyleneglycols) in casein suspensions and gels in terms of the effects of probe molecular size (molecular mass between 1080 and 634000 g/mol), casein concentrations (from 3.24 to 16.22 g/100 g) and effects of rennet coagulation. A strong dependency of diffusion on probe size was observed, both in casein suspensions and in gels: as the PEG size increased, the diffusion was reduced. This effect was more pronoun...

  16. Calcium, vitamin D casein and whey protein intakes and periodontitis among Danish Adults

    DEFF Research Database (Denmark)

    Adegboye, Amanda Rodrigues Amorim; Boucher, Barbara J; Christensen, Lisa Bøge;

    2016-01-01

    OBJECTIVE: To investigate whether intakes of Ca, vitamin D, casein and whey are associated with periodontitis and to investigate the possibility of interactions between them. DESIGN: Cross-sectional study. An Internet-based, 267-item FFQ was used to assess dietary intake. Intakes of casein (32.0 g...... inversely associated with periodontitis. Consumption of foods rich in Ca, casein and whey (e.g. dairy foods) should be promoted, as they may contribute to the prevention of periodontitis. Further longitudinal studies are required to confirm these associations....

  17. β-casein as a functional food component

    DEFF Research Database (Denmark)

    Sulewska, Anna Maria

    Milk is a complex mixture of various components which include proteins, carbohydrates, lipids and minerals. Milk proteins can be divided into two classes: whey proteins (20%) and caseins (80% of total milk protein). As thermal treatment is a traditional method of food preservation, interactions...... which can improve stability and safety of food. Traditional methods of food preservation and sterilization require high temperature, which may result in undesirable changes such as change of color or texture. Development of novel food processing methods, which allow to create new milk-based products...... with novel textures, is highly valuable to the dairy industry. Understanding interactions that undergo during food processing is necessary for proper design of process. For example control of aggregation is required to achieve desired texture of the final product....

  18. Detecting β-Casein Variation in Bovine Milk

    Directory of Open Access Journals (Sweden)

    Anna Maria Caroli

    2016-01-01

    Full Text Available In bovine species, β-casein (β-CN is characterized by genetic polymorphism. The two most common protein variants are β-CN A2 (the original one and A1, differing from A2 for one amino acid substitution (Pro67 to His67. Several bioactive peptides affecting milk nutritional properties can originate from β-CN. Among them, β-casomorphin-7 (BCM7 ranging from amino acid 60 to 66 can be released more easily from β-CN variants carrying His67 (A1 type instead of Pro67 (A2 type. Nowadays, “A2 milk” is produced in different countries claiming its potential benefits in human health. The aim of this study was to further develop and apply an isoelectric focusing electrophoresis (IEF method to bulk and individual milk samples in order to improve its use for β-CN studies. We succeeded in identifying A2 milk samples correctly and quantifying the percentage of A2, A1, and B variants in bulk samples not derived from A2 milk as well as in individual milk samples. The method allows us to quantify the relative proportion of β-CN variants in whole milk without eliminating whey protein by acid or enzymatic precipitation of caseins. The aim of this study was also to study the different behavior of β-CN and β-lactoglobulin (β-LG in the presence of trichloroacetic acid (TCA. The higher sensitivity of β-CN to TCA allows quantifying β-CN variants after TCA fixation because β-LG is not visible. Monitoring β-CN variation in cattle breeds is important in order to maintain a certain balance between Pro67 and His67 in dairy products. Overall, the debate between A1 and A2 milk needs further investigation.

  19. Antioxidant activity of camel milk casein before and after in vitro simulated enzymatic digestion

    Directory of Open Access Journals (Sweden)

    Zeineb Jrad

    2014-11-01

    Full Text Available The effect of a successive in vitro hydrolysis by pepsin and pancreatin on the free radical scavenging activity of camel milk casein was investigated in order to assess the effect of gastro-intestinal digestion. Hydrolysis of camel casein was controlled by reversed-phase high performance liquid chromatography. Anti-oxidant activity was measured by the 2,2’-azino-bis-(3-ethylbensothiazoline-6- sulfonic acid (ABTS method. The Trolox equivalent antioxidant capacity (TEAC values of camel casein and its hydrolysate were 1.6±0.12 μmol TE/mg protein and 0.25 μmol TE/μmol eq. NH2, respectively. After digestion, the scavenging activity of the casein peptides was more efficient than those reported in the literature regarding digestive hydrolysates of camel milk, colostrum and whey proteins.

  20. Ultrasound effects on the assembly of casein micelles in reconstituted skim milk.

    Science.gov (United States)

    Liu, Zheng; Juliano, Pablo; Williams, Roderick P W; Niere, Julie; Augustin, Mary Ann

    2014-05-01

    Reconstituted skim milks (10 % w/w total solids, pH 6·7-8·0) were ultrasonicated (20, 400 or 1600 kHz at a specific energy input of 286 kJ/kg) at a bulk milk temperature of casein micelle in milk, with greater effects at higher pH and lower frequency. Low frequency ultrasound caused greater disruption of casein micelles causing release of protein from the micellar to the serum phase than high frequency. The released protein re-associated to form aggregates of smaller size but with surface charge similar to the casein micelles in the original milk. Ultrasound may be used as a physical intervention to alter the size of the micelles and the partitioning of caseins between the micellar and serum phases in milk. The altered protein equilibria induced by ultrasound treatment may have potential for the development of milk with novel functionality. PMID:24351847

  1. Enzymatically controlled material design with casein--from defined films to localized deposition of particles.

    Science.gov (United States)

    Strube, Oliver I; Rüdiger, Arne A; Bremser, Wolfgang

    2015-05-10

    A new concept for deposition and material design of coatings from biological compounds is presented. An enzymatic reaction triggers the specific coagulation of particles on a support surface. The first examined model system is casein and is based on the natural rennet reaction as applied in the process of cheese-making. The aspartic protease chymosin is immobilized on a support surface and cleaves the hydrophilic parts of the casein micelles, inducing deposition. The concept allows for a high level of control over film characteristics and enables the formation of site-specific film structures. The variability rages from formation of casein films with several micrometers film thickness to the targeted deposition of casein micelles. PMID:25456052

  2. Stimulation of muscle protein synthesis by whey and caseinate ingestion after resistance exercise in elderly individuals

    DEFF Research Database (Denmark)

    Dideriksen, K J; Reitelseder, S; Petersen, S G;

    2011-01-01

    Sarcopenia is a well-known phenomenon in elderly individuals and resistance exercise together with sufficient amino acid (AA) availability has proved to be a counteractive implement. However, the source of AA and supplement timing require further investigation. The objective was to compare muscle...... protein synthesis (MPS) to intakes of whey and caseinate after heavy resistance exercise in healthy elderly individuals, and, furthermore, to compare the timing effect of caseinate intake. Twenty-four elderly men and women (mean ± SEM; 68 ± 1 years) were randomized to one of four groups: caseinate intake...... before exercise (CasPre), caseinate intake immediately after exercise (CasPost), whey intake immediately after exercise (Whey), or intake of a non-caloric control drink (Control). Muscle myofibrillar and collagen fractional synthesis rates (FSR) were measured by a primed continuous infusion of L-[1...

  3. Irradiation effect on {alpha}- and {beta}-caseins of milk and Queso Blanco cheese determined by capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Ham, J.S.; Jeong, S.G.; Lee, S.G.; Han, G.S.; Chae, H.S.; Yoo, Y.M.; Kim, D.H. [Animal Food Processing Division, National Institute of Animal Science, Suwon 441-706 (Korea, Republic of); Lee, W.K. [College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Jo, C. [Department of Animal Science and Biotechnology, Chungnam National University, Daejeon 305-764 (Korea, Republic of)], E-mail: cheorun@cnu.ac.kr

    2009-02-15

    Milk and Queso Blanco cheese were exposed to irradiation with doses of 1, 2, 3, 5, and 10 kGy to investigate the irradiation effect on {alpha}- and {beta}-casein using a capillary electrophoresis. {alpha}{sub S1}-Casein to total protein ratio in raw milk was decreased from 19.63% to 8.64% by 10 kGy of gamma irradiation. The ratio of {alpha}{sub S1}- to {alpha}{sub S0}-casein was also decreased from 1.38 to 0.53, which showed {alpha}{sub S1}-casein is more susceptible to gamma irradiation than {alpha}{sub S0}-casein. Similarly, {alpha}{sub S1}-casein to total protein ratio in Queso Blanco cheese was decreased from 17.48% to 7.82% and the ratio of {alpha}{sub S1}- to {alpha}{sub S0}-casein was decreased from 1.16 to 0.43 by 10 kGy of gamma irradiation. Dose-dependent reduction of {beta}{sub A1}-casein was also found. {beta}{sub A1}-Casein to total protein ratios in raw milk and Queso Blanco cheese were decreased from 22.00% to 14.16% and from 21.96% to 13.89% after 10 kGy, respectively. The ratios of {beta}{sub A1}- to {beta}{sub A2}-casein were from 1.10 to 0.64 and 0.93 to 0.57 in milk and Queso Blanco cheese, respectively. However, {alpha}{sub S0}-, {beta}{sub B}-, and {beta}{sub A3}-casein increased by irradiation at 10 kGy. The results suggest that {alpha}{sub S1}-casein and {beta}{sub A1}-casein were more susceptible to gamma irradiation, and may be related to the reduction of milk allergenicity caused by gamma irradiation.

  4. Irradiation effect on α- and β-caseins of milk and Queso Blanco cheese determined by capillary electrophoresis

    International Nuclear Information System (INIS)

    Milk and Queso Blanco cheese were exposed to irradiation with doses of 1, 2, 3, 5, and 10 kGy to investigate the irradiation effect on α- and β-casein using a capillary electrophoresis. αS1-Casein to total protein ratio in raw milk was decreased from 19.63% to 8.64% by 10 kGy of gamma irradiation. The ratio of αS1- to αS0-casein was also decreased from 1.38 to 0.53, which showed αS1-casein is more susceptible to gamma irradiation than αS0-casein. Similarly, αS1-casein to total protein ratio in Queso Blanco cheese was decreased from 17.48% to 7.82% and the ratio of αS1- to αS0-casein was decreased from 1.16 to 0.43 by 10 kGy of gamma irradiation. Dose-dependent reduction of βA1-casein was also found. βA1-Casein to total protein ratios in raw milk and Queso Blanco cheese were decreased from 22.00% to 14.16% and from 21.96% to 13.89% after 10 kGy, respectively. The ratios of βA1- to βA2-casein were from 1.10 to 0.64 and 0.93 to 0.57 in milk and Queso Blanco cheese, respectively. However, αS0-, βB-, and βA3-casein increased by irradiation at 10 kGy. The results suggest that αS1-casein and βA1-casein were more susceptible to gamma irradiation, and may be related to the reduction of milk allergenicity caused by gamma irradiation

  5. Treatment of eggshell with casein phosphopeptide reduces the severity of ovariectomy-induced bone loss

    OpenAIRE

    Kim, Jung-Hoon; Kim, Min Seuk; Oh, Hong-Geun; Lee, Hak-Yong; Park, Jeong-Woo; Lee, Bong-Gun; Park, Sang-Hoon; Moon, Dae-In; Shin, Eun-Hye; Oh, Eun-Kyeong; Erkhembaatar, Munkhsoyol; Kim, Okjin; Lee, Yong-Rae; Chae, Han-Jung

    2013-01-01

    It has been generally accepted that calcium intake prevents bone loss, and frequent fracture resulted from osteoporosis. However, it is still elusive as to how effective sole calcium intake is in preventing or attenuating the severity of osteoporosis. Here, we demonstrate the effects of eggshell-casein phosphopeptide (ES-CPP), and compared these effects those of calcium supplement, for restoring ovariectomy-mediated bone loss. CPP, synthesized from the hydrolysis of casein (0.5%) using trypsi...

  6. Porphyromonas gingivalis Cysteine Proteinase Inhibition by κ-Casein Peptides ▿

    OpenAIRE

    Toh, Elena C. Y.; Dashper, Stuart G.; Huq, N. Laila; Attard, Troy J.; O'Brien-Simpson, Neil M.; Chen, Yu-Yen; Cross, Keith J.; Stanton, David P.; Paolini, Rita A.; Eric C. Reynolds

    2010-01-01

    Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. The Arg-specific (RgpA/B) and Lys-specific (Kgp) cysteine proteinases of P. gingivalis are major virulence factors for the bacterium. In this study κ-casein(109-137) was identified in a chymosin digest of casein as an inhibiting peptide of the P. gingivalis proteinases. The peptide was synthesized and shown to inhibit proteolytic activity associat...

  7. Lactogenic Hormonal Induction of Long Distance Interactions between β-Casein Gene Regulatory Elements*

    OpenAIRE

    Kabotyanski, Elena B.; Rijnkels, Monique; Freeman-Zadrowski, Courtneay; Buser, Adam C.; Edwards, Dean P.; Rosen, Jeffrey M.

    2009-01-01

    Lactogenic hormone regulation of β-casein gene expression in mammary epithelial cells provides an excellent model in which to study the mechanisms by which steroid and peptide hormone signaling control gene expression. Prolactin- and glucocorticoid-mediated induction of β-casein gene expression involves two principal regulatory regions, a proximal promoter and a distal enhancer located in the mouse approximately −6 kb upstream of the transcription start site. Using a chromosome conformation c...

  8. Colonic protein fermentation and promotion of colon carcinogenesis by thermolyzed casein.

    OpenAIRE

    Corpet, Denis; Yin, Y; X. M. Zhang; Rémésy, C.; Stamp, D; Medline, A; Thompson, L.U.; Bruce, W R; Archer, M.C.

    1995-01-01

    Thermolyzed casein is known to promote the growth of aberrant crypt foci (ACF) and colon cancer when it is fed to rats that have been initiated with azoxymethane. We speculated that the promotion was a consequence of increased colonic protein fermentation (i.e., that the thermolysis of the casein decreases its digestibility, increases the amount of protein reaching the colon, and increases colonic protein fermentation and that the potentially toxic products of this fermentation promote colon ...

  9. Colonic Protein Fermentation and Promotion of Colon Carcinogenesis by Thermolyzed Casein

    OpenAIRE

    Corpet, Denis E; Rémésy, Christian; Medline, Alan; Thompson, Lilian; Bruce, W. Robert; Archer, Michael C.

    1995-01-01

    Thermolyzed casein is known to promote the growth of aberrant crypt foci (ACF) and colon cancer when it is fed to rats that have been initiated with azoxymethane. We speculated that the promotion was a consequence of increased colonic protein fermentation (i.e., that the thermolysis of the casein decreases its digestibility, increases the amount of protein reaching the colon, and increases colonic protein fermentation and that the potentially toxic products of this fermentation promote colon ...

  10. Factors affecting antioxidant activity of soybean meal and caseine protein hydrolysates

    International Nuclear Information System (INIS)

    Antioxidative activity of protein hydrolysates was dependent on the raw material, condition of hydrolysis and lipid substrate used in model systems. Soybean meal hydrolysate was more active in lard and in linoleic acid emulsion than caseine hydrolysate, whereas caseine was more active in vegetable oils. Antioxidant activity of evaluated protein hydrolysates in all lipid systems, with or without oxidation catalysts, suggests them as natural food additives for lipid stabilization, thus for improvement of its nutritional value and sensory properties

  11. Antioxidant activity of camel milk casein before and after in vitro simulated enzymatic digestion

    OpenAIRE

    Zeineb Jrad; Jean-Michel Girardet; Isabelle Adt; Nadia Oulahal; Pascal Degraeve; Touhami Khorchani

    2014-01-01

    The effect of a successive in vitro hydrolysis by pepsin and pancreatin on the free radical scavenging activity of camel milk casein was investigated in order to assess the effect of gastro-intestinal digestion. Hydrolysis of camel casein was controlled by reversed-phase high performance liquid chromatography. Anti-oxidant activity was measured by the 2,2’-azino-bis-(3-ethylbensothiazoline-6- sulfonic acid) (ABTS) method. The Trolox equivalent antioxidant capacity (TEAC) values of camel casei...

  12. Cytotoxicity assessment of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste

    OpenAIRE

    Sandra Kalil Bussadori; Elaine Marcilio Santos; Carolina Cardoso Guedes; Lara Jansiski Motta; Kristianne Porta Santos Fernandes; Raquel Agnelli Mesquita-Ferrari; Laura Hermida Bruno; Diana Ram

    2010-01-01

    Introduction: Casein phosphopeptides (CPP) have been shown to be good carriers of calcium, phosphate, and hydroxide ions to promote enamel remineralization with applications in oral care products, professional dental products, and food products. Objectives: Evaluate the cytotoxicity of a casein phosphopeptideamorphous calcium phosphate (CPP-ACP) paste in rat fibroblasts. Materials and methods: Cytotoxicity was measured by the Trypan blue dye exclusion assay and the MTT assay. Results: Long te...

  13. NMR Water Relaxation and Self-Diffusion in Highly Concentrated Casein Systems

    OpenAIRE

    Schorr, D.; Cambert, M.; Bouchoux, A.; Gésan-Guiziou, G.; Mariette, F.

    2010-01-01

    In a variety of dairy processing operations (drying, membrane filtration...), milk is concentrated, leading to dense and ultimately solid dispersions of caseins. In this work, we explore how water mobility is affected in such conditions and if there is any relation between the properties of water as seen by NMR and the macroscopic phase behavior of the dispersions. We also investigate the effect of structure by comparing the NMR results obtained with dispersions of casein micelles (NPC, nativ...

  14. Influence of mineral environment on the buffering capacity of casein micelles

    OpenAIRE

    Salaun, Françoise; Mietton, Bernard; Gaucheron, Frederic

    2007-01-01

    The buffering capacity of casein micelle suspensions is a physico-chemical characteristic depending on the mineral and protein composition, on the interactions between these compounds and on the physicochemical environment. In this study, buffering capacity was maximum at pH about 5.2 and was mainly due to the acid-base properties of inorganic phosphate solubilised during acidification and organic phosphate from phosphoserine residues of caseins. Addition of calcium chloride to suspensions of...

  15. 54Mn absorption and excretion in rats fed soy protein and casein diets

    International Nuclear Information System (INIS)

    Rats were fed diets containing either soy protein or casein and different levels of manganese, methionine, phytic acid, or arginine for 7 days and then fed test meals labeled with 2 microCi of 54Mn after an overnight fast. Retention of 54Mn in each rat was measured every other day for 21 days using a whole-body counter. Liver manganese was higher (P less than 0.0001) in soy protein-fed rats (8.8 micrograms/g) than in casein-fed rats (5.2 micrograms/g); manganese superoxide dismutase activity also was higher in soy protein-fed rats than in casein-fed rats (P less than 0.01). There was a significant interaction between manganese and protein which affected manganese absorption and biologic half-life of 54Mn. In a second experiment, rats fed soy protein-test meals retained more 54Mn (P less than 0.001) than casein-fed rats. Liver manganese (8.3 micrograms/g) in the soy protein group was also higher than that (5.7 micrograms/g) in the casein group (P less than 0.0001), but manganese superoxide dismutase activity was unaffected by protein. Supplementation with methionine increased 54Mn retention from both soy and casein diets (P less than 0.06); activity of manganese superoxide dismutase increased (P less than 0.05) but liver manganese did not change. The addition of arginine to casein diets had little effect on manganese bioavailability. Phytic acid affected neither manganese absorption nor biologic half-life in two experiments, but it depressed liver manganese in one experiment. These results suggest that neither arginine nor phytic acid was the component in soy protein which made manganese more available from soy protein diets than casein diets

  16. Effects of 'casoparan', a peptide isolated from casein hydrolysates with mastoparan-like properties

    OpenAIRE

    Silva, Maria C. C. de Sousa e; Juliano, Maria A; Luiz Juliano; Valéria Cavallaro; Ivo Lebrun

    1992-01-01

    Casein, a protein found in milk of several species, is divided into different chains from 19 to 25 kDa. Casein is also considered as a source of amino acids and generating peptides with biological activities such as opiate, immunostimulating, antibacterial, peptidase inhibitors, among others. In this work, Sephadex G-10 chromatography followed by high-performance liquid chromatography isolation purified NZCase TT, an industrial culture media for tetanus toxin production. In the first step, fo...

  17. Translational efficiency of casein transcripts in the mammary tissue of lactating ruminants

    OpenAIRE

    Bevilacqua, Claudia; Helbling, Jean-Christophe; Miranda, Guy

    2006-01-01

    Caseins are essentially concentrated in the colloidal fraction of ruminant milks as highly hydrated and mineralized spherical particles, termed casein micelles. They form a group of four peptide chains ($\\alpha_{\\rm s1}$, $\\beta$, $\\alpha_{\\rm s2}$ and $\\kappa$), encoded by four structural genes (CSN1S1, CSN2, CSN1S2 and CSN3, respectively) of which the expression is regulated by lactogenic hormones. These phosphoproteins are synthesized, essentially during lactation, in the mammary epithelia...

  18. Tubulin phosphorylation by casein kinase II is similar to that found in vivo

    OpenAIRE

    1987-01-01

    Purified brain tubulin subjected to an exhaustive phosphatase treatment can be rephosphorylated by casein kinase II. This phosphorylation takes place mainly on a serine residue, which has been located at the carboxy- terminal domain of the beta-subunit. Interestingly, tubulin phosphorylated by casein kinase II retains its ability to polymerize in accordance with descriptions by other authors of in vivo phosphorylated tubulin. Moreover, the V8 phosphopeptide patterns of both tubulin phosphoryl...

  19. Formation of free-standing sterilized edible-films from irradiated caseinates

    International Nuclear Information System (INIS)

    γ-irradiation was used to produce free-standing sterilized edible films based on milk protein, namely sodium-caseinate and calcium-caseinate. The nature of the counter-ion as well as the protein and glycerol concentrations were examined. Irradiation of solution based on calcium-caseinate produced more crosslinks than solution based on sodium-caseinate. As a consequence, films based on calcium-caseinate showed a better mechanical strength. Glycerol was found to play a double role in enhancing the formation of crosslinks within caseinate chains, accounting for the increase of the puncture strength, and acting as a plasticizer, being responsible for the improved film extensibility and viscoelasticity. Moreover, the effect of the irradiation on the mechanical properties were strongly dependent on the glycerol/protein ratio, i.e. the formulation of the films. Films of high quality and a satisfactory mechanical behaviour were generated at glycerol/protein ratios of 0.5 and 0.67

  20. Cloning and sequencing of the gene for human β-casein

    International Nuclear Information System (INIS)

    Human β-casein is a major protein in human milk. This protein is part of the casein micelle and has been suggested to have several physiological functions in the newborn. Since there is limited information on βcasein and the factors that affect its concentration in human milk, the authors have isolated and sequenced the gene for this protein. A human mammary gland cDNA library (Clontech) in gt 11 was screened by plaque hy-hybridization using a 42-mer synthetic 32p-labelled oligo-nucleotide. Positive clones were identified and isolated, DNA was prepared and the gene isolated by cleavage with EcoR1. Following subcloning (PUC18), restriction mapping and Southern blotting, DNA for sequencing was prepared. The gene was sequenced by the dideoxy method. Human β-casein has 212 amino acids and the amino acid sequence deducted from the nucleotide sequence is to 91% identical to the published sequence for human β-casein show a high degree of conservation at the leader peptide and the highly phosphorylated sequences, but also deletions and divergence at several positions. These results provide insight into the structure of the human β-casein gene and will facilitate studies on factors affecting its expression

  1. On studying protein phosphorylation patterns using bottom-up LC-MS/MS: the case of human alpha-casein

    DEFF Research Database (Denmark)

    Kjeldsen, Frank; Savitski, Mikhail M; Nielsen, Michael L;

    2007-01-01

    occupancy rates of phosphosites in proteins may differ by orders of magnitude, and thus the occupancy rate must be reported for each occupied phosphosite. To highlight potential pitfalls in quantifying the occupancy rates, alpha(s1)-casein from human milk was selected as a model molecule representing...... moderately phosphorylated proteins. For this purpose, human milk from one Caucasian woman in the eighth month of lactation was used. The phosphorylation level of caseins is believed to have major implications for the formation of micelles that are involved in delivering valuable calcium phosphate and other...... minerals to the new-born. Human alpha(s1)-casein has been reported to be much less phosphorylated than ruminant caseins, which may indicate a different function of caseins in humans. Revealing the phosphorylation pattern in human casein can thus shed light on its function. The current study found that the...

  2. Physico-chemical changes in casein micelles of buffalo and cow milks as a function of alkalinisation

    OpenAIRE

    Ahmad, Sarfraz; Piot, Michel; Rousseau, Florence; Grongnet, Jean-François; Gaucheron, Frederic

    2009-01-01

    By modifying the forces (hydrophobic and electrostatic interactions, hydrogen bonding and presence of micellar calcium phosphate) responsible for the structure and the stability of casein micelles, alkalinisation induces a disruption of casein micelles in milk. The objective of this work was to compare the alkalinisation-induced physico-chemical changes of casein micelles of buffalo and cow milks with a special attention to the mineral fraction. The whiteness and viscosity were determined as ...

  3. Formation and properties of the whey protein/$\\kappa$-casein complexes in heated skim milk - A review

    OpenAIRE

    Donato, Laurence; Guyomarc'H, Fanny

    2009-01-01

    Abstract – The formation of complexes between whey proteins and κ-casein during heat treatment of milk dramatically affects the protein organisation in both the colloidal casein and the serum phases of milk and consequently, its technological applications. This paper reviews the composition and building interactions of these complexes and their localisation between the casein micelle and lactoserum. The currently proposed mechanisms that lead to their formation are also presented. The physico...

  4. TGF beta suppresses casein synthesis in mouse mammary explants and may play a role in controlling milk levels during pregnancy

    OpenAIRE

    1993-01-01

    Mammary explants from 14-15-d-pregnant mice synthesize and secrete milk proteins in culture in response to insulin, hydrocortisone, and prolactin. Here we demonstrate that transforming growth factor beta (TGF beta) treatment suppresses, in a dose dependent and reversible manner, the ability of explants to synthesize and secrete milk caseins. TGF beta does not affect the level of casein mRNA within explants but inhibits casein synthesis posttranscriptionally. We also show increased expression ...

  5. Assessment of Protective Effect of Amorphous Calcium Phosphate-Caseine

    Directory of Open Access Journals (Sweden)

    Anousheh Rashed-Mohassel

    2014-07-01

    Full Text Available Background: One of the complications of Iron drop recommended for 6-24 months children is the potential reduction in microhardness of primary tooth enamel because of low pH. The objective of this study is to assess the protective effect of amorphous calcium phosphate caseine phosphopeptide (ACP-CPP and silicone oil in primary teeth. Materials and Methods: Thirty extracted primary anterior teeth were divided into three equal groups. The initial micro hardness was measured by Vicker’s microhardness tester. The first group without a protective layer and the second and third group after application of ACP-CPP and silicone oil respectively, were immersed in iron drop. Microhardness was remeasured. One tooth in each group along with a tooth not exposed to iron drop were randomly chosen for SEM qualitative analysis. Analysis was performed with Repeated measures ANOVA with SPSS-18. Results: All groups exhibited significant decrease of micro hardness (p=0.001, however, no contrasting pattern was found between various groups. Conclusion: Neither ACP-CPP nor silicone oil could not provide a significant protection against micro hardness reduction after exposure to iron drop

  6. Casein/pectin nanocomplexes as potential oral delivery vehicles.

    Science.gov (United States)

    Luo, Yangchao; Pan, Kang; Zhong, Qixin

    2015-01-01

    Delivery systems prepared with natural biopolymers are of particular interests for applications in food, pharmaceutics and biomedicine. In this study, nanocomplex particles of sodium caseinate (NaCas) and pectin were fabricated and investigated as potential oral delivery vehicles. Nanocomplexes were prepared with three mass ratios of NaCas/pectin by acidification using glucono-δ-lactone and thermal treatment. NaCas/pectin at 1:1 mass ratio resulted in dispersions with the lowest turbidity and the smallest and most uniform nanocomplexes. Thermal treatment at 85 °C for 30 min facilitated the formation of stable, compact, and spherical nanocomplexes. Heating not only greatly increased the yield of nanocomplexes but also significantly improved the encapsulation capability of rutin studied as a model compound. Pectin in nanocomplexes delayed the hydrolysis of NaCas by pepsin at gastric conditions and enabled the controlled release of most rutin in simulated intestinal conditions. The nanocomplexes based on food-sourced biopolymers have promising features for oral delivery of nutrients and medicines. PMID:25800678

  7. Investigating cold gelation properties of recombined highly concentrated micellar casein concentrate and cream for use in cheese making.

    Science.gov (United States)

    Lu, Y; McMahon, D J; Vollmer, A H

    2016-07-01

    Highly concentrated micellar casein concentrate (HC-MCC), a potential ingredient for cheese making, contains ~20% casein with ~70% of serum proteins removed by microfiltration and diafiltration of skim milk, followed by vacuum evaporation. Our objective was to investigate cold gelation properties of recombined concentrated milk (RCM) by mixing thawed frozen HC-MCC and cream under different casein levels, pH, and protein-to-fat ratios, and with addition of sodium citrate or calcium. The HC-MCC was recombined with cream using low shear at 50°C for 30 min, and rheological measurements were conducted. Cold-gelling temperature [the temperature at which storage modulus (G')=loss modulus (G″)] was linearly correlated with casein levels from 8.6 to 11.5% (R(2)=0.71), pH from 6.6 to 7.0 (R(2)=0.96), and addition of sodium citrate from 0 to 0.36mmol/g of casein (R(2)=0.80). At pH 7.0, gelation occurred at 12, 26, and 38°C with 9, 10, and 11% casein, respectively. At pH 6.6, 6.8, and 7.0, RCM with 12% casein gelled at a mean temperature of 12, 26, and 37°C, respectively. Adding calcium chloride at 0.17mmol/g of casein significantly increased cold-gelling temperature from 18 to ≥50°C, whereas no significant change was observed at levels up to 0.12mmol/g of casein. Different protein to fat ratios ranging from 0.8 to 1.2 did not significantly influence gelling temperature. In transmission electron micrographs of RCM with 12% casein, casein micelles were nonspherical and partially dissociated into small protein strands. Upon addition of calcium chloride at 0.21mmol/g of casein, casein micelles were more spherical and retained colloidal structure with the presence of aggregated casein micelles. These gelation processes of RCM with or without addition of trisodium citrate were both reversible. We propose that cold gelation of RCM occurs when protein strands that have been partially released from the casein micelles entangle, restrict their mobility, and form a fine

  8. Casein Aggregates Built Step-by-Step on Charged Polyelectrolyte Film Surfaces Are Calcium Phosphate-cemented*

    OpenAIRE

    Nagy, Krisztina; Pilbat, Ana-Maria; Groma, Géza; Szalontai, Balázs; Cuisinier, Frédéric J.G.

    2010-01-01

    The possible mechanism of casein aggregation and micelle buildup was studied in a new approach by letting α-casein adsorb from low concentration (0.1 mg·ml−1) solutions onto the charged surfaces of polyelectrolyte films. It was found that α-casein could adsorb onto both positively and negatively charged surfaces. However, only when its negative phosphoseryl clusters remained free, i.e. when it adsorbed onto a negative surface, could calcium phosphate (CaP) nanoclusters bind to the casein mole...

  9. Kappa-casein gene study with molecular markers in female buffaloes (Bubalus bubalis

    Directory of Open Access Journals (Sweden)

    Antonio Roberto Otaviano

    2005-01-01

    Full Text Available Caseins comprise make up about 80% of the total protein content of milk and present polymorphism with changes in the amino acid sequence. Within this abundance of proteins, kappa-casein is noteworthy, since it has been associated with differences in milk yield, composition and processing. The objective of this study was to observe the existence of polymorphism in the kappa-casein gene in female buffaloes. For this purpose, blood samples from 115 female buffaloes, collected with vacutainer by needle punctionure of the jugular vein, were used. for genomic DNA extraction was done from blood samples. The PCR-RFLP and SSCP techniques demonstrated that the studied animals were monomorphic for the kappa-casein gene. Only allele B was observed in these animals, which was present in homozygosis. Therefore, it was not possible to quantify the gene action on milk yield and its constituents. The monomorphism observed in the population studied would allow the development of a method to identify mixtures of cow and buffalo milk in mozzarella cheese production, especially because, in cattle, the kappa-casein gene is polymorphic.

  10. The pressure-induced, lactose-dependent changes in the composition and size of casein micelles.

    Science.gov (United States)

    Wang, Pengjie; Jin, Shaoming; Guo, Huiyuan; Zhao, Liang; Ren, Fazheng

    2015-04-15

    The effects of lactose on the changes in the composition and size of casein micelles induced by high-pressure treatment and the related mechanism of action were investigated. Dispersions of ultracentrifuged casein micelle pellets with 0-10% (w/v) lactose were subjected to high pressure (400 MPa) at 20 °C for 40 min. The results indicated that the level of non-sedimentable caseins was positively related to the amount of lactose added prior to pressure treatment, and negatively correlated to the size. A mechanism for the pressure-induced, lactose-dependent changes in the casein micelles is proposed. Lactose inhibits the hydrophobic interactions between the micellar fragments during or after pressure release, through the hydrophilic layer formed by their hydrogen bonds around the micellar fragments. In addition, lactose does not favour the association between calcium and the casein aggregates after pressure release. Due to these two functions, lactose inhibited the formation of larger micelles after pressure treatment. PMID:25466047

  11. Analysis of Casein Biopolymers Adsorption to Lignocellulosic Biomass as a Potential Cellulase Stabilizer

    Directory of Open Access Journals (Sweden)

    Anahita Dehkhoda Eckard

    2012-01-01

    Full Text Available Although lignocellulosic materials have a good potential to substitute current feedstocks used for ethanol production, conversion of these materials to fermentable sugars is still not economical through enzymatic hydrolysis. High cost of cellulase has prompted research to explore techniques that can prevent from enzyme deactivation. Colloidal proteins of casein can form monolayers on hydrophobic surfaces that alleviate the de-activation of protein of interest. Scanning electron microscope (SEM, fourier transform infrared spectroscopy (FT-IR, capillary electrophoresis (CE, and Kjeldahl and BSA protein assays were used to investigate the unknown mechanism of action of induced cellulase activity during hydrolysis of casein-treated biomass. Adsorption of casein to biomass was observed with all of the analytical techniques used and varied depending on the pretreatment techniques of biomass. FT-IR analysis of amides I and II suggested that the substructure of protein from casein or skim milk were deformed at the time of contact with biomass. With no additive, the majority of one of the cellulase mono-component, 97.1 ± 1.1, was adsorbed to CS within 24 h, this adsorption was irreversible and increased by 2% after 72 h. However, biomass treatment with skim-milk and casein reduced the adsorption to 32.9% ± 6.0 and 82.8% ± 6.0, respectively.

  12. The membrane-associated form of α(s1)-casein interacts with cholesterol-rich detergent-resistant microdomains.

    Science.gov (United States)

    Le Parc, Annabelle; Honvo Houéto, Edith; Pigat, Natascha; Chat, Sophie; Leonil, Joëlle; Chanat, Eric

    2014-01-01

    Caseins, the main milk proteins, interact with colloidal calcium phosphate to form the casein micelle. The mesostructure of this supramolecular assembly markedly influences its nutritional and technological functionalities. However, its detailed molecular organization and the cellular mechanisms involved in its biogenesis have been only partially established. There is a growing body of evidence to support the concept that α(s1)-casein takes center stage in casein micelle building and transport in the secretory pathway of mammary epithelial cells. Here we have investigated the membrane-associated form of α(s1)-casein in rat mammary epithelial cells. Using metabolic labelling we show that α(s1)-casein becomes associated with membranes at the level of the endoplasmic reticulum, with no subsequent increase at the level of the Golgi apparatus. From morphological and biochemical data, it appears that caseins are in a tight relationship with membranes throughout the secretory pathway. On the other hand, we have observed that the membrane-associated form of α(s1)-casein co-purified with detergent-resistant membranes. It was poorly solubilised by Tween 20, partially insoluble in Lubrol WX, and substantially insoluble in Triton X-100. Finally, we found that cholesterol depletion results in the release of the membrane-associated form of α(s1)-casein. These experiments reveal that the insolubility of α(s1)-casein reflects its partial association with a cholesterol-rich detergent-resistant microdomain. We propose that the membrane-associated form of α(s1)-casein interacts with the lipid microdomain, or lipid raft, that forms within the membranes of the endoplasmic reticulum, for efficient forward transport and sorting in the secretory pathway of mammary epithelial cells. PMID:25549363

  13. Aggregation and structural changes of α(S1)-, β- and κ-caseins induced by homocysteinylation.

    Science.gov (United States)

    Stroylova, Yulia Y; Zimny, Jaroslaw; Yousefi, Reza; Chobert, Jean-Marc; Jakubowski, Hieronim; Muronetz, Vladimir I; Haertlé, Thomas

    2011-10-01

    Elevated homocysteine levels are resulting in N-homocysteinylation of lysyl residues in proteins and they correlate with a number of human pathologies. However, the role of homocysteinylation of lysyl residues is still poorly known. In order to study the features of homocysteinylation of intrinsically unstructured proteins (IUP) bovine caseins were used as a model. α(S1)-, β- and κ-caseins, showing different aggregations and micelle formation, were modified with homocysteine-thiolactone and their physico-chemical properties were studied. Efficiency of homocysteine incorporation was estimated to be about 1.5, 2.1 and 1.3 homocysteyl residues per one β-, α(S1)-, and κ-casein molecule, respectively. Use of intrinsic and extrinsic fluorescent markers such as Trp, thioflavin T and ANS, reveal structural changes of casein structures after homocysteinylation reflected by an increase in beta-sheet content, which in some cases may be characteristic of amyloid-like transformations. CD spectra also show an increase in beta-sheet content of homocysteinylated caseins. Casein homocysteinylation leads in all cases to aggregation. The sizes of aggregates and aggregation rates were dependent on homocysteine thiolactone concentration and temperature. DLS and microscopic studies have revealed the formation of large aggregates of about 1-3μm. Homocysteinylation of α(S1)- and β-caseins results in formation of regular spheres. Homocysteinylated κ-casein forms thin unbranched fibrils about 400-800nm long. In case of κ-casein amyloidogenic effect of homocysteinylation was confirmed by Congo red spectra. Taken together, data indicate that N-homocysteinylation provokes significant changes in properties of native caseins. A comparison of amyloidogenic transformation of 3 different casein types, belonging to the IUP protein family, shows that the efficiency of amyloidogenic transformation upon homocysteinylation depends on micellization capacity, additional disulphide bonds and

  14. The Membrane-Associated Form of αs1-Casein Interacts with Cholesterol-Rich Detergent-Resistant Microdomains

    Science.gov (United States)

    Le Parc, Annabelle; Honvo Houéto, Edith; Pigat, Natascha; Chat, Sophie; Leonil, Joëlle; Chanat, Eric

    2014-01-01

    Caseins, the main milk proteins, interact with colloidal calcium phosphate to form the casein micelle. The mesostructure of this supramolecular assembly markedly influences its nutritional and technological functionalities. However, its detailed molecular organization and the cellular mechanisms involved in its biogenesis have been only partially established. There is a growing body of evidence to support the concept that αs1-casein takes center stage in casein micelle building and transport in the secretory pathway of mammary epithelial cells. Here we have investigated the membrane-associated form of αs1-casein in rat mammary epithelial cells. Using metabolic labelling we show that αs1-casein becomes associated with membranes at the level of the endoplasmic reticulum, with no subsequent increase at the level of the Golgi apparatus. From morphological and biochemical data, it appears that caseins are in a tight relationship with membranes throughout the secretory pathway. On the other hand, we have observed that the membrane-associated form of αs1-casein co-purified with detergent-resistant membranes. It was poorly solubilised by Tween 20, partially insoluble in Lubrol WX, and substantially insoluble in Triton X-100. Finally, we found that cholesterol depletion results in the release of the membrane-associated form of αs1-casein. These experiments reveal that the insolubility of αs1-casein reflects its partial association with a cholesterol-rich detergent-resistant microdomain. We propose that the membrane-associated form of αs1-casein interacts with the lipid microdomain, or lipid raft, that forms within the membranes of the endoplasmic reticulum, for efficient forward transport and sorting in the secretory pathway of mammary epithelial cells. PMID:25549363

  15. A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression.

    OpenAIRE

    Schmidhauser, C; Casperson, G F; Myers, C A; Sanzo, K T; Bolten, S; Bissell, M.J.

    1992-01-01

    Lactogenic hormones and extracellular matrix (ECM) act synergistically to regulate beta-casein expression in culture. We have developed a functional subpopulation of the mouse mammary epithelial cell strain COMMA-1D (designated CID 9), which expresses high level of beta-casein, forms alveolar-like structures when plated onto the EHS tumor-derived matrix, and secretes beta-casein unidirectionally into a lumen. We have further shown that ECM- and prolactin-dependent regulations of beta-casein o...

  16. Bovine Chymosin: A Computational Study of Recognition and Binding of Bovine κ-Casein

    DEFF Research Database (Denmark)

    Palmer, David S.; Christensen, Anders Uhrenholt; Sørensen, Jesper; Celik, Leyla; Qvist, Karsten Bruun; Schiøtt, Hanne Birgit

    2010-01-01

    Bovine chymosin is an aspartic protease that selectively cleaves the milk protein κ-casein. The enzyme is widely used to promote milk clotting in cheese manufacturing. We have developed models of residues 97-112 of bovine κ-casein complexed with bovine chymosin, using ligand docking, conformational...... search algorithms, and molecular dynamics simulations. In agreement with limited experimental evidence, the model suggests that the substrate binds in an extended conformation with charged residues on either side of the scissile bond playing an important role in stabilizing the binding pose. Lys111 and...... Lys112 are observed to bind to the N-terminal domain of chymosin displacing a conserved water molecule. A cluster of histidine and proline residues (His98-Pro99-His100-Pro101-His102) in κ-casein binds to the C-terminal domain of the protein, where a neighboring conserved arginine residue (Arg97) is...

  17. Phase behavior of casein micelles/exocellular polysaccharide mixtures: Experiment and theory

    Science.gov (United States)

    Tuinier, R.; de Kruif, C. G.

    1999-05-01

    Dispersions of casein micelles and an exocellular polysaccharide (EPS), obtained from Lactococcus lactis subsp. cremoris NIZO B40 EPS, show a phase separation. The phase separation is of the colloidal gas-liquid type. We have determined a phase diagram that describes the separation of skim milk with EPS into a casein-micelle rich phase and an EPS rich phase. We compare the phase diagram with those calculated from theories developed by Vrij, and by Lekkerkerker and co-workers, showing that the experimental phase boundary can be predicted quite well. From dynamic light scattering measurements of the self-diffusion of the casein micelles in the presence of EPS the spinodal could be located and it corresponds with the experimental phase boundary.

  18. Chaperone-Like Activity of ß-Casein and Its Effect on Residual in Vitro Activity of Food Enzymes

    DEFF Research Database (Denmark)

    Sulewska, Anna Maria

    , tyrosinase from Agaricus bisporus and equine cytochrome c. Only for the first target β-casein was acting as a molecular chaperone i.e. its presence resulted in higher residual activity (higher degree of the function preservation). β-Casein did not have any influence on the residual activity of tyrosinase...

  19. Analysis of beta-casein gene (CSN2 polymorphism in different breeds of cattle

    Directory of Open Access Journals (Sweden)

    Martina Miluchová

    2014-11-01

    Full Text Available Normal 0 21 false false false SK X-NONE X-NONE MicrosoftInternetExplorer4 The goal of work was identification of b - casein gene polymorphism in different breeds of cow. The beta - casein constitutes up to 45 % of the casein of bovine milk. The most common forms of beta-casein in dairy cattle breeds are A1 and A2, while B is less common. The b-casein A1 variant was associated with the incidence of diabetes mellitus 1st type, coronary heart disease and autism. The A2 variant reduces serum cholesterol. The material involved 287 cows (Simmental breed – 111 cows, Pinzgau breed – 89 cows, Holstein breed – 87 cows. Bovine genomic DNA was extracted from whole blood by using commercial kit and used in order to estimate b - casein genotypes by means of PCR-RFLP method. In the populations included in the study were detected all three genotypes – homozygote genotype A1A1, heterozygote genotype A1A2 and homozygote genotype A2A2 with frequencies 0.1261, 0.3333 and 0.5405 in Simmental breed; 0.1379, 0.4598 and 0.4023 in Holstein breed, 0.3034, 0.5168 and 0.1798 in Pinzgau breed. In population of Simmental breed and Holstein breed was higher frequency of allele A2 (0.7072 and 0.6322. In opposite, in population of Pinzgau breed was present higher frequency of the allele A1 (0.5618.

  20. REVIEW OF THE CHEMISTRY OF ALPHA S-2 CASEIN AND THE GENERATION OF A HOMOLOGOUS MOLECULAR MODEL TO EXPLAIN ITS PROPERTIES

    Science.gov (United States)

    Alpha-s2-Casein comprises up to 10% of the casein fraction in bovine milk. The role of this protein in casein micelles has not been studied in detail in part due to a lack of structural information on the molecule. Interest in the role of this molecule in dairy products and nutrition has been renew...

  1. Casein SNP in Norwegian goats: additive and dominance effects on milk composition and quality

    OpenAIRE

    Lien Sigbjørn; Thaller Georg; Dagnachew Binyam S; Ådnøy Tormod

    2011-01-01

    Abstract Background The four casein proteins in goat milk are encoded by four closely linked casein loci (CSN1S1, CSN2, CSN1S2 and CSN3) within 250 kb on caprine chromosome 6. A deletion in exon 12 of CSN1S1, so far reported only in Norwegian goats, has been found at high frequency (0.73). Such a high frequency is difficult to explain because the national breeding goal selects against the variant's effect. Methods In this study, 575 goats were genotyped for 38 Single Nucleotide Polymorphisms ...

  2. Genetic Structure of Alpha S1 Casein in Slovak Pinzgau Cattle

    OpenAIRE

    Anna Trakovická; Michal Gábor

    2011-01-01

    The work was oriented to identification of -s1 casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 39 cattle. Bovine genomic DNA was isolated by commercial kit NucleoSpin Blood (Macherey-Nagel) and ethanol precipitation and used in order to estimate -s1 casein genotypes by means of PCR-RFLP method. The PCR products were digested with MaeIII restriction enzyme. In the population included in the study, homozygote genotype B...

  3. ANALYSIS OF POLYMORPHISM OF ALPHA S1 CASEIN OF SLOVAK PINZGAU CATTLE BY PCR-RFLP

    OpenAIRE

    MARTINA MILUCHOVÁ; ANNA TRAKOVICKÁ; M. GÁBOR

    2013-01-01

    The work was oriented to identification of -s1 casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 93 cattle. Bovine genomic DNA was isolated by fenol-chlorophorm deprotenization and ethanol precipitation and used in order to estimate -s1 casein genotypes by means of PCR-RFLP method. The PCR products were digested with MaeIII restriction enzyme. In the population included in the study there were homozygote genotype BB (81...

  4. Study of chemically unfolded β-casein by means of small-angle neutron scattering

    International Nuclear Information System (INIS)

    β-casein is a flexible amphiphilic milk protein which forms an unfolded conformation in presence of very high denaturant concentrations. The structure of β-casein formed at the bulk was studied by small-angle neutron scattering (SANS). The value of the second virial coefficient of the protein solutions indicates that the interactions between the polypeptide chain and solvent are repulsive. The protein conformation is similar to an excluded volume chain. The corresponding values of the contour length, L, the statistical length, b and the apparent radius of the chain cross-section, Rc are given

  5. Small-molecule inhibition of Wnt signaling through activation of casein kinase 1α

    OpenAIRE

    Thorne, Curtis A.; Hanson, Alison J.; Schneider, Judsen; Tahinci, Emilios; Orton, Darren; Cselenyi, Christopher S; Jernigan, Kristin K.; Meyers, Kelly C; Hang, Brian I.; Waterson, Alex G.; Kim, Kwangho; Melancon, Bruce; Ghidu, Victor P.; Sulikowski, Gary A.; LaFleur, Bonnie

    2010-01-01

    Wnt/β-catenin signaling is critically involved in metazoan development, stem cell maintenance and human disease. Using Xenopus laevis egg extract to screen for compounds that both stabilize Axin and promote β-catenin turnover, we identified an FDA-approved drug, pyrvinium, as a potent inhibitor of Wnt signaling (EC50 of ~10 nM). We show pyrvinium binds all casein kinase 1 (CK1) family members in vitro at low nanomolar concentrations and pyrvinium selectively potentiates casein kinase 1α (CK1α...

  6. Emulsifying properties of maillard conjugates produced from sodium caseinate and locust bean gum

    OpenAIRE

    F. A. Perrechil; Santana, R.C.; D. B. Lima; M. Z. Polastro; Cunha, R. L.

    2014-01-01

    Emulsifying properties of sodium caseinate -locust bean gum Maillard conjugates produced at different temperatures (54 - 96 ºC), protein/polysaccharide ratios (0.3 - 1.0) and reaction times (1 - 24 hours) were evaluated. Conjugate formation was confirmed by formation of color and high molecular weight fractions and the decrease of the αs- and β-casein bands. The emulsions stabilized by Maillard conjugates showed good stability. The mean droplet diameter (d32) tended to decrease with the incre...

  7. Tissue-specific expression of the rat beta-casein gene in transgenic mice.

    OpenAIRE

    Lee, K. F.; DeMayo, F J; Atiee, S H; Rosen, J. M.

    1988-01-01

    The rat beta-casein gene is a member of a small gene family, encoding the principal milk proteins. In order to understand the mechanisms by which its stage- and tissue-specific expression are regulated, initially, a 14 kb genomic clone containing the entire 7.5 kb rat beta-casein gene with 3.5 kb of 5' and 3.0 kb of 3' flanking DNA was microinjected into the germline of mice. Eight F0 transgenic mice were generated with copy numbers ranging from 1-10; five transmitted the transgene to their o...

  8. Differential regulation of rat beta-casein-chloramphenicol acetyltransferase fusion gene expression in transgenic mice.

    OpenAIRE

    Lee, K. F.; Atiee, S H; Rosen, J. M.

    1989-01-01

    Previous studies in our laboratory have demonstrated the mammary-specific expression of the entire rat beta-casein gene with 3.5 kilobases (kb) of 5' and 3.0 kb of 3' DNA in transgenic mice (Lee et al., Nucleic Acids Res. 16:1027-1041, 1988). In an attempt to localize sequences that dictate this specificity, lines of transgenic mice carrying two different rat beta-casein promoter-bacterial chloramphenicol acetyltransferase (cat) fusion genes have been established. Twenty and eight lines of tr...

  9. Study of chemically unfolded {beta}-casein by means of small-angle neutron scattering

    Energy Technology Data Exchange (ETDEWEB)

    Aschi, Adel [Laboratoire de Physique de la Matiere Molle, Faculte des Sciences de Tunis, Campus Universitaire, 1060, Tunis (Tunisia)]. E-mail: aschi13@yahoo.fr; Gharbi, Abdelhafidh [Laboratoire de Physique de la Matiere Molle, Faculte des Sciences de Tunis, Campus Universitaire, 1060, Tunis (Tunisia); Daoud, Mohamed [Service de Physique de l' Etat Condense. CEA Saclay. 91191 Gif-sur-Yvette cedex (France); Douillard, Roger [Equipe de Biochimie des Macromolecules Vegetales, Centre de Recherche Agronomique, 2Esplanade R. Garros, BP 224, 51686 Reims cedex 2 (France); Calmettes, Patrick [Laboratoire Leon Brillouin, CEA Saclay, 91191 Gif-sur-Yvette cedex (France)

    2007-01-01

    {beta}-casein is a flexible amphiphilic milk protein which forms an unfolded conformation in presence of very high denaturant concentrations. The structure of {beta}-casein formed at the bulk was studied by small-angle neutron scattering (SANS). The value of the second virial coefficient of the protein solutions indicates that the interactions between the polypeptide chain and solvent are repulsive. The protein conformation is similar to an excluded volume chain. The corresponding values of the contour length, L, the statistical length, b and the apparent radius of the chain cross-section, R{sub c} are given.

  10. Genomic organization of the bovine alpha-S1 casein gene.

    OpenAIRE

    Koczan, D; Hobom, G.; Seyfert, H.M.

    1991-01-01

    We report the sequence of the complete bovine alpha-s1 casein gene eludicating for the first time the genomic organization of an alpha-s type casein gene. Extending over 17508 bp the gene is split into 19 exons, ranging in size from 24 bp to 385 bp. Except for the translational stop codon not a single coding triplet of the alpha-s1 reading frame is disrupted by any of the splice junctions, which all confirm to known splice consensus sequences. Nine out of 16 coding exons begin with a 'GAX' co...

  11. Temperature-dependent dynamics of bovine casein micelles in the range 10-40 °C.

    Science.gov (United States)

    Liu, Dylan Z; Weeks, Michael G; Dunstan, David E; Martin, Gregory J O

    2013-12-15

    Milk is a complex colloidal system that responds to changes in temperature imposed during processing. Whilst much has been learned about the effects of temperature on milk, little is known about the dynamic response of casein micelles to changes in temperature. In this study, a comprehensive physico-chemical study of casein micelles in skim milk was performed between 10 and 40 °C. When fully equilibrated, the amount of soluble casein, soluble calcium and the pH of skim milk all decreased as a function of increasing temperature, whilst the hydration and volume fraction of the casein micelles decreased. The effect of temperature on casein micelle size, as determined by dynamic light scattering and differential centrifugation, was less straightforward. Real-time measurements of turbidity and pH were used to investigate the dynamics of the system during warming and cooling of milk in the range 10-40 °C. Changes in pH are indicative of changes to the mineral system and the turbidity is a measure of alterations to the casein micelles. The pH and turbidity showed that alterations to both the casein micelles and the mineral system occurred very rapidly on warming. However, whilst mineral re-equilibration occurred very rapidly on cooling, changes to the casein micelle structure continued after 40 min of measurement, returning to equilibrium after 16 h equilibration. Casein micelle structure and the mineral system of milk were both dependent on temperature in the range 10-40 °C. The dynamic response of the mineral system to changes in temperature appeared almost instantaneous whereas equilibration of casein was considerably slower, particularly upon cooling. PMID:23993588

  12. Casein infusion rate influences feed intake differently depending on metabolizable protein balance in dairy cows: A multilevel meta-analysis.

    Science.gov (United States)

    Martineau, R; Ouellet, D R; Kebreab, E; Lapierre, H

    2016-04-01

    The effects of casein infusion have been investigated extensively in ruminant species. Its effect on responses in dry matter intake (DMI) has been reviewed and indicated no significant effect. The literature reviewed in the current meta-analysis is more extensive and limited to dairy cows fed ad libitum. A total of 51 studies were included in the meta-analysis and data were fitted to a multilevel model adjusting for the correlated nature of some studies. The effect size was the mean difference calculated by subtracting the means for the control from the casein-infused group. Overall, casein infusion [average of 333g of dry matter (DM)/d; range: 91 to 1,092g of DM/d] tended to increase responses in DMI by 0.18kg/d (n=48 studies; 3 outliers). However, an interaction was observed between the casein infusion rate (IR) and the initial metabolizable protein (MP) balance [i.e., supply minus requirements (NRC, 2001)]. When control cows were in negative MP balance (n=27 studies), responses in DMI averaged 0.28kg/d at mean MP balance (-264g/d) and casein IR (336g/d), and a 100g/d increment in the casein IR from its mean increased further responses by 0.14kg/d (MP balance being constant), compared with cows not infused with casein. In contrast, when control cows were in positive MP balance (n=22 studies; 2 outliers), responses in DMI averaged -0.20kg/d at mean casein IR (339g/d), and a 100g/d increment in the casein IR from its mean further decreased responses by 0.33kg/d, compared with cows not infused with casein. Responses in milk true protein yield at mean casein IR were greater (109 vs. 65g/d) for cows in negative vs. positive MP balance, respectively, and the influence of the casein IR on responses was significant only for cows in negative MP balance. A 100g/d increment in the casein IR from its mean increased further responses in milk true protein yield by 25g/d, compared with cows not infused with casein. Responses in blood urea concentration increased in casein

  13. Whey and casein labeled with L-[1-13C]leucine and muscle protein synthesis

    DEFF Research Database (Denmark)

    Reitelseder, Søren; Agergaard, Jakob; Doessing, Simon;

    2011-01-01

    mass), or a noncaloric control drink was ingested immediately after exercise. l-[1-(13)C]leucine-labeled whey and casein were used while muscle protein synthesis (MPS) was assessed. Blood and muscle tissue samples were collected to measure systemic hormone and amino acid concentrations, tracer...

  14. Development of biodegradable foamlike materials based on casein and sodium montmorillonite clay

    Science.gov (United States)

    Biodegradable foamlike materials based on a naturally occurring polymer (casein protein) and sodium montmorillonite clay (Na+-MMT) were produced through a simple freeze-drying process. By utilizing DL-glyceraldehyde (GC) as a chemical cross-linking agent, the structural integrity of these new aeroge...

  15. Molecular cloning and expression of bovine kappa-casein in Escherichia coli

    International Nuclear Information System (INIS)

    A cDNA library was constructed using poly(A)+RNA from bovine mammary gland. This cDNA library of 6000 clones was screened employing colony hybridization using 32P-labelled oligonucleotide probes and restriction endonuclease mapping. The cDNA from the selected plasmid, pKR76, was sequenced using the dideoxy-chain termination method. The cDNA insert of pKR76 carries the full-length sequence, which codes for mature kappa-casein protein. The amino acid sequence deduced from the cDNA sequence fits the published amino acid sequence with three exceptions; the reported pyroglutamic acid at position 1, tyrosine at position 35, and aspartic acid at position 81 are, respectively, a glutamine, a histidine, and an asparagine in the clone containing pKR76. The MspI-, NlaIV-cleaved fragment (630 base pair) from the kappa-casein cDNA insert has been subcloned into expression vectors pUC18 and pKK233-2, which contain a lac promoter and a trc promoter, respectively. Escherichia coli cells carrying the recombinant expression plasmids were shown to produce kappa-casein protein having the expected mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and being recognized by specific antibodies raised against natural bovine kappa-casein

  16. Trichostatin A inhibits beta-casein expression in mammary epithelial cells

    International Nuclear Information System (INIS)

    Many aspects of cellular behavior are affected by information derived from association of the extracellular matrix (ECM) and with cell membrane receptors. When cultured in the presence of laminin-containing ECM and prolactin (Prl), normal mammary epithelial cells express the milk protein beta-casein. Previously, we defined the minimal ECM- and Prl-responsive enhancer element BCE-1 from the upstream region of the beta-casein gene. We also found that BCE-1 was only active when stably integrated into chromatin, and that trichostatin A (TSA), a reagent that leads to alterations in chromatin structure, was able to activate the integrated enhancer element. We now show that endogenous b-casein gene, which is controlled by a genetic assembly that is highly similar to that of BCE-1 and which is also activated by incubation in ECM and Prl, is instead inhibited by TSA. We provide evidence that the differing response of b-casein and BCE-1 to TSA is neither due to an unusual effect of TSA on mammary epithelial cells, nor to secondary consequences from the expression of a separate gene, nor to a particular property of the BCE-1 construct. As a component of this investigation, we also showed that ECM could mediate rapid histone deacetylation in mammary epithelial cells. These results are discussed in combination with previous work showing that TSA mediates the differentiation of many types of cancer cells but inhibits differentiation of some nonmalignant cell types

  17. Phosphorylation of acidic ribosomal proteins from rabbit reticulocytes by a ribosome-associated casein kinase

    DEFF Research Database (Denmark)

    Issinger, O G

    1977-01-01

    Two acidic proteins from 80-S ribosomes were isolated and purified to homogeneity. The purified acidic proteins could be phosphorylated by casein kinase using [gamma-32P]ATP and [gamma-32P]GTP as a phosphoryl donor. The proteins became phosphorylated in situ, too. Sodium dodecyl sulfate...

  18. Hierarchical Networks of Casein Proteins: An Elasticity Study Based on Atomic Force Microscopy

    NARCIS (Netherlands)

    Uricanu, V.I.; Duits, M.H.G.; Mellema, J.

    2004-01-01

    2D- and 3D-atomic force microscopy (AFM) experiments were performed on single casein micelles (CM) in native state, submerged in liquid, using a home-built AFM instrument. The micelles were immobilized via carbodiimide chemistry to a self-assembled monolayer supported on gold-coated slides. Off-line

  19. Selective Inhibition of Casein Kinase 1 epsilon Minimally Alters Circadian Clock Period

    Czech Academy of Sciences Publication Activity Database

    Walton, K. M.; Fisher, K.; Rubitski, D.; Marconi, M.; Meng, Q.-J.; Sládek, Martin; Adams, J.; Bass, M.; Chandrasekaran, R.; Butler, T.; Griffor, M.; Rajamohan, F.; Serpa, M.; Chen, Y.; Claffey, M.; Hastings, M.; Loudon, A.; Maywood, E.; Ohren, J.; Doran, A.; Wager, T. T.

    2009-01-01

    Roč. 330, č. 2 (2009), s. 430-439. ISSN 0022-3565 Institutional research plan: CEZ:AV0Z50110509 Keywords : circadian clock * casein kinase 1 epsilon * inhibitor Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.093, year: 2009

  20. Reconstitution of normal and hyperactivated forms of casein kinase-2 by variably mutated beta-subunits

    DEFF Research Database (Denmark)

    Boldyreff, B; Meggio, F; Pinna, L A;

    1993-01-01

    Twenty-one mutants of the noncatalytic beta-subunit of human casein kinase-2 have been created, expressed in Escherichia coli, and purified to homogeneity. They are either modified at the autophosphorylation site (mutants beta delta 1-4 and beta A 5,6) or bear variable deletions in their C...

  1. Caseins from bovine colostrum and milk strongly bind piscidin-1, an antimicrobial peptide from fish.

    Science.gov (United States)

    Kütt, Mary-Liis; Stagsted, Jan

    2014-09-01

    A model system of bovine colostrum and piscidin, a fish-derived antimicrobial peptide, was developed to study potential interactions of antimicrobial peptides in colostrum. We did not detect any antimicrobial activity of colostrum using the radial plate diffusion assay; in fact colostrum completely abrogated activity of added piscidin. This could not be explained by degradation of piscidin by colostrum, which was less than ten percent. We found that colostrum even protected piscidin against degradation by added proteases. We further observed that colostrum and milk rapidly quenched the fluorescence of fluorescein-piscidin but not that of fluorescein. This effect was not seen with BSA and the specific quenching of fluorescein-piscidin by colostrum was saturably inhibited with unlabeled piscidin. Size exclusion chromatography indicated that fluorescein-piscidin bound to casein micelles with no apparent binding to IgG or whey proteins. Further, addition of pure caseins was able to quench fluorescence of fluorescein-piscidin and to inhibit the antimicrobial activity of piscidin. The interaction between caseins and piscidin could be dissociated by guanidine hydrochloride and recovered piscidin had antimicrobial activity against bacteria. Based on our results we propose that caseins could be carriers for antimicrobial peptides in colostrum and milk. PMID:25036607

  2. The effects of ginsenosides to amyloid fibril formation by RCMκ-casein.

    Science.gov (United States)

    Liu, Jihua; Chen, Fanbo; Yin, Jianyuan; Bu, Fengquan; Zheng, Baohua; Yang, Miao; Wang, Yunhua; Sun, Dandan; Meng, Qin

    2015-08-01

    When not incorporated into the casein micelle, isolated κ-casein spontaneously forms amyloid fibrils under physiological conditions, and is a convenient model for researching generic aspects of fibril formation. Ginsenosides have recently attracted much research interest because of the effects on aging diseases, which are always associated with amyloid fibril formation, for example, Alzheimer's, Parkinson's, and Huntington's diseases. In addition, the mechanism remains unclear that ginsenosides exert the effects against aging diseases. To address these aspects, we have investigated the ability of ginsenoside Rb1, Rc, Rg1, and Re influencing fibril formation by RCMκ-casein (reduced and carboxymethylated κ-casein), with the methods of Thioflavin T fluorescence assay, transmission electron microscopy (TEM), and intrinsic fluorescence spectroscopy. The results showed that ginsenoside Rb1 and Rg1 inhibited obviously RCMκ-CN fibrillation in both the initial rate and final level of ThT fluorescence. On the contrary, ginsenoside Re had a few effect on promoting RCMκ-CN fibril formation, proved by thick and larger fibrils observed frequently in TEM. While Rc did not influence RCMκ-CN fibrillation. It is demonstrated that Rg1 prevent RCMκ-CN fibril formation by stabilising RCMκ-CN in its native like state. Additional chemical structure difference of ginsenosides and the effects on fibril formation are also implicated. PMID:25934110

  3. Spectroscopic investigation of the influence of calcium ion on the structures of casein micelles.

    Science.gov (United States)

    Wang, Peng-Jie; Wu, Jian-Ping; Zhang, Hao; Guo, Hui-Yuan; Liu, Hong-Na; Ren, Fa-Zheng

    2014-01-01

    The effects of calcium ion on the structural properties of casein micelles in the course of heat treatment were synthetically examined by non-structure-invasive spectrometry. The hydrophobicity, reflected by extrinsic fluorescence (ANS fluorescence), was positively correlated with the concentration of the calcium ion, within the range of 0 to 12 mmol x L(-1). Meanwhile, the turbidity and stability of casein micelles also increased with the growth of calcium concentrations. However, opposite results were observed for hydrodynamic diameter and polydispersity index. Compared with the calcium ion, the calcium-chelator (citrate) has an opposite effect on the structural characteristics of casein micelles. Within the calcium concentrations range of 0 to 12 mmol x L(-1), the hydrophobicity, stability and turbidity were negatively correlated with the concentration of the calcium ion, nevertheless, opposite results were observed for hydrodynamic diameter and polydispersity index. All the results indicate that the calcium ion could be used to modify the structures of casein micelles during heat heatment. PMID:24783540

  4. EFFECT OF PRESSURIZED CARBON DIOXIDE (CO2) ON THE STRUCTURE OF CASEIN MICELLES

    Science.gov (United States)

    At the heart of the skim milk system are the colloidal casein–calcium–transport complexes termed the casein micelles. The application of physical chemical techniques such as light, neutron, and X-ray scattering and electron microscopy, has yielded a wealth of experimental data concerning the structu...

  5. The Gluten-Free, Casein-Free Diet and Autism: Limited Return on Family Investment

    Science.gov (United States)

    Hurwitz, Sarah

    2013-01-01

    The gluten-free, casein-free (GFCF) diet is widely used by families of children with autism spectrum disorders (ASD). Despite its popularity, there is limited evidence in support of the diet. The purpose of this article was to identify and evaluate well-controlled studies of the GFCF diet that have been implemented with children with ASD. A review…

  6. Trichostatin A inhibits beta-casein expression in mammary epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Pujuguet, Philippe; Radisky, Derek; Levy, Dinah; Lacza, Charlemagne; Bissell, Mina J.

    2002-02-22

    Many aspects of cellular behavior are affected by information derived from association of the extracellular matrix (ECM) and with cell membrane receptors. When cultured in the presence of laminin-containing ECM and prolactin (Prl), normal mammary epithelial cells express the milk protein beta-casein. Previously, we defined the minimal ECM- and Prl-responsive enhancer element BCE-1 from the upstream region of the beta-casein gene. We also found that BCE-1 was only active when stably integrated into chromatin, and that trichostatin A (TSA), a reagent that leads to alterations in chromatin structure, was able to activate the integrated enhancer element. We now show that endogenous b-casein gene, which is controlled by a genetic assembly that is highly similar to that of BCE-1 and which is also activated by incubation in ECM and Prl, is instead inhibited by TSA. We provide evidence that the differing response of b-casein and BCE-1 to TSA is neither due to an unusual effect of TSA on mammary epithelial cells, nor to secondary consequences from the expression of a separate gene, nor to a particular property of the BCE-1 construct. As a component of this investigation, we also showed that ECM could mediate rapid histone deacetylation in mammary epithelial cells. These results are discussed in combination with previous work showing that TSA mediates the differentiation of many types of cancer cells but inhibits differentiation of some nonmalignant cell types.

  7. Cloning and sequencing of the casein kinase 2 alpha subunit from Zea mays

    DEFF Research Database (Denmark)

    Dobrowolska, G; Boldyreff, B; Issinger, O G

    1991-01-01

    The nucleotide sequence of the cDNA coding for the alpha subunit of casein kinase 2 of Zea mays has been determined. The cDNA clone contains an open reading frame of 996 nucleotides encoding a polypeptide comprising 332 amino acids. The primary amino acid sequence exhibits 75% identity to the alpha...

  8. Xenopus paraxial protocadherin inhibits Wnt/β-catenin signalling via casein kinase 2β

    OpenAIRE

    Kietzmann, Anja; Wang, Yingqun; Weber, Dominik; Steinbeisser, Herbert

    2011-01-01

    Paraxial protocadherin (PAPC) is a known cell adhesion molecule that also activates the Wnt/PCP pathway. Steinbeisser and colleagues show here that PAPC not only activates the non-canonical Wnt/PCP pathway but, at the same time, also inhibits the canonical Wnt/beta-catenin pathway through interaction with casein kinase 2.

  9. Acute intestinal injury induced by acetic acid and casein: prevention by intraluminal misoprostol

    International Nuclear Information System (INIS)

    Acute injury was established in anesthetized rabbits by intraluminal administration of acetic acid with and without bovine casein, into loops of distal small intestine. Damage was quantified after 45 minutes by the blood-to-lumen movement of 51Cr-labeled ethylenediaminetetraacetic acid (EDTA) and fluorescein isothiocyanate-tagged bovine serum albumin as well as luminal fluid histamine levels. The amount of titratable acetic acid used to lower the pH of the treatment solutions to pH 4.0 was increased by the addition of calcium gluconate. Luminal acetic acid caused a 19-fold increase in 51Cr-EDTA accumulation over saline controls; casein did not modify this effect. In saline controls, loop fluid histamine levels bordered on the limits of detection (1 ng/g) but were elevated 19-fold by acetic acid exposure and markedly increased (118-fold) by the combination of acid and casein. Intraluminal misoprostol (3 or 30 micrograms/mL), administered 30 minutes before acetic acid, significantly attenuated the increase in epithelial permeability (luminal 51Cr-EDTA, fluorescein isothiocyanate-bovine serum albumin accumulation) and histamine release (P less than 0.05). Diphenhydramine, alone or in combination with cimetidine, and indomethacin (5 mg/kg IV) were not protective. It is concluded that exposure of the epithelium to acetic acid promotes the transepithelial movement of casein leading to enhanced mast cell activation and mucosal injury. Damage to the epithelial barrier can be prevented by misoprostol

  10. Impact of Casein and Gluten Free Dietary Intervention on Selected Autistic Children

    Directory of Open Access Journals (Sweden)

    Veerappan Nishadevi

    2008-09-01

    Full Text Available Objective: Autism is a life long developmental disorder that emerges in early childhood and results in significant lifelong disability. The goal of treatment is to promote the childs social and language development and minimize behaviors that interfere with the childs functioning and learning. This study evaluated the impact of casein and gluten free diet among selected autistic children. Methods: Three private special schools in Salem District, Tamilnadu, India were selected. A total number of 50 autistic children 10 from SIMEC, 10 from MMIC and 30 from CSI comprised the study sample. Background information, clinical history and nutritional status, dietary pattern were collected from the 50 selected autistic children. Out of 50 autistic children 30 autistic children were selected for the dietary intervention. Diet counseling regarding casein free diet was imparted to Group I (n=10, gluten free diet to Group II (n=10 and both casein and gluten free diet for Group III (n=10. The diet was followed for a period of 2 months. The efficacy of the dietary exclusion of casein and gluten was evaluated using a food and behavior diary on a day to day basis, using observation method. Findings: Results about Group I autistic children who followed dietary exclusion of casein free diet showed that the mean scores before and after casein free dietary intervention depiticted these improvements as, 1 to 1.2 for attention, 2.8 to 2.9 for sleep, 1.1 to 1.3 for hyperactivity, 1.1 to 1.2 for anxiety/compulsion. For Group II autistic children who followed dietary exclusion of gluten free diet showed the improvements as 1.1 to 1.4 for attention 2.5 to 3 for sleep, 1.7 to 1.9 for hyperactivity, 1.1 to 1.2 for anxiety/compulsion. About Group III autistic children who followed dietary exclusion of both casein and gluten free diet showed the improvements as 1.1 to 1.3 for attention, 2.5 to 2.7 for sleep, 1.3 to 1.7 for hyperactivity, and 1.1 to 1.2 for anxiety

  11. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells

    International Nuclear Information System (INIS)

    When primary mouse mammary epithelial cells are cultured on plastic, they rapidly lose their ability to synthesize and secrete most milk proteins even in the presence of lactogenic hormones, whereas cells cultured on release type I collagen gels show greatly enhanced mRNA levels and secretion rates of β-casein and of some other milk proteins. The authors show here that culture on a reconstituted basement membrane from Engelbreth-Holm-Swarm tumor (EHS) allows > 90% of cells to produce high levels of β-casein. By comparison, 30-40% of cells on released type 1 gels and only 2-10% of cells on plastic express β-casein after 6 days in culture. Because only 40% of cells from late pregnant gland produced β-casein before culture, the EHS matrix can both induce and maintain an increased level of casein gene expression. Individual basal lamina components were also evaluated. Type IV collagen and fibronectin had little effect on morphology and β-casein mRNA levels. In contrast, both laminin and heparan sulfate proteoglycan increased β-casein mRNA levels. Profound morphological differences were evident between cells cultured on plastic and on EHS matrix, the latter cells forming ducts, ductules, and lumina and resembling secretory alveoli. These results emphasize the vital role of the extracellular matrix in receiving and integrating structural and functional signals that can direct specific gene expression in differentiated tissues

  12. Comparison of Casein Micelles Micrographs in Raw and Pasteurized Skim Milk in Different pHs by SEM and TEM

    Directory of Open Access Journals (Sweden)

    H. Lamea

    2005-07-01

    Full Text Available In this research some properties of the casein micelles in the raw and pasteurized milk were studied by electron microscopy. SEM and TEM were used to evaluate the differences in acidified casein micelles of raw and pasteurized milk at the Iso Electric Point (pH=4.6. Milk samples were taken from research pilot plant of The College of Agriculture. Milk was pasteurized by the L.T.L.T. method in the same pilot plant. The samples of raw and pasteurized milk were divided into two parts. One part of raw and pasteurized milk was acidified to the Iso Electric Point of caseins (pH=4.6 by lactic acid (9% and then sample preparation for electron microscopy was done. According to the previous findings, results indicated that in the native pH, specially in fresh raw milk casein micelles were in spherical and individual form with the smooth surface. Aggregated casein micelles were present of acidified samples of the raw and pasteurized milk. Aggregation was the result of neutralization of electric charges in the isoelectric pH of casein and partial removal of micellar calcium phosphate. Results of both electron microscope confirmed each other and effects of heating on increasing of the casein micelle size during pasteurization were seen.

  13. Incorporation of 32P-Na2HPO4 into caseins and lipoprotein complexes of milk from a lactating buffalo

    International Nuclear Information System (INIS)

    32P-phosphate was given intravenously to a lactating buffalo to study the utilization of blood inorganic-phosphate by the mammary gland. The distribution of radioactivity in the mammary-cell-plasma membrane (MCPM), milk-fat-globule membrane (MFGM), low density lipoproteins (LDLP) and caseins was studied. Maximum radioactivity was incorporated into casein. Among the caseins, α-casein complex had minimum radioactivity. Radioactivity from γ-casein complex appeared to be incorporated into β-casein. On intravenous injection the peak specific activity of 32P appeared at the same time in the MCPM and MFGM, but on prolonged infusion it appeared first in the MCPM and then in MFGM, indicating MFGM and MCPM to be of common origin. The peak specific activity of lipids from casein micelles appeared later than that in MCPM and MFGM. Autoradiography of MCPM and MFGM lipids showed that 32P-phosphate was first incorporated into phosphatidyl choline (PC) and then into other phospholipids (PL). In LDLP, 32P-phosphate was incorporated into PC alone even after 25 hr of injection. The role of PC in PL biosynthesis in mammary gland has been discussed. (author)

  14. Identification of protein tyrosine phosphatase 1B and casein as substrates for 124-v-Mos

    Directory of Open Access Journals (Sweden)

    Stabel Silvia

    2002-04-01

    Full Text Available Abstract Background The mos proto-oncogene encodes a cytoplasmic serine/threonine-specific protein kinase with crucial function during meiotic cell division in vertebrates. Based on oncogenic amino acid substitutions the viral derivative, 124-v-Mos, displays constitutive protein kinase activity and functions independent of unknown upstream effectors of mos protein kinase. We have utilized this property of 124-v-Mos and screened for novel mos substrates in immunocomplex kinase assays in vitro. Results We generated recombinant 124-v-Mos using the baculovirus expression system in Spodoptera frugiperda cells and demonstrated constitutive kinase activity by the ability of 124-v-Mos to auto-phosphorylate and to phosphorylate vimentin, a known substrate of c-Mos. Using this approach we analyzed a panel of acidic and basic substrates in immunocomplex protein kinase assays and identified novel in vitro substrates for 124-v-Mos, the protein tyrosine phosphatase 1B (PTP1B, alpha-casein and beta-casein. We controlled mos-specific phosphorylation of PTP1B and casein in comparative assays using a synthetic kinase-inactive 124-v-Mos mutant and further, tryptic digests of mos-phosphorylated beta-casein identified a phosphopeptide specifically targeted by wild-type 124-v-Mos. Two-dimensional phosphoamino acid analyses showed that 124-v-mos targets serine and threonine residues for phosphorylation in casein at a 1:1 ratio but auto-phosphorylation occurs predominantly on serine residues. Conclusion The mos substrates identified in this study represent a basis to approach the identification of the mos-consensus phosphorylation motif, important for the development of specific inhibitors of the Mos protein kinase.

  15. Purification and characterization of a casein kinase 2-type protein kinase from pea nuclei

    Science.gov (United States)

    Li, H.; Roux, S. J.

    1992-01-01

    Almost all the polyamine-stimulated protein kinase activity associated with the chromatin fraction of nuclei purified from etiolated pea (Pisum sativum L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.35 molar NaCl. This protein kinase can be further purified over 2000-fold by salt fractionation and anion-exchange and casein-agarose column chromatography, after which it is more than 90% pure. The purified kinase has a specific activity of about 650 nanomoles per minute per milligram protein in the absence of polyamines, with either ATP or GTP as phosphoryl donor. Spermidine can stimulate its activity fourfold, with half-maximal activation at about 2 millimolar. Spermine and putrescine also stimulate activity, although somewhat less effectively. This kinase has a tetrameric alpha 2 beta 2 structure with a native molecular weight of 130,000, and subunit molecular weights of 36,000 for the catalytic subunit (alpha) and 29,000 for the regulatory subunit (beta). In western blot analyses, only the alpha subunit reacts strongly with polyclonal antibodies to a Drosophila casein kinase II. The pea kinase can use casein and phosvitin as artificial substrates, phosphorylating both the serine and threonine residues of casein. It has a pH optimum near 8.0, a Vmax of 1.5 micromoles per minute per milligram protein, and a Km for ATP of approximately 75 micromolar. Its activity can be almost completely inhibited by heparin at 5 micrograms per milliliter, but is relatively insensitive to concentrations of staurosporine, K252a, and chlorpromazine that strongly antagonize Ca(2+) -regulated protein kinases. These results are discussed in relation to recent findings that casein kinase 2-type kinases may phosphorylate trans-acting factors that bind to light-regulated promoters in plants.

  16. Water interactions with varying molecular states of bovine casein: 2H NMR relaxation studies

    International Nuclear Information System (INIS)

    The caseins occur in milk as spherical colloidal complexes of protein and salts with an average diameter of 1200 A, the casein micelles. Removal of Ca2+ is thought to result in their dissociation into smaller protein complexes stabilized by hydrophobic interactions and called submicelles. Whether these submicelles actually occur within the micelles as discrete particles interconnected by calcium phosphate salt bridges has been the subject of much controversy. A variety of physical measurements have shown that casein micelles contain an inordinately high amount of trapped water (2 to 7 g H2O/g protein). With this in mind it was of interest to determine if NMR relaxation measurements could detect the presence of this trapped water within the micelles, and to evaluate whether it is a continuum with picosecond correlation times or is associated in part with discrete submicellar structures with nanosecond motions. For this purpose the variations in 2H NMR longitudinal and transverse relaxation rates of water with protein concentration were determined for bovine casein at various temperatures, under both submicellar and micellar conditions. D2O was used instead of H2O to eliminate cross-relaxation effects. From the protein concentration dependence of the relaxation rates, the second virial coefficient of the protein was obtained by nonlinear regression analysis. Using either an isotropic tumbling or an intermediate asymmetry model, degrees of hydration, v, and correlation times, tau c, were calculated for the caseins; from the latter parameter the Stokes radius, r, was obtained. Next, estimates of molecular weights were obtained from r and the partial specific volume. Values were in the range of those published from other methodologies for the submicelles

  17. Structural heterogeneity of milk casein micelles: a SANS contrast variation study.

    Science.gov (United States)

    Bouchoux, Antoine; Ventureira, Jorge; Gésan-Guiziou, Geneviève; Garnier-Lambrouin, Fabienne; Qu, Peng; Pasquier, Coralie; Pézennec, Stéphane; Schweins, Ralf; Cabane, Bernard

    2015-01-14

    We examine the internal structure of milk casein micelles using the contrast variation method in Small-Angle Neutron Scattering (SANS). Experiments were performed with casein dispersions of different origins (i.e., milk powder or fresh milk) and extended to very low q-values (∼9 × 10(-4) Å(-1)), thus making it possible to precisely determine the apparent gyration radius Rg at each contrast. From the variation of I(q → 0) with contrast, we determine the distribution of composition of all the particles in the dispersions. As expected, most of these particles are micelles, made of casein and calcium phosphate, with a narrow distribution in compositions. These micelles always coexist with a very small fraction of fat droplets, with sizes in the range of 20-400 nm. For the dispersions prepared from fresh milk, which were purified under particularly stringent conditions, the number ratio of fat droplets to casein micelles is as low as 1 to 10(6). In that case, we are able to subtract from the total intensity the contribution of the fat droplets and in this way obtain the contribution of the micelles only. We then analyze the variation of this contribution with contrast using the approach pioneered by H. B. Stuhrmann. We model the casein micelle as a core-shell spherical object, in which the local scattering length density is determined by the ratio of calcium phosphate nanoclusters to proteins. We find that models in which the shell has a lower concentration of calcium phosphate than the core give a better agreement than models in which the shell has a higher density than the core. PMID:25388767

  18. Effect of Casein Concentration in Suspensions and Gels on Poly(ethylene glycol)s NMR Self-Diffusion Measurements

    OpenAIRE

    Colsenet, R.; Söderman, O.; Mariette, F.

    2005-01-01

    PFG-NMR spectroscopy was used to study the diffusion of molecular probes (poly(ethyleneglycol)s) in casein suspensions and gels in terms of the effects of probe molecular size (molecular mass between 1080 and 634 000 g/mol), casein concentrations (from 3.24 to 16.22 g/100 g), and effects of rennet coagulation. A strong dependency of diffusion on probe size was observed, both in casein suspensions and in gels: as the PEG size increased, the diffusion was reduced. This effect was more pronou...

  19. The extracellular PI-type proteinase of Lactococcus lactis hydrolyzes beta-casein into more than one hundred different oligopeptides.

    OpenAIRE

    Juillard, V; van der Laan, H.; Kunji, E R; Jeronimus-Stratingh, C M; Bruins, A P; Konings, W. N.

    1995-01-01

    The peptides released from beta-casein by the action of PI-type proteinase (PrtP) from Lactococcus lactis subsp. cremoris Wg2 have been identified by on-line coupling of liquid chromatography to mass spectrometry. After 24 h of incubation of beta-casein with purified PrtP, a stable mixture of peptides was obtained. The trifluoroacetic acid-soluble peptides of this beta-casein hydrolysate were fractionated by high-performance liquid chromatography and introduced into the liquid chromatography-...

  20. Molecular cloning of casein kinase II alpha subunit from Dictyostelium discoideum and its expression in the life cycle.

    OpenAIRE

    Kikkawa, U; Mann, S K; Firtel, R A; Hunter, T

    1992-01-01

    A Dictyostelium discoideum cDNA encoding an alpha-type subunit of casein kinase II was isolated, and its cDNA was used to study developmental expression of casein kinase II during the Dictyostelium life cycle. The 1.3-kb cDNA insert contained an open reading frame of 337 amino acids (M(r) 39,900). The deduced amino acid sequence has high homology with those of casein kinase II alpha subunits from other species. Genomic Southern blot analysis suggested that there is a single gene encoding case...

  1. Genetic polymorphisms of αS1-, αS2- and k-casein in Maltese goat breed

    OpenAIRE

    A. Caroli; G. Erhardt; C. Dario; G. Ceriotti; Chessa, S.

    2011-01-01

    The casein genes are organized as a tight cluster including αS1- (CSN1S1), β- (CSN2), αS2- (CSN1S2) and k-casein (CSN3). Investigation at casein haplotype level is therefore necessary to detect effects which could be used for the genetic improvement of goat breeds aiming to preserve biodiversity, safeguard typical products, and valorise nutritional properties of goat milk. Mutations responsible for the reduced or null expression of CSN1S1........

  2. Genetic polymorphisms of αS1-, αS2- and k-casein in Maltese goat breed

    Directory of Open Access Journals (Sweden)

    A. Caroli

    2011-03-01

    Full Text Available The casein genes are organized as a tight cluster including αS1- (CSN1S1, β- (CSN2, αS2- (CSN1S2 and k-casein (CSN3. Investigation at casein haplotype level is therefore necessary to detect effects which could be used for the genetic improvement of goat breeds aiming to preserve biodiversity, safeguard typical products, and valorise nutritional properties of goat milk. Mutations responsible for the reduced or null expression of CSN1S1........

  3. Casein kinase I-like protein kinases encoded by YCK1 and YCK2 are required for yeast morphogenesis.

    OpenAIRE

    Robinson, L. C.; Menold, M. M.; Garrett, S.; Culbertson, M R

    1993-01-01

    Casein kinase I is an acidotropic protein kinase class that is widely distributed among eukaryotic cell types. In the yeast Saccharomyces cerevisiae, the casein kinase I isoform encoded by the gene pair YCK1 and YCK2 is a 60- to 62-kDa membrane-associated form. The Yck proteins perform functions essential for growth and division; either alone supports growth, but loss of function of both is lethal. We report here that casein kinase I-like activity is associated with a soluble Yck2-beta-galact...

  4. Effect of Fluoride, Casein Phosphopeptide–Amorphous Calcium Phosphate and Casein Phosphopeptide–Amorphous Calcium Phosphate Fluoride on Enamel Surface Microhardness After Microabrasion: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Ghazaleh Ahmadi Zenouz

    2016-03-01

    Full Text Available Objectives: This study aimed to assess the effect of applying casein phosphopeptide–amorphous calcium phosphate (CPP-ACP paste, casein phosphopeptide–amorphous calcium phosphate fluoride (CPP-ACPF paste and sodium fluoride gel on surface microhardness of enamel after microabrasion.Materials and Methods: Thirty freshly extracted human premolars were selected. All samples were subjected to hardness indentations made with the Vickers hardness machine and the average value was recorded as the initial surface microhardness. The specimens were then randomly divided into three groups (n=10 of CPP-ACPF, fluoride and CPP-ACP. The teeth were micro-abraded with Opalustre. Microhardness test was performed to assess the post-abrasion hardness. Three remineralization modalities were performed on samples of each group. The enamel surface microhardness measurements were performed. To compare the difference between groups, the rehardening and softening values were defined. One-way ANOVA and Tukey’s post hoc test at a significance level of 5% were used for statistical analysis.Results: The mean microhardness value (MMV had a significant decrease after microabrasion from baseline. The MMV had a significant increase after remineralization in all groups. The MMV of CPP-ACPF group was significantly more than that of fluoride group (P=0.027. The rehardening value of fluoride group was significantly more than that of other groups (P<0.001.Conclusion: All the remineralizing agents were effective for rehardening the enamel after microabrasion. The CPP-ACP and CPP-ACPF pastes are effective, but to a lesser extent than neutral sodium fluoride gel in remineralizing enamel surface. Incorporation of fluoride to CPP-ACP formulation does not provide any additional remineralizing potential.Keywords: Casein phosphopeptide-amorphous calcium phosphate nanocomplex; Enamel Microabrasion; Hardness; Sodium Fluoride

  5. Casein addition to a whey-based formula has limited effects on gut function in preterm pigs

    DEFF Research Database (Denmark)

    Thymann, T.; Støy, Ann Cathrine Findal; Bering, S. B.;

    2012-01-01

    studied in gut contents. Severity of NEC lesions was similar between diet groups but galactose absorption was markedly higher in CASEIN than in WHEY (P <0.01) although only 6 h after the start of the enteral feeding period. There were no differences in ex vivo 14C-D-glucose uptake, digestive enzymes...... followed by 30 h of enteral feeding with whey [protein fraction of milk formula based on whey (WHEY); n = 11] or casein and/or whey [protein fraction of milk formula based on a combination of casein and whey (CASEIN); n = 9]-based formulas. Sugar absorptive function was investigated at 6 and 30 h after...... initiation of enteral feeding using bolus feedings with galactose and mannitol. Pigs were killed after the last in vivo sugar absorption test and evaluated for NEC and the mid intestine was used for ex vivo measurements of hexose absorption. Microbiota profile and short chain fatty acid (SCFA) levels were...

  6. Caseinolytic activity of fruit extract from Opuntia ficus-indica on bovine, caprine, and ovine Sodium Caseinates

    OpenAIRE

    Pintado, Ana I.; Macedo, Angela C.; Teixeira, Grimaneza; Pais, Salomé; Clemente, Alda; Malcata, F. Xavier

    2001-01-01

    The rates and extents of hydrolysis of RS- and â-caseins from bovine, caprine, and ovine sodium caseinates produced by an enzymatic extract of the fruit of Opuntia ficusindica, (L.) Miller were evaluated and compared with those produced by a commercial animal rennet. A mechanistic model based on a pseudo-first-order enzymatic reaction, in the presence of first-order deactivation of the enzyme, was postulated and successfully fitted to the experimental data. The animal rennet ex...

  7. Hydrolysis of Casein Micelles by AprX Enzym from Pseudomonas fluorescens Induces their Déstabilisation

    OpenAIRE

    Baglinière, François; Mateos, A. (Ana); Jardin, Julien; Rousseau, Florence; Robert, Benoit; Humbert, G.; Dary, A.; Gaillard, J L; Amiel, C

    2012-01-01

    Casein micelle is a colloidal structure containing different casein molecules and calcium phosphate. Different forces are involved in this micellar organisation and contribute to their stability. However micellar destabilisations can be observed after modifications of physico-chemical conditions like acidification, calcium addition, intensive heat treatments, high pressure or proteolysis with as consequence the formation of a gel or a sediment. The objective of this work was to understand th...

  8. Influence of calcium chelators on concentrated micellar casein solutions : from micellar structure to viscosity and heat stability

    OpenAIRE

    Kort, de, YAW Yvonne

    2012-01-01

    In practice it is challenging to prepare a concentrated medical product with high heat stability and low viscosity. Calcium chelators are often added to dairy products to improve heat stability, but this may increase viscosity through interactions with the casein proteins. The aim of this thesis was to obtain a better understanding of the influence of different calcium chelators on the physico-chemical properties of casein micelles and the resulting effect on viscosity and heat stability of c...

  9. Transfection of beta-casein chimeric gene and hormonal induction of its expression in primary murine mammary epithelial cells.

    OpenAIRE

    Yoshimura, M.; Oka, T

    1990-01-01

    To study the regulatory sequence elements responsible for casein gene expression, we constructed a chimeric gene containing 5.3 kilobases (kb) of the 5'-flanking sequence and 1.6 kb of the 3'-flanking sequence of the mouse beta-casein gene fused to the bacterial chloramphenicol acetyl-transferase (CAT) gene. The chimeric gene was transfected by the calcium phosphate-precipitation procedure into primary mouse mammary epithelial cells prepared from pregnant mice. The transfection procedure had ...

  10. Allelic Frequency of Kappa-Casein and Beta-Lactoglobulin in Indian Crossbred (Bos taurus X Bos indicus) Dairy Bulls

    OpenAIRE

    Patel, Rajesh K.; Chauhan, Jenabhai B.; Singh, Krishna M.; Kalpesh J. Soni

    2007-01-01

    A polymerase chain reaction - restriction fragment length polymorphism test was performed on DNA samples extracted from blood samples of 256 Holstein Friesian crossbred and 112 from Jersey crossbred bulls/bull calves to detect allelic frequency of the bovine kappa-casein and beta-lactoglobulin gene responsible for milk production traits. A 350 bp fragment of kappa-casein and a 247 bp fragment of beta-lactoglobulin were amplified and digested with Hinf I and Hae III restriction enzymes, respec...

  11. Coagulation properties and Nostrano di Primiero cheese yield of milk from Brown grazing cows of different k-casein genotype

    OpenAIRE

    G. Gaiarin; W. Ventura; S. Bovolenta; E. Saccà

    2011-01-01

    Genetic variants of milk proteins affect composition, technological characteristics and, as a consequence, cheesemaking properties of milk (Jakob and Puhan, 1992; Mariani and Summer, 1999). The effects of αS1, β and k-caseins variants, on milk coagulation properties are well known (Grosclaude, 1988). Cheesemaking involving many cheese varieties showed shorter clotting and curd firming times, higher curd firmness and cheese yields with the k-casein BB variant (Van Den Berg et al., 1...

  12. Mesenteric IL-10-producing CD5(+) regulatory B cells suppress cow's milk casein-induced allergic responses in mice.

    Science.gov (United States)

    Kim, A-Ram; Kim, Hyuk Soon; Kim, Do Kyun; Nam, Seung Taek; Kim, Hyun Woo; Park, Young Hwan; Lee, Dajeong; Lee, Min Bum; Lee, Jun Ho; Kim, Bokyung; Beaven, Michael A; Kim, Hyung Sik; Kim, Young Mi; Choi, Wahn Soo

    2016-01-01

    Food allergy is a hypersensitive immune reaction to food proteins. We have previously demonstrated the presence of IL-10-producing CD5(+) B cells and suggested their potential role in regulating cow's milk casein allergy in humans and IgE-mediated anaphylaxis in mice. In this study, we determined whether IL-10-producing CD5(+) regulatory B cells control casein-induced food allergic responses in mice and, if so, the underlying mechanisms. The induction of oral tolerance (OT) by casein suppressed casein-induced allergic responses including the decrease of body temperature, symptom score, diarrhea, recruitment of mast cells and eosinophils into jejunum, and other biological parameters in mice. Notably, the population of IL-10-producing CD5(+) B cells was increased in mesenteric lymph node (MLN), but not in spleen or peritoneal cavity (PeC) in OT mice. The adoptive transfer of CD5(+) B cells from MLN, but not those from spleen and PeC, suppressed the casein-induced allergic responses in an allergen-specific and IL-10-dependent manner. The inhibitory effect of IL-10-producing CD5(+) B cells on casein-induced allergic response was dependent on Foxp3(+) regulatory T cells. Taken together, mesenteric IL-10-producing regulatory B cells control food allergy via Foxp3(+) regulatory T cells and could potentially act as a therapeutic regulator for food allergy. PMID:26785945

  13. Moleculer analysis of genotype kappa casein and composition of goat milk Etawah grade, Saanen and their crossbred

    Directory of Open Access Journals (Sweden)

    Yayu Zurriyati

    2011-03-01

    Full Text Available Polymorphism of goat casein gene closely linked to the quality of milk protein. κ-casein is one of the casein fractions that influence the shape and stability of grain milk. This study is aimed to identify the variation of genotype κ-casein and related with milk quality from Etawah grade, Saanen and their crossbreed (PE-SA. The number of dairy goats used in this study was 150 animals consisted of Etawah grade (48 animals, Saanen (51 animals and PE-SA (51 animals. Steps of experiment were: blood and milk sampling collection, DNA amplification by PCR and the product digestion using Pst1 enzyme, κ-casein gene sequencing and analyzing the quality of fresh milk. The results showed that κ-casein gene is monomorphic by PCR-RFLP (Pst1 for all the goat breeds, but DNA sequencing indicated 38 point of mutation. Observation on goat milk quality showed that Etawah grade milk had highest (P < 0.05 density value (1.033 ± 0.002 and solid non fat (9.577 ± 0.704% than those of Saanen and PE-SA fresh milk goat.

  14. Casein improves brachial and central aortic diastolic blood pressure in overweight adolescents: a randomised, controlled trial

    DEFF Research Database (Denmark)

    Arnberg, Karina; Larnkjær, Anni; Michaelsen, Kim F.;

    2013-01-01

    Arterial stiffness, blood pressure (BP) and blood lipids may be improved by milk in adults and the effects may be mediated via proteins. However, limited is known about the effects of milk proteins on central aortic BP and no studies have examined the effects in children. Therefore, the present...... stiffness or blood lipid concentrations. A high intake of casein improves DBP in overweight adolescents. Thus, casein may be beneficial for younger overweight subjects in terms of reducing the longterm risk of CVD. In contrast, whey protein seems to increase BP compared with drinking water; however, water...... trial examined the effect of milk and milk proteins on brachial and central aortic BP, blood lipids, inflammation and arterial stiffness in overweight adolescents. A randomised controlled trial was conducted in 193 overweight adolescents aged 12–15 years. They were randomly assigned to drink 1 litre of...

  15. Disorder in Milk Proteins: Formation, Structure, Function, Isolation and Applications of Casein Phosphopeptides.

    Science.gov (United States)

    Naqvi, Muhammad Ali; Irani, Kimia Anaraki; Katanishooshtari, Maryam; Rousseau, Dérick

    2016-01-01

    This article is a continuation of a series of reviews on the presence and the role of intrinsic disorder in milk proteins in the journal of Current Protein and Peptide Science. The focus of this article is on casein phosphopeptides, which are liberated during digestion of the milk protein casein. Structurally these phosphopeptides have multiphosphorylated regions making them highly charged. The high degree of charge coupled with relatively low instances of hydrophobic amino acids makes them intrinsically disordered. These peptides have anticariogenic, antimicrobial, immunomodulatory, and cytomodulatory properties. Recent work using in vivo and in vitro models suggests that in addition to transporting calcium, these peptides can also enhance its bioaccessibility. The mechanism of this enhancement has yet to be determined. We review the current state of their structure, function, and isolation of these peptides. PMID:26630980

  16. ANALYSIS OF POLYMORPHISM OF ALPHA S1 CASEIN OF SLOVAK PINZGAU CATTLE BY PCR-RFLP

    Directory of Open Access Journals (Sweden)

    MARTINA MILUCHOVÁ

    2013-07-01

    Full Text Available The work was oriented to identification of -s1 casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 93 cattle. Bovine genomic DNA was isolated by fenol-chlorophorm deprotenization and ethanol precipitation and used in order to estimate -s1 casein genotypes by means of PCR-RFLP method. The PCR products were digested with MaeIII restriction enzyme. In the population included in the study there were homozygote genotype BB (81 animals and heterozygote genotype BC (12 animals. Homozygote genotype CC has not been observed. In the total population of cattle homozygotes BB – 0.871 were the most frequent, while BC – 0.129 were the least frequent ones. This suggests a superiority of allele B – 0.9355.

  17. Genetic Structure of Alpha S1 Casein in Slovak Pinzgau Cattle

    Directory of Open Access Journals (Sweden)

    Anna Trakovická

    2011-05-01

    Full Text Available The work was oriented to identification of -s1 casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 39 cattle. Bovine genomic DNA was isolated by commercial kit NucleoSpin Blood (Macherey-Nagel and ethanol precipitation and used in order to estimate -s1 casein genotypes by means of PCR-RFLP method. The PCR products were digested with MaeIII restriction enzyme. In the population included in the study, homozygote genotype BB (39 animals and heterozygote genotype BC (3 animals. Homozygote genotype CC has not been observed. In the total population of cattle homozygotes BB – 0.9231 were the most frequent, while BC – 0.0769 were the least frequent ones. This suggests a superiority of allele B – 0.9615.

  18. Water ingress into a casein film quantified using time-resolved neutron imaging.

    Science.gov (United States)

    Metwalli, E; Hermes, H E; Calzada, E; Kulozik, U; Egelhaaf, S U; Müller-Buschbaum, P

    2016-03-01

    The migration of water into a casein film was probed with neutron radiography. From the neutron transmission images, the evolution of the water saturation profiles was extracted. The results indicate that the water influx is dominated by imbibition but also contains a diffusional component. The time dependence of the water ingress was quantified using a diffusion-like equation previously also applied to imbibition. A water transport coefficient D = 0.9 × 10(-9) m(2) s(-1) was found. This value and direct observation of the images indicate that the time taken for a typical adhesive casein-based layer to become saturated with water is of the order of hours. PMID:26862596

  19. α-Casein Inhibits Insulin Amyloid Formation by Preventing the Onset of Secondary Nucleation Processes.

    Science.gov (United States)

    Librizzi, Fabio; Carrotta, Rita; Spigolon, Dario; Bulone, Donatella; San Biagio, Pier Luigi

    2014-09-01

    α-Casein is known to inhibit the aggregation of several proteins, including the amyloid β-peptide, by mechanisms that are not yet completely clear. We studied its effects on insulin, a system extensively used to investigate the properties of amyloids, many of which are common to all proteins and peptides. In particular, as for other proteins, insulin aggregation is affected by secondary nucleation pathways. We found that α-casein strongly delays insulin amyloid formation, even at extremely low doses, when the aggregation process is characterized by secondary nucleation. At difference, it has a vanishing inhibitory effect on the initial oligomer formation, which is observed at high concentration and does not involve any secondary nucleation pathway. These results indicate that an efficient inhibition of amyloid formation can be achieved by chaperone-like systems, by sequestering the early aggregates, before they can trigger the exponential proliferation brought about by secondary nucleation mechanisms. PMID:26278257

  20. Phosphorylation in vitro of human fibrinogen with casein kinase TS and characterization of phosphorylated sites

    International Nuclear Information System (INIS)

    Human fibrinogen was phosphorylated by casein kinase TS. The [32P]phosphate incorporated varied between 0.5 and 1 mol of phosphate per mole of fibrinogen. The phosphate was localized to Ser523 and Ser590 and serine and threonine residues between amino acids 259 and 268 in the A alpha-chain. In addition, Thr416 and Ser420 were phosphorylated in the gamma'-chain, which is a variant of the gamma-chain, constituting 7-10% of the gamma-chain population. The functional significance of casein kinase TS-induced phosphorylation of fibrinogen remains unknown; however, a slight but consistent increase of the turbidity in a gelation assay was observed for phosphorylated compared to unphosphorylated fibrinogen

  1. Enhancing proliferation and osteogenic differentiation of HMSCs on casein/chitosan multilayer films.

    Science.gov (United States)

    Li, Yan; Zheng, Zebin; Cao, Zhinan; Zhuang, Liangting; Xu, Yong; Liu, Xiaozhen; Xu, Yue; Gong, Yihong

    2016-05-01

    Creating a bioactive surface is important in tissue engineering. Inspired by the natural calcium binding property of casein (CA), multilayer films ((CA/CS)n) with chitosan (CS) as polycation were fabricated to enhance biomineralization, cell adhesion and differentiation. LBL self-assembly technique was used and the assembly process was intensively studied based on changes of UV absorbance, zeta potential and water contact angle. The increasing content of chitosan and casein with bilayers was further confirmed with XPS and TOF-SIMS analysis. To improve the biocompatibility, gelatin was surface grafted. In vitro mineralization test demonstrated that multilayer films had more hydroxyapatite crystal deposition. Human mesenchymal stem cells (HMSCs) were seeded onto these films. According to fluorescein diacetate (FDA) and cell cytoskeleton staining, MTT assay, expression of osteogenic marker genes, ALP activity, and calcium deposition quantification, it was found that these multilayer films significantly promoted HMSCs attachment, proliferation and osteogenic differentiation than TCPS control. PMID:26895501

  2. Remineralization of early enamel lesions using casein phosphopeptide amorphous calcium Phosphate: An ex-vivo study

    OpenAIRE

    Ruchi Vashisht; Anil Kumar; Indira, R; Srinivasan, M R; Ramachandran, S.

    2010-01-01

    Objective: This study aimed at qualitatively evaluating the remineralization potential of casein phosphopeptide amorphous calcium phosphate on artificial early enamel lesions in an ex-vivo scenario by observing the treated tooth surface using a scanning electron microscope (SEM). Materials and Methods: This randomized study was conducted on 10 subjects undergoing orthodontic treatment with premolar extraction as part of their treatment. Artificial white lesions were created with the applicati...

  3. Protective potential of casein phosphopeptide amorphous calcium phosphate containing paste on enamel surfaces

    OpenAIRE

    Padmini Somasundaram; Vimala, N; Lalita Gauri Mandke

    2013-01-01

    Background: Dental caries remains the most common dental disease facing mankind. Prevention of initiation and interruption in progression of early lesions are the desirable modes of caries management. There is a scope for agents, which may be used to enhance anti - caries activity. This need has redirected research to develop novel preventive agents that can act as an adjunct to fluoride or independent of it. Casein Phosphopeptide - Amorphous Calcium Phosphate (CPP-ACP) is one such agent that...

  4. Activation of mammalian DNA ligase I through phosphorylation by casein kinase II.

    OpenAIRE

    Prigent, C.; Lasko, D D; Kodama, K.; Woodgett, J R; Lindahl, T

    1992-01-01

    Mammalian DNA ligase I has been shown to be a phosphoprotein. Dephosphorylation of purified DNA ligase I causes inactivation, an effect dependent on the presence of the N-terminal region of the protein. Expression of full-length human DNA ligase I in Escherichia coli yielded soluble but catalytically inactive enzyme whereas an N-terminally truncated form expressed activity. Incubation of the full-length preparation from E. coli with purified casein kinase II (CKII) resulted in phosphorylation...

  5. Hematein, a casein kinase II inhibitor, inhibits lung cancer tumor growth in a murine xenograft model

    OpenAIRE

    Hung, Ming-Szu; Xu, Zhidong; Chen, Yu; Smith, Emmanuel; Mao, Jian-Hua; Hsieh, David; Lin, Yu-Ching; Yang, Cheng-Ta; Jablons, David M.; You, Liang

    2013-01-01

    Casein kinase II (CK2) inhibitors suppress cancer cell growth. In this study, we examined the inhibitory effects of a novel CK2 inhibitor, hematein, on tumor growth in a murine xenograft model. We found that in lung cancer cells, hematein inhibited cancer cell growth, Akt/PKB Ser129 phosphorylation, the Wnt/TCF pathway and increased apoptosis. In a murine xenograft model of lung cancer, hematein inhibited tumor growth without significant toxicity to the mice tested. Molecular docking showed t...

  6. The adsorption of lysozyme and chymosin onto emulsion droplets and their association with caseins

    OpenAIRE

    Roos, de, S.A.

    1999-01-01

    The proteolytic action of proteases present in cheese plays a major role in the ripening of cheese. These proteases originate from the rennet, the starter cultures and from the milk itself. The proteolysis in cheese results in the degradation of the casein proteins into smaller peptides and free amino acids, which act as flavour precursors. The ripening of cheese under conditioned storage is time consuming and costly. Addition of specific enzymes to the cheese milk is one of several options t...

  7. Casein kinase I phosphorylates and destabilizes the β-catenin degradation complex

    OpenAIRE

    Gao, Zhong-Hua; Seeling, Joni M.; Hill, Virginia; Yochum, April; Virshup, David M.

    2002-01-01

    Wnt signaling plays a key role in cell proliferation and development. Recently, casein kinase I (CKI) and protein phosphatase 2A (PP2A) have emerged as positive and negative regulators of the Wnt pathway, respectively. However, it is not clear how these two enzymes with opposing functions regulate Wnt signaling. Here we show that both CKIδ and CKIɛ interacted directly with Dvl-1, and that CKI phosphorylated multiple components of the Wnt-regulated β-catenin degradation complex in vitro, inclu...

  8. A Milk Protein, Casein, as a Proliferation Promoting Factor in Prostate Cancer Cells

    OpenAIRE

    Park, Sung-Woo; Kim, Joo-Young; Kim, You-sun; Lee, Sang Jin; Lee, Sang Don; CHUNG, MOON KEE

    2014-01-01

    Purpose Despite most epidemiologic studies reporting that an increase in milk intake affects the growth of prostate cancer, the results of experimental studies are not consistent. In this study, we investigated the proliferation of prostate cancer cells treated with casein, the main protein in milk. Materials and Methods Prostate cancer cells (LNCaP and PC3), lung cancer cells (A459), stomach cancer cells (SNU484), breast cancer cells (MCF7), immortalized human embryonic kidney cells (HEK293)...

  9. How to Squeeze a Sponge: Casein Micelles under Osmotic Stress, a SAXS Study

    OpenAIRE

    Bouchoux, Antoine; Gésan-Guiziou, Geneviève; Pérez, Javier; Cabane, Bernard

    2010-01-01

    By combining the osmotic stress technique with small-angle x-ray scattering measurements, we followed the structural response of the casein micelle to an overall increase in concentration. When the aqueous phase that separates the micelles is extracted, they behave as polydisperse repelling spheres and their internal structure is not affected. When they are compressed, the micelles lose water and shrink to a smaller volume. Our results indicate that this compression is nonaffine, i.e., some p...

  10. Genetic polymorphism of the K-casein (CSN3) gene in goats reared in Southern Italy

    OpenAIRE

    Fabio Pilla; Antonella Angiolillo; Armand Sanchez; Josè Maria Folch; Mohamed Habib Yahyaoui; Silvia Reale

    2010-01-01

    K-casein (K-CN) represents one of the most important proteins determining the manufacturing properties of milk,because of its essential role in micelle formation and stabilisation.Several genetic variants of K-CN have been described in goats. To investigate the occurrence of seven alleles and theirdistribution among breeds, a total of 170 animals, from six different breeds reared in Italy (Cilentana Nera, Derivata diSiria, Maltese, Jonica, Garganica and Cashmere), have been analysed in this p...

  11. Does ultrafiltration have a lasting effect on the physico-chemical properties of the casein micelles?

    OpenAIRE

    Ferrer, Mary Ann; Alexander, Marcela; Corredig, Milena

    2011-01-01

    The objective of this work was to determine if concentration of milk using ultrafiltration (in the absence of diafiltration) affects the physico-chemical properties of the casein micelles. The milk, once concentrated, was brought back to its original concentration and the physico-chemical properties as well as its susceptibility to rennet induced gelation were assessed. Although much is understood on the renneting behavior of concentrated milk, no information is available on how (or if) ultra...

  12. Casein nanoparticles as carriers for the oral delivery of folic acid

    OpenAIRE

    Peñalva, R. (Rebeca); Esparza, I; Agüeros, M.; Gonzalez-Navarro, C. (Carlos); Gonzalez-Ferrero, C. (Carolina); J.M. Irache

    2014-01-01

    Alimentary proteins can be viewed as an adequate material for the preparation of nanoparticles and microparticles. They offer several advantages such as their digestibility, price and a good capability to interact with a wide variety of compounds and nutrients. The aim of this work was to prepare and characterize casein nanoparticles for the oral delivery of folic acid. These nanoparticles were prepared by a coacervation process, stabilized with either lysine or arginine and, f...

  13. Remineralization of early enamel lesions using casein phosphopeptide amorphous calcium Phosphate: An ex-vivo study

    Directory of Open Access Journals (Sweden)

    Ruchi Vashisht

    2010-01-01

    Full Text Available Objective: This study aimed at qualitatively evaluating the remineralization potential of casein phosphopeptide amorphous calcium phosphate on artificial early enamel lesions in an ex-vivo scenario by observing the treated tooth surface using a scanning electron microscope (SEM. Materials and Methods: This randomized study was conducted on 10 subjects undergoing orthodontic treatment with premolar extraction as part of their treatment. Artificial white lesions were created with the application of 37% phosphoric acid for 20 mins. Teeth were then divided into two groups: one experimental and the other control. Customised orthodontic band with a window was luted with intermediate restorative material in the experimental group whereas in the control group, band without a window was luted. The casein phosphopeptide amorphous calcium phosphate (GC TOOTH MOUSSE paste was then applied on the window region of the experimental group for 3 mins thrice daily after meals for 14 days, whereas no paste was applied in the control group. After 14 days, teeth were extracted and viewed under an SEM. Results: The study groups showed remineralization of the lesions as compared with the control group in most of the samples. Conclusion: Casein phophopeptide could significantly remineralize the artificial enamel lesions in vivo.

  14. Effects of calcium chelators on calcium distribution and protein solubility in rennet casein dispersions.

    Science.gov (United States)

    McIntyre, Irene; O' Sullivan, Michael; O' Riordan, Dolores

    2016-04-15

    This study investigated the effects of calcium chelating salts on calcium-ion activity (ACa(++)), calcium distribution, and protein solubility in model CaCl2 solutions (50 mmol L(-1)) or rennet casein dispersions (15 g/100 g). Disodium phosphate and trisodium citrate at concentrations of 10 and 30 mmol L(-1) and at ratios of 1:0, 2:1, 1:1, 1:2 and 0:1 were added to both systems. The CaCl2 system, despite its simplicity, was a good indicator of chelating salt-calcium interactions in rennet casein dispersions. Adding trisodium citrate either alone or as part of a mixed chelating salt system resulted in high levels of dispersed "chelated" calcium; conversely, disodium phosphate addition resulted in lower levels, while the ACa(++) decreased with increasing concentration of both chelating salts. Neither chelating salt produced high levels of soluble protein. Thus calcium chelating salts may play a more subtle role in modulating hydration during manufacture of casein-based matrices than simply solubilising calcium or protein. PMID:26616945

  15. Gluten- and casein-free dietary intervention for autism spectrum conditions.

    Directory of Open Access Journals (Sweden)

    Paul eWhiteley

    2013-01-01

    Full Text Available Dietary intervention as a tool for maintaining and improving physical health and wellbeing is a widely researched and discussed topic. Speculation that diet may similarly affect mental health and wellbeing particularly in cases of psychiatric and behavioural symptomatology opens up various avenues for potentially improving quality of life. We examine evidence suggestive that a gluten-free, casein-free or combined gluten- and casein-free diet can ameliorate core and peripheral symptoms and improve developmental outcome in some cases of autism spectrum conditions. Although not wholly affirmative, the majority of published studies indicate statistically significant positive changes to symptom presentation following dietary intervention. In particular, changes to areas of communication, attention and hyperactivity are detailed, despite the presence of various methodological shortcomings. Specific characteristics of best- and non-responders to intervention have not been fully elucidated; neither has the precise mode of action for any universal effect outside of known individual cases of food-related co-morbidity. With the publication of controlled medium- and long-term group studies of a gluten- and casein-free diet alongside more consolidated biological findings potentially linked to intervention, the appearance of a possible diet-related autism phenotype seems to be emerging supportive of a positive dietary effect in some cases. Further debate on whether such dietary intervention should form part of best practice guidelines for autism spectrum conditions and onward representative of an autism dietary-sensitive enteropathy is warranted.

  16. Antioxidative effect of yak milk caseinates hydrolyzed with three different proteases

    Directory of Open Access Journals (Sweden)

    Santosh Kumar

    2013-09-01

    Full Text Available Aim: Yak milk is a type of milk that people are less familiar with due to its remote geographical location which may havesignificant effects on composition, microbiota and hydrolytic outcome. Present work was designed with the aim to evaluatethe antioxidative effect of peptides derived from yak milk caseinate on hydrolysis with three different proteases.Materials and Methods: In this investigation Yak milk casein was hydrolyzed by three commercially available proteases(Trypsin, Pepsin and chymotrypsin. These hydrolysates collected at different hydrolysis times (30 min, 60 min, 90 min, 120min, 150 min, 180 min, 210 min, 240 min, 270 min, 300 min, 330 min and 360 min were assayed for their antioxidant activitywith respect to the effect of incubation period.Results: Among all the enzyme hydrolysates, the tryptic hydrolysates showed highest antioxidant activity followed bychymotryptic hydrolysates. Further, the peptide samples showing highest activity were subjected to RP-HPLC for their partialcharacterization. Tryptic and peptic hydrolysates produced peaks mainly in the region of hydrophillic solvent indicating thepresence of hydrophillic peptides/peptides.Conclusion: The results indicated that yak milk casein could be a resource to generate antioxidative peptides and be used asmultifunctional active ingredients for many value-added functional foods as well as a traditional food protein.

  17. Genetic parameters for casein and urea content in the Italian Brown Swiss dairy cattle

    Directory of Open Access Journals (Sweden)

    O. Pedron

    2010-04-01

    Full Text Available A total of 137,753 test day records of 20,745 Italian Brown Swiss dairy cows from 26 provinces of Italy were used to estimate heritability for casein and urea content in milk and their genetic correlations with other production traits and milk somatic cell score. Milk component values were obtained by Fourier Transformed Infrared (IR Spectroscopy from milk samples collected during national routine recording and were analysed using test day repeatability animal models. Fixed effects included 1,001 levels of herd-test date, 15 classes of days in milk, and 13 classes of age at calving within parity. The variation among cows was large for most of the traits. The heritability value for casein content was 0.31, as for protein content, and genetic and phenotypic correlations between these two traits were large (0.99 and 0.97 respectively. Milk urea content had a heritability of 0.17 and a positive genetic relationship with fat (0.12, null with protein (0.03 and casein (0.002 content and a negative genetic correlation with milk yield (-0.17 suggesting that the genetic improvement for milk urea content would be possible, but genetic gain would be affected by other traits included as selection criteria in the economic index and by their relative economic emphasis.

  18. Extracellular thermostable proteolytic activity of the milk spoilage bacterium Pseudomonas fluorescens PS19 on bovine caseins.

    Science.gov (United States)

    Stuknytė, M; Decimo, M; Colzani, M; Silvetti, T; Brasca, M; Cattaneo, S; Aldini, G; De Noni, I

    2016-06-01

    We studied the thermostable proteolytic activity of Pseudomonas fluorescens PS19 isolated from raw bovine milk. The heat-treated cell-free supernatant (HT-CFS) contained a thermostable protease of approximately 45 kDa, as revealed by casein zymography. We assigned this enzyme to P. fluorescens AprX metalloprotease (UniProtKB Acc. No. C9WKP6). After concentration by ultrafiltration at 10 kDa, the HT-CFS showed 2 other thermostable proteolytic bands on zymogram, with molecular masses of approximately 15 and 25 kDa. The former resulted a fragment of the AprX protease, whereas the 25-kDa protease was not homologous to any known protein of Pseudomonas spp. Subsequently, we assessed the proteolytic activity of the HT-CFS on bovine αS-, β-, and κ-casein during in vitro incubation at 7 or 22°C. By means of ultra-performance liquid chromatography-tandem mass spectrometry we identified the released peptides (n=591). Some of them resisted proteolysis during the whole incubation period at both incubation temperatures and, therefore, they could be assumed as indicators of the proteolytic action of P. fluorescens PS19 on bovine caseins. PMID:26995139

  19. Incorporation of radiolabeled whey proteins into casein micelles by heat processing

    International Nuclear Information System (INIS)

    Skim milk was heated at .70, 95, and 140 degree C to simulate the processes of pasteurization, forewarming, and UHT sterilization, and the specific interactions between α-lactalbumin or β-lactoglobulin and the caseins studied using tracer amounts of added 14C-labeled whey protein. Radioactivities of the whey and of the washed casein pellets from renneted skim milk were measured and the extent of the interaction estimated. Upon heating skim milk at 70 degree C for 45 s, less than 2% β-lactoglobulin and less than .3% α-lactalbumin were incorporated into the curd. Heating at 95 degree C for .5 to 20 min resulted in 58 to 85% of the β-lactoglobulin and 8 to 55% of the α-lactalbumin becoming associated with the curd. Heating at 140 degree C for 2 and 4 s caused 43 and 54% of the β-lactoglobulin and 9 and 12% of the α-lactalbumin, respectively, to be bound to the curd fraction. The radiolabeling technique is very sensitive and useful for tracing low levels of interaction between whey proteins and casein in heated milk systems

  20. Structure and rheological behavior of casein micelle suspensions during ultrafiltration process

    Science.gov (United States)

    Pignon, F.; Belina, G.; Narayanan, T.; Paubel, X.; Magnin, A.; Gésan-Guiziou, G.

    2004-10-01

    The stability and mechanism underlying the formation of deposits of casein micelles during ultrafiltration process were investigated by small-angle and ultra small-angle x-ray scattering (SAXS and USAXS). The casein micelle dispersions consisted of phospho-caseinate model powders and the measurements probed length scales ranging from 1 to 2000 nm. Rheometric and frontal filtration measurements were combined with SAXS to establish the relationship between the rheological behavior of deposits (shear and/or compression) and the corresponding microstructure. The results revealed two characteristic length scales for the equilibrium structure with radius of gyrations Rg, about 100 and 5.6 nm pertaining to the globular micelles and their non-globular internal structure, respectively. The SAXS measurements further indicated that the increase of temperature from 20 to 70 °C or the decrease of pH from 6.6 to 6 lead to agglomeration of the globular micelles. In situ scattering measurements showed that the decrease of permeation flows is directly related to the deformation and compression of the micelles in the immediate vicinity of the membrane.

  1. Simulation of the shape and size of casein micelles in a film state.

    Science.gov (United States)

    Gebhardt, Ronald; Kulozik, Ulrich

    2014-04-01

    Size fractionated casein micelles (CMs) form homogeneous films in which they are densely packed. The lateral size of CMs in films can be well resolved by surface-sensitive methods, but the estimation of their heights is still a challenge. We show that height information can be obtained from scattering patterns of GISAXS experiments on highly ordered casein films. We use an elastic scattering approach within the distorted wave Born approximation (DWBA) to simulate for the first time the two-dimensional intensity distribution of a GISAXS experiment of the CM near their critical angle. The model which fits the GISAXS data best considers an ellipsoidal form factor for the CM and an arrangement on a hexagonal lattice. Our results indicate that during film formation the spherical solution structure of CMs becomes compressed in the direction perpendicular to the film surface. In the film state, the micelles assume an oblate ellipsoidal shape with an aspect ratio of 1.9. Hence, their surface and contact area to the surrounding increases. As a result, the density of κ-casein on the micellar surface decreases, which could influence the functional properties of coatings and films. PMID:24569528

  2. Trichostatin A Inhibits β-Casein Expression in Mammary Epithelial Cells

    Science.gov (United States)

    Pujuguet, Philippe; Radisky, Derek; Levy, Dinah; Lacza, Charlemagne; Bissell, Mina J.

    2010-01-01

    Many aspects of cellular behavior are defined by the content of information provided by association of the extracellular matrix (ECM) and with cell membrane receptors. When cultured in the presence of laminin-containing ECM and prolactin (Prl), normal mammary epithelial cells express the milk protein β-casein. We have previously found that the minimal ECM- and Prl-responsive enhancer element BCE-1 was only active when stably integrated into chromatin, and that trichostatin A (TSA), a reagent that leads to alterations in chromatin structure, was able to activate the integrated enhancer element. We now show that endogenous β-casein gene, which is controlled by a genetic assembly that is highly similar to that of BCE-1 and which is also activated by incubation in ECM and Prl, is instead inhibited by TSA. We provide evidence that the differing response of β-casein and BCE-1 to TSA is neither due to an unusual effect of TSA on mammary epithelial cells, nor to secondary consequences from the expression of a separate gene, nor to a particular property of the BCE-1 construct. As a component of this investigation, we also showed that ECM mediated rapid histone deacetylation in mammary epithelial cells. These results are discussed in combination with previous work showing that TSA mediates the differentiation of many types of cancer cells but inhibits differentiation of some nonmalignant cell types. PMID:11746508

  3. Casein Kinase 1 Epsilon Expression Predicts Poorer Prognosis in Low T-Stage Oral Cancer Patients

    Directory of Open Access Journals (Sweden)

    Shu-Hui Lin

    2014-02-01

    Full Text Available Casein kinase 1 is a group of ubiquitous serine/threonine kinases that are involved in normal cellular functions and several pathological conditions, such as DNA repair, cell cycle progression, cytokinesis, differentiation, and apoptosis. Recent studies have indicated that casein kinase 1-epsilon (CK1ε and casein kinase 1-delta (CK1δ expression has a role in human cancers. We investigated the associations between CK1ε and CK1δ expression and the clinical parameters of oral cancer using immunohistochemical study methods on oral squamous cell carcinoma specimens. The results of our immunohistochemical analysis showed that the loss of CK1ε expression was greatly associated with a poor four-year survival rate in oral cancer patients (p = 0.002. A Kaplan-Meier analysis showed that patients who had a loss of CK1ε expression had a considerably poorer overall survival rate than patients who had positive CK1ε expressions (p = 0.022. A univariate analysis revealed that patients who had a loss of CK1ε expression had considerably poorer overall survival (OS than patients who had positive expression (p = 0.024, hazard ratio (HR = 1.7. In conclusion, our data indicated that the loss of cytoplasmic CK1ε expression is greatly associated with poor survival and might be an adverse survival factor.

  4. The Role of B-Lactoglobulin in the Development of the Core-and-Lining Structure of Casein Particles in Acid-Heat-Induced Milk Gels

    OpenAIRE

    Harwalkar, V. R.; Kalab, Miloslav

    1988-01-01

    Acid-heat-induced gels were obtained by coagulating casein micelle dispersions at 90 C using glucono-and-lactone. The casein micelles used were isolated from raw skim milk by centrifugation, washed free of whey proteins and soluble salts, and dispersed in water or a milk dialyzate. The pH values of the gels varied from 4.7 to 6.3. A core-and-lining ultrastructure developed in casein particles coagulated at pH 5.2 to 5.5 from casein micelle dispersions in the milk dialyzate provided that B-lac...

  5. Interactions of milk α- and β-casein with malvidin-3-O-glucoside and their effects on the stability of grape skin anthocyanin extracts.

    Science.gov (United States)

    He, Zhiyong; Xu, Mingzhu; Zeng, Maomao; Qin, Fang; Chen, Jie

    2016-05-15

    The interactions of α- and β-casein with malvidin-3-O-glucoside (MG), the major anthocyanin in grape skin anthocyanin extracts (GSAE), were examined at pH 6.3 by fluorescence, fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopy. The binding constant (KS), binding force and effects of the interactions on the caseins conformation and GSAE stability were investigated. The results showed that α- and β-casein bound with MG via hydrophilic (van der Waals forces or hydrogen bonding) and hydrophobic interactions, respectively. α-Casein had a slightly stronger binding affinity toward MG than β-casein, with respective KS values of 0.51×10(3)M(-1) and 0.46×10(3)M(-1) at 297K. The secondary structures of α- and β-casein were changed by MG binding, with a decrease in α-helix and an increase in turn for α-casein and no change in α-helix and a decrease in turn for β-casein. The casein-anthocyanin interaction appeared to have a positive effect on the thermal, oxidation and photo stability of GSAE. PMID:26775977

  6. Studies on the photophysical properties of 1,8-naphthalimide derivative and aggregation induced emission recognition for casein

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Yang, E-mail: 66160692@qq.com [Department of Chemistry and Chemical Engineering, Xi' an University of Arts and Science, No. 168, Taibai South Road, Xi' an, Shaanxi 710065 (China); Liang, Xuhua; Fan, Jun [School of Chemical Engineering, Northwest University, No. 229, Taibai North Road, Xi' an, Shaanxi 710069 (China); Han, Quan, E-mail: xahanq@hotmail.com [Department of Chemistry and Chemical Engineering, Xi' an University of Arts and Science, No. 168, Taibai South Road, Xi' an, Shaanxi 710065 (China)

    2013-09-15

    A novel water-soluble 1,8-naphthalimide derivative 1, bearing two acetic carboxylic groups, exhibited fluorescent turn-on recognition for casein micelle based on the aggregation induced emission (AIE) character. The photophysical properties of 1 consisting of donor and acceptor units were investigated by the solvation effect. Changing from polar to non-polar solvent increased the solvent interaction; both the excitation and emission spectra were shifted to shorter wavelength and intensity decreased through taking advantage of twisted intramolecular charge transfer (TICT) and self-association fluorescence emission. Moreover, the red-shift and quenching in protic solvent were caused by the excited-state hydrogen bond strengthening effect. The density functional theory (DFT) and time dependent density functional theory (TDDFT) were used to obtain the most stable structure, electronic excitation energy, dipole moments and charge distribution. The AIE mechanism of 1 with casein micelle was due to 1 docked in the hydrophobic cavity between sub-micelles and bound with amino acid residues, resulting in the aggregation of 1 on the casein micelle surface and emission enhancement, based on which, a novel casein assay method was developed. The proposed method exhibited a good linear range from 0.1 to 10.5 μg mL{sup −1}, with the detection limit of 3.0 ng mL{sup −1}. Satisfactory reproducibility, reversibility and a short response time were realized. This method was applied for the determination of casein in milk powder samples, avoiding the interferences from other components and illegal additives in milk. -- Highlights: • A water-soluble 1,8-naphthalimide-based fluorescent probe 1 was synthesized. • Photophysical characterization of 1 was studied. • Aggregation induced emission enhancement of 1 with casein was investigated. • A novel casein quantification method was developed.

  7. Skeletal effect of casein and whey protein intake during catch-up growth in young male Sprague-Dawley rats.

    Science.gov (United States)

    Masarwi, Majdi; Gabet, Yankel; Dolkart, Oleg; Brosh, Tamar; Shamir, Raanan; Phillip, Moshe; Gat-Yablonski, Galia

    2016-07-01

    The aim of the present study was to determine whether the type of protein ingested influences the efficiency of catch-up (CU) growth and bone quality in fast-growing male rats. Young male Sprague-Dawley rats were either fed ad libitum (controls) or subjected to 36 d of 40 % food restriction followed by 24 or 40 d of re-feeding with either standard rat chow or iso-energetic, iso-protein diets containing milk proteins - casein or whey. In terms of body weight, CU growth was incomplete in all study groups. Despite their similar food consumption, casein-re-fed rats had a significantly higher body weight and longer humerus than whey-re-fed rats in the long term. The height of the epiphyseal growth plate (EGP) in both casein and whey groups was greater than that of rats re-fed normal chow. Microcomputed tomography yielded significant differences in bone microstructure between the casein and whey groups, with the casein-re-fed animals having greater cortical thickness in both the short and long term in addition to a higher trabecular bone fraction in the short term, although this difference disappeared in the long term. Mechanical testing confirmed the greater bone strength in rats re-fed casein. Bone quality during CU growth significantly depends on the type of protein ingested. The higher EGP in the casein- and whey-re-fed rats suggests a better growth potential with milk-based diets. These results suggest that whey may lead to slower bone growth with reduced weight gain and, as such, may serve to circumvent long-term complications of CU growth. PMID:27189324

  8. Studies on the photophysical properties of 1,8-naphthalimide derivative and aggregation induced emission recognition for casein

    International Nuclear Information System (INIS)

    A novel water-soluble 1,8-naphthalimide derivative 1, bearing two acetic carboxylic groups, exhibited fluorescent turn-on recognition for casein micelle based on the aggregation induced emission (AIE) character. The photophysical properties of 1 consisting of donor and acceptor units were investigated by the solvation effect. Changing from polar to non-polar solvent increased the solvent interaction; both the excitation and emission spectra were shifted to shorter wavelength and intensity decreased through taking advantage of twisted intramolecular charge transfer (TICT) and self-association fluorescence emission. Moreover, the red-shift and quenching in protic solvent were caused by the excited-state hydrogen bond strengthening effect. The density functional theory (DFT) and time dependent density functional theory (TDDFT) were used to obtain the most stable structure, electronic excitation energy, dipole moments and charge distribution. The AIE mechanism of 1 with casein micelle was due to 1 docked in the hydrophobic cavity between sub-micelles and bound with amino acid residues, resulting in the aggregation of 1 on the casein micelle surface and emission enhancement, based on which, a novel casein assay method was developed. The proposed method exhibited a good linear range from 0.1 to 10.5 μg mL−1, with the detection limit of 3.0 ng mL−1. Satisfactory reproducibility, reversibility and a short response time were realized. This method was applied for the determination of casein in milk powder samples, avoiding the interferences from other components and illegal additives in milk. -- Highlights: • A water-soluble 1,8-naphthalimide-based fluorescent probe 1 was synthesized. • Photophysical characterization of 1 was studied. • Aggregation induced emission enhancement of 1 with casein was investigated. • A novel casein quantification method was developed

  9. Identification of casein kinase 1, casein kinase 2, and cAMP-dependent protein kinase-like activities in Trypanosoma evansi

    Directory of Open Access Journals (Sweden)

    José Manuel Galán-Caridad

    2004-12-01

    Full Text Available Trypanosoma evansi contains protein kinases capable of phosphorylating endogenous substrates with apparent molecular masses in the range between 20 and 205 kDa. The major phosphopolypeptide band, pp55, was predominantly localized in the particulate fraction. Anti-alpha and anti-beta tubulin monoclonal antibodies recognized pp55 by Western blot analyses, suggesting that this band corresponds to phosphorylated tubulin. Inhibition experiments in the presence of emodin, heparin, and 2,3-bisphosphoglycerate indicated that the parasite tubulin kinase was a casein kinase 2 (CK2-like activity. GTP, which can be utilized instead of ATP by CK2, stimulated rather than inactivated the phosphorylation of tubulin in the parasite homogenate and particulate fraction. However, GTP inhibited the cytosolic CK2 responsible for phosphorylating soluble tubulin and other soluble substrates. Casein and two selective peptide substrates, P1 (RRKDLHDDEEDEAMSITA for casein kinase (CK1 and P2 (RRRADDSDDDDD for CK2, were recognized as substrates in T. evansi. While the enzymes present in the soluble fraction predominantly phosphorylated P1, P2 was preferentially labeled in the particulate fractions. These results demonstrated the existence of CK1-like and CK2-like activities primarily located in the parasite cytosolic and membranous fractions, respectively. Histone II-A and kemptide (LRRASVA also behaved as suitable substrates, implying the existence of other Ser/Thr kinases in T. evansi. Cyclic AMP only increased the phosphorylation of histone II-A and kemptide in the cytosol, demonstrating the existence of soluble cAMP-dependent protein kinase-like activities in T. evansi. However, no endogenous substrates for this enzyme were identified in this fraction. Further evidences were obtained by using PKI (6-22, a reported inhibitor of the catalytic subunit of mammalian cAMP-dependent protein kinases, which specifically hindered the cAMP-dependent phosphorylation of histone II

  10. Ser2 is the autophosphorylation site in the beta subunit from bicistronically expressed human casein kinase-2 and from native rat liver casein kinase-2 beta

    DEFF Research Database (Denmark)

    Boldyreff, B; James, P; Staudenmann, W; Issinger, O G

    1993-01-01

    Human casein kinase-2 (CK-2) subunits alpha and beta were bicistronically expressed in bacteria. The recombinant holoenzyme shared all investigated properties with the native CK-2 from mammalian sources (rat liver, Krebs II mouse ascites tumour cells). Contrary to recombinant human CK-2 produced by...... self-assembly in vitro, the bicistronically expressed beta subunit was autophosphorylated during formation of the holoenzyme in bacteria. Electrospray ionisation mass spectrometry (ESI) revealed Ser2 (second amino acid, first serine) as the only phosphate acceptor site. Kinetic data obtained with...... either the phosphorylated or the unphosphorylated form of CK-2 did not differ significantly, suggesting that the autophosphorylation had no influence on the kinetic parameters Km and Vmax. In parallel, native rat liver CK-2 beta subunit was shown to incorporate 0.1 mol phosphate/mol holoenzyme, which...

  11. Utilization in rats of 14C-L-lysine-labeled casein browned by amino-carbonyl reaction

    International Nuclear Information System (INIS)

    The investigation was carried out in order to elucidate the reason for the reduction in nutritive value of browned protein, by using labeled casein as a model protein. Goat casein preparation in which lysine residues had been labeled with 14C was browned by amino-carbonyl reaction with glucose at 370C. Browned or non-browned casein was ingested by growing rats by spaced feeding. When the rats ingested the browned casein the experimental group, higher radioactivity was found in TCA-soluble fraction in the small intestine as compared with that in the control group, while radioactivity was scarecely found in feces for 22 hr. Along with absorption delay, considerably high radioactivity was found in urine. The recovery of radioactivity in expired air of rats fed the labeled casein (browned and non-browned) was measured. In the experimental group, expired 14CO2 came out slower than the control group. From these results, it is suggested that the main reason for the reduction in nutritive value by browning reaction may be the formation of a lysine derivative in a protein, which remains in the small intestinal lumen as an absorption-delayed material and is finally excreted in urine. (auth.)

  12. Casein SNP in Norwegian goats: additive and dominance effects on milk composition and quality

    Directory of Open Access Journals (Sweden)

    Lien Sigbjørn

    2011-08-01

    Full Text Available Abstract Background The four casein proteins in goat milk are encoded by four closely linked casein loci (CSN1S1, CSN2, CSN1S2 and CSN3 within 250 kb on caprine chromosome 6. A deletion in exon 12 of CSN1S1, so far reported only in Norwegian goats, has been found at high frequency (0.73. Such a high frequency is difficult to explain because the national breeding goal selects against the variant's effect. Methods In this study, 575 goats were genotyped for 38 Single Nucleotide Polymorphisms (SNP located within the four casein genes. Milk production records of these goats were obtained from the Norwegian Dairy Goat Control. Test-day mixed models with additive and dominance fixed effects of single SNP were fitted in a model including polygenic effects. Results Significant additive effects of single SNP within CSN1S1 and CSN3 were found for fat % and protein %, milk yield and milk taste. The allele with the deletion showed additive and dominance effects on protein % and fat %, and overdominance effects on milk quantity (kg and lactose %. At its current frequency, the observed dominance (overdominance effects of the deletion allele reduced its substitution effect (and additive genetic variance available for selection in the population substantially. Conclusions The selection pressure of conventional breeding on the allele with the deletion is limited due to the observed dominance (overdominance effects. Inclusion of molecular information in the national breeding scheme will reduce the frequency of this deletion in the population.

  13. Comparative study of glycine, alanine or casein as inert nitrogen sources in endotoxemic rats.

    Science.gov (United States)

    Chambon-Savanovitch, C; Felgines, C; Farges, M C; Raul, F; Cézard, J P; Davot, P; Vasson, M P; Cynober, L A

    1999-10-01

    Pharmacological effects of dietary amino acids (AA) and peptides must be compared to an isonitrogenous control that is as inert as possible. To establish a rationale for the choice of such a control, potential metabolic and nutritional effects of three currently used nitrogenous controls (glycine, alanine, and casein) were evaluated in an endotoxemic rat model that has well-defined alterations in AA and protein metabolism. Five-week-old male Sprague-Dawley rats (113 +/- 1 g) were randomly assigned to four groups and received at d 0 an intraperitoneal injection of endotoxin (3 mg/kg). After withdrawal of food for 24 h, the rats were enterally refed for 48 h with a liquid diet (Osmolite((R))) supplemented with 0.19 g N. kg(-1). d(-1) in the form of glycine [lipopolysaccharide (LPS)-GLY group], alanine (LPS-ALA group) or casein (LPS-CAS group). One group (LPS group) received only Osmolite((R)). Plasma, two skeletal muscles, the liver and the intestine were then removed. Body and tissue weights and tissue protein contents did not differ among the four groups. Intestine histomorphometry showed no significant difference among groups. Jejunal hydrolase activities were significantly affected by the nitrogenous supplementations, but no effect was observed in the ileum. Only limited significant effects were observed on plasma and tissue-free AA concentrations, except for an accumulation of glycine in the plasma and tissues from the LPS-GLY group, compared to other groups. Overall, whereas glycine as a nitrogenous control should be used with care, either alanine or casein may be used as the "placebo," with the choice depending on the study to be performed. PMID:10498760

  14. Celecoxib Encapsulation in β-Casein Micelles: Structure, Interactions, and Conformation.

    Science.gov (United States)

    Turovsky, Tanya; Khalfin, Rafail; Kababya, Shifi; Schmidt, Asher; Barenholz, Yechezkel; Danino, Dganit

    2015-07-01

    β-Casein is a 24 kDa natural protein that has an open conformation and almost no folded or secondary structure, and thus is classified as an intrinsically unstructured protein. At neutral pH, β-casein has an amphiphilic character. Therefore, in contrast to most unstructured proteins that remain monomeric in solution, β-casein self-assembles into well-defined core-shell micelles. We recently developed these micelles as potential carriers for oral administration of poorly water-soluble pharmaceuticals, using celecoxib as a model drug. Herein we present deep and precise insight into the physicochemical characteristics of the protein-drug formulation, both in bulk solution and in dry form, emphasizing drug conformation, packing properties and aggregation state. In addition, the formulation is extensively studied in terms of structure and morphology, protein/drug interactions and physical stability. Particularly, NMR measurements indicated strong drug-protein interactions and noncrystalline drug conformation, which is expected to improve drug solubility and bioavailability. Small-angle X-ray scattering (SAXS) and cryogenic transmission electron microscopy (cryo-TEM) were combined for nanostructural characterization, proving that drug-protein interactions lead to well-defined spheroidal micelles that become puffier and denser upon drug loading. Dynamice light scattering (DLS), turbidity measurements, and visual observations complemented the analysis for determining formulation structure, interactions, and stability. Additionally, it was shown that the loaded micelles retain their properties through freeze-drying and rehydration, providing long-term physical and chemical stability. Altogether, the formulation seems greatly promising for oral drug delivery. PMID:26068530

  15. Characteristics of the interaction of calcium with casein submicelles as determined by analytical affinity chromatography

    International Nuclear Information System (INIS)

    Interaction of calcium with casein submicelles was investigated in CaCl2 and calcium phosphate buffers and with synthetic milk salt solutions using the technique of analytical affinity chromatography. Micelles that had been prepared by size exclusion chromatography with glycerolpropyl controlled-pore glass from fresh raw skim milk that had never been cooled, were dialyzed at room temperature against calcium-free imidazole buffer, pH 6.7. Resulting submicelles were covalently immobilized on succinamidopropyl controlled-pore glass (300-nm pore size). Using 45Ca to monitor the elution retardation, the affinity of free Ca2+ and calcium salt species was determined at temperatures of 20 to 40 degrees C and pH 6.0 to 7.5. Increasing the pH in this range or increasing the temperature strengthened the binding of calcium to submicelles, similar to previous observations with individual caseins. However, the enthalpy change obtained from the temperature dependence was considerably greater than that reported for alpha s1- and beta-caseins. Furthermore, the elution profiles for 45Ca in milk salt solutions were decidedly different from those in CaCl2 or calcium phosphate buffers and the affinities were also greater. For example, at pH 6.7 and 30 degrees C the average dissociation constant for the submicelle-calcium complex is 0.074 mM for CaCl2 and calcium phosphate buffers, vs 0.016 mM for the milk salt solution. The asymmetric frontal boundaries and higher average affinities observed with milk salts may be due to binding of calcium salts with greater affinity in addition to the binding of free Ca2+ in these solutions

  16. Laser-Casein phosphopeptide effect on remineralization of early enamel lesions in primary teeth

    OpenAIRE

    Aminabadi, Naser-Asl; Najafpour, Ebrahim; Samiei, Mohammad; Erfanparast, Leila; Anoush, Somayeh; Jamali, Zahra; Pournaghi-Azar, Fatemeh; Oskouei, Sina Ghertasi

    2015-01-01

    Background The aim of this study was to assess the effect of Nd:YAG laser irradiation following casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) application on calcium and phosphate concentration and surface microhardness (SMH) of enamel surface in artificial white spot lesions of primary teeth. Material and Methods Eighty teeth with artificial white spot lesions were randomly divided into four groups: (A) distilled and deionized water, (B) Nd:YAG laser, (C) CPP-ACP crème, & (D) CP...

  17. Casein Kinase II Phosphorylation of the Yeast Phospholipid Synthesis Transcription Factor Opi1p*

    OpenAIRE

    Chang, Yu-Fang; Carman, George M.

    2006-01-01

    The transcription factor Opi1p regulates phospholipid synthesis in the yeast Saccharomyces cerevisiae by repressing the expression of several UASINO-containing genes (e.g., INO1). Opi1p repressor activity is most active in inositol-supplemented cells. Regulation of Opi1p repressor activity is mediated by multiple phosphorylations catalyzed by protein kinases A and C. In this work, we showed that Opi1p was also phosphorylated by casein kinase II. Using purified maltose binding protein (MBP)-Op...

  18. The Bond Strength of Ceramic Brackets Bonded to Remineralized Teeth with Casein

    OpenAIRE

    Kıyak Havlucu, Sevgi; Çakırer, Banu

    2012-01-01

    The aim of this study was to evaluate the shear bond strength of ceramic brackets bonded with different adhesives to remineralized teeth with casein phosphopeptide amorphous calcium phosphate. Premolar teeth (n=120) were used in the study. Teeth were divided into 6 groups (n=20). For 60 teeth in the first 3 groups demineralization was performed before all teeth in the 6 groups were pretreated with CPP-ACP cream (GC Tooth Mousse, GC Corp., Tokyo, Japan) for 30 days. Brackets were bonded with T...

  19. The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II.

    OpenAIRE

    Meek, D W; Simon , S; Kikkawa, U; Eckhart, W

    1990-01-01

    The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kin...

  20. Mechanism of Regulation of Casein Kinase I Activity by Group I Metabotropic Glutamate Receptors

    OpenAIRE

    Liu, Feng; Virshup, David M.; Nairn, Angus C.; Greengard, Paul

    2002-01-01

    Previously, we reported that (S)-3,5-dihydroxypenylglycine (DHPG), an agonist for group I metabotropic glutamate receptors (mGluRs), stimulates CK1 and Cdk5 kinase activities in neostriatal neurons, leading to enhanced phosphorylation, respectively, of Ser-137 and Thr-75 of DARPP-32 (dopamine and cAMP-regulated phosphoprotein, 32 kDa). We have now investigated the signaling pathway that leads from mGluRs to casein kinase 1 (CK1) activation. In mouse neostriatal slices, the effect of DHPG on p...

  1. Casein kinase II phosphorylation increases the rate of serum response factor-binding site exchange.

    OpenAIRE

    Marais, R M; Hsuan, J J; McGuigan, C.; Wynne, J; Treisman, R

    1992-01-01

    Recombinant baculoviruses were used to express wild-type serum response factor (SRF) and a mutant, SRF.CKIIA, which lacks all four serine residues in the major casein kinase II (CKII) site at residues 77-90. Purified recombinant SRF binds DNA with an affinity and specificity indistinguishable from that of HeLa cell SRF, and activates transcription in vitro. Comparative phosphopeptide analysis of the wild-type and mutant proteins demonstrated that the wild-type protein is phosphorylated at the...

  2. Sequential Activation and Inactivation of Dishevelled in the Wnt/β-Catenin Pathway by Casein Kinases

    OpenAIRE

    Bernatik, Ondrej; Ganji, Ranjani Sri; Dijksterhuis, Jacomijn P.; Konik, Peter; Cervenka, Igor; Polonio, Tilman; Krejci, Pavel; Schulte, Gunnar; Bryja, Vitezslav

    2011-01-01

    Dishevelled (Dvl) is a key component in the Wnt/β-catenin signaling pathway. Dvl can multimerize to form dynamic protein aggregates, which are required for the activation of downstream signaling. Upon pathway activation by Wnts, Dvl becomes phosphorylated to yield phosphorylated and shifted (PS) Dvl. Both activation of Dvl in Wnt/β-catenin signaling and Wnt-induced PS-Dvl formation are dependent on casein kinase 1 (CK1) δ/ϵ activity. However, the overexpression of CK1 was shown to dissolve Dv...

  3. REGγ deficiency promotes premature aging via the casein kinase 1 pathway

    OpenAIRE

    Li, Lei; Zhao, Dengpan; Wei, Haibin; Yao, Liangfang; Dang, Yongyan; Amjad, Ali; Xu, Jinjin; Liu, Jiang; Guo, Linjie; Li, Dongqing; Li, Zhen; Zuo, Di; Zhang, Yuanyuan; Liu, Jian; Huang, Shixia

    2013-01-01

    Our recent studies suggest a role for the proteasome activator REG (11S regulatory particles, 28-kDa proteasome activator)γ in the regulation of tumor protein 53 (p53). However, the molecular details and in vivo biological significance of REGγ-p53 interplay remain elusive. Here, we demonstrate that REGγ-deficient mice develop premature aging phenotypes that are associated with abnormal accumulation of casein kinase (CK) 1δ and p53. Antibody array analysis led us to identify CK1δ as a direct t...

  4. Inhibition of the Casein-Kinase-1-Epsilon/Delta Prevents Relapse-Like Alcohol Drinking

    OpenAIRE

    Perreau-Lenz, Stéphanie; Vengeliene, Valentina; Noori, Hamid R.; Merlo-Pich, Emilio V.; Corsi, Mauro A; Corti, Corrado; Spanagel, Rainer

    2012-01-01

    During the past decade, it has been shown that circadian clock genes have more than a simple circadian time-keeping role. Clock genes also modulate motivational processes and have been implicated in the development of psychiatric disorders such as drug addiction. Recent studies indicate that casein-kinase 1ɛ/δ (CK1ɛ/δ)—one of the components of the circadian molecular clockwork—might be involved in the etiology of addictive behavior. The present study was initiated to study the specific role o...

  5. Effects of Whey, Caseinate, or Milk Protein Ingestion on Muscle Protein Synthesis after Exercise

    OpenAIRE

    Atsushi Kanda; Kyosuke Nakayama; Chiaki Sanbongi; Masashi Nagata; Shuji Ikegami; Hiroyuki Itoh

    2016-01-01

    Whey protein (WP) is characterized as a “fast” protein and caseinate (CA) as a “slow” protein according to their digestion and absorption rates. We hypothesized that co-ingestion of milk proteins (WP and CA) may be effective for prolonging the muscle protein synthesis response compared to either protein alone. We therefore compared the effect of ingesting milk protein (MP) to either WP or CA alone on muscle protein synthesis after exercise in rats. We also compared the effects of these milk-d...

  6. Regulation of the Drosophila melanogaster Protein, Enhancer of Rudimentary, by Casein Kinase II

    OpenAIRE

    Gelsthorpe, Mark E.; Tan, Zehui; Phillips, Anthony; Eissenberg, Joel C.; Miller, Ashley; Wallace, Janell; Tsubota, Stuart I.

    2006-01-01

    The Drosophila melanogaster gene enhancer of rudimentary, e(r), encodes a conserved protein, ER. Most ER homologs share two casein kinase II (CKII) target sites. In D. melanogaster, these sites are T18 and S24. A third CKII site, T63, has been seen only in drosophilids. The conservation of these CKII sites, particularly T18 and S24, suggests a role for these residues in the function of the protein. To test this hypothesis, these positions were mutated either to alanine as a nonphosphorylated ...

  7. Casein Kinase 2 Regulates the mRNA-destabilizing Activity of Tristetraprolin*

    OpenAIRE

    Lee, Won Hyeok; Lee, Hyun Hee; Vo, Mai-Tram; Kim, Hyo Jeong; Ko, Myoung Seok; Im, Yeong-Cheol; Min, Young Joo; Lee, Byung Ju; Cho, Wha Ja; Park, Jeong Woo

    2011-01-01

    Tristetraprolin (TTP) is an AU-rich element-binding protein that regulates mRNA stability. We previously showed that TTP acts as a negative regulator of VEGF gene expression in colon cancer cells. The p38 MAPK pathway is known to suppress the TTP activity. However, until now the signaling pathway to enhance TTP function is not well known. Here, we show that casein kinase 2 (CK2) enhances the TTP function in the regulation of the VEGF expression in colon cancer cells. CK2 increased TTP protein...

  8. Specificity of an extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine beta-casein.

    OpenAIRE

    Rattray, F P; Fox, P. F.; Healy, A.

    1997-01-01

    The specificity of the extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine beta-casein was studied. Hydrolysis was monitored over time by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and urea-PAGE. The major pH 4.6-soluble peptides were isolated by high-performance liquid chromatography and identified by N-terminal amino acid sequencing and mass spectrometry. The major sites of hydrolysis were Ser-18-Ser-19, Glu-20-Glu-21, Gln-56-Ser-57, Gln-72-Asn-73, ...

  9. Specificity of an extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine alpha s1-casein.

    OpenAIRE

    Rattray, F P; Fox, P. F.; Healy, A.

    1996-01-01

    The specificity of the extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine alpha s1-casein was studied. Hydrolysis was monitored over time by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and urea-PAGE. The major pH 4.6-soluble peptides were isolated by high-performance liquid chromatography and identified by N-terminal amino acid sequencing and mass spectrometry. The time course of peptide formation indicated that His-8-Gln-9, Ser-161-Gly-162, and eithe...

  10. Radioprotective properties of casein phospho peptides (CPP) in gamma irradiated animal models

    International Nuclear Information System (INIS)

    In search of radioprotective food supplement the antigenotoxic role of casein phospho peptides (CPP) from fermented milk were tested in animal models using fish and Swiss albino mice. The antigenotoxic property was assayed by comparing the number of micronuclei, binucleated and multinucleated erythrocytes formed in the unfed control mice and fish with the CPP fed test mice and fish cells. The antigenotoxic property of CPP proved of its radioprotective nature. This potential can be harnessed to develop a new class of radioprotective nutraceuticals which would be of immense help as food supplement for workers working with low background radiation and who are in constant exposure to natural radiation. (author)

  11. Casein Phosphopeptide-Amorphous Calcium Phosphate and Shear Bond Strength of Adhesives to Primary Teeth Enamel

    OpenAIRE

    Farokh Gisovar, Elham; Hedayati, Nassim; Shadman, Niloofar; Shafiee, Leila

    2015-01-01

    Background: CPP-ACP (Phosphopeptide-Amorphous Calcium Phosphate) has an important role in caries prevention in pediatric patients. This study was done, because of the great use of CPP-ACP and the need for restoration for teeth treated with CPP-ACP as well as the importance of shear bond strength of adhesives in the success of restorations. Objectives: This study aimed to evaluate the effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on shear bond strength of dental adhesiv...

  12. Amino acid sequence of the beta subunit of bovine lung casein kinase II.

    OpenAIRE

    Takio, K.; Kuenzel, E A; Walsh, K. A.; Krebs, E G

    1987-01-01

    The amino acid sequence of the 209-residue beta subunit of bovine lung casein kinase II has been determined. Excluding the amino-terminal blocking group, which was not identified, the molecular weight of the polypeptide chain is 24,239. A marked polarity of the beta subunit is indicated by clusters of negative charges in the amino-terminal region and of positive charges in the carboxyl-terminal region. Whereas the beta subunit shows no homology with any known protein, a segment of the sequenc...

  13. Use of cold microfiltration to produce unique ${\\beta}$-casein enriched milk gels

    OpenAIRE

    Van Hekken, Diane; Holsinger, Virginia

    2000-01-01

    Bovine milk was modified using cold microfiltration to produce $\\beta$-casein enriched fractions with unique gelation properties. Skim milk at $4 ^\\circ$C was microfiltered using membranes with pore diameter of $0.2 \\mu$m, $0.1 \\mu$m, or 100 000 g.mol$^{-1}$ MWCO. Resulting permeates were filtered again using membranes with 10 000 g.mol$^{-1}$ MWCO to create retentates with 9% solids that were used in the gelation study. Ash, calcium, lactose, solids, and protein contents of all fractions wer...

  14. The casein kinase II beta subunit binds to Mos and inhibits Mos activity.

    OpenAIRE

    Chen, M.; D. Li; Krebs, E G; Cooper, J. A.

    1997-01-01

    Mos is a germ cell-specific serine/threonine kinase and is required for Xenopus oocyte maturation. Active Mos stimulates a mitogen-activated protein kinase (MAPK) by directly phosphorylating and activating MAPK kinase (MKK). We report here that the Xenopus homolog of the beta subunit of casein kinase II (CKII beta) binds to and regulates Mos. The Mos-interacting region of CKII beta was mapped to the C terminus. Mos bound to CKII beta in somatic cells ectopically expressing Mos and CKII beta a...

  15. IDENTIFICATION OF BETA-LACTOGLOBULIN AND KAPPA-CASEIN GENOTYPES IN CATTLE

    OpenAIRE

    C.D. TESIO; DINISCHIOTU ANCA; MANEA MARIA ADINA; GEORGESCU S. E.; R.A. VĂTĂŞESCU-BALCAN; COSTACHE MARIETA

    2007-01-01

    Beta-lactoglobulin (b-Lg) and kappa-casein (k-Cn) are two of the most importantproteins in the mammals’ milk synthesized by the epithelial cells of the mammaryglands. They play a crucial role in the milk quality and coagulation process(production of cheese and butter). The PCR-RFLP test was performed to distinguishthe different alleles in a population of Romanian Black Spotted cattle, a dairy breed.Genetic polymorphism was detected by digestion with the endonucleases Hae III (b-Lg) and Hinf I...

  16. Casein and soya-bean protein have different effects on whole body protein turnover at the same nitrogen balance

    DEFF Research Database (Denmark)

    Nielsen, K; Kondrup, J; Elsner, Petteri;

    1994-01-01

    The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or...... recovered from urinary ammonia and urea during isotope steady state for measurement of protein synthesis and protein degradation. Compared with starvation the protein-free diet decreased N excretion by 75%, probably by increasing the rate of reutilization of amino acids from endogenous proteins for protein...... synthesis. The protein diets produced a positive N balance which was independent of the protein source. Intact and hydrolysed casein increased protein synthesis 2.6- and 2.0-fold respectively, compared with the protein-free diet. Protein degradation increased 1.4- and 1.2-fold respectively. Hydrolysed soya...

  17. Casein and soya-bean protein have different effects on whole body protein turnover at the same nitrogen balance

    DEFF Research Database (Denmark)

    Nielsen, K; Kondrup, J; Elsner, Petteri;

    1994-01-01

    The present study examined whether different proteins have different effects on whole-body protein turnover in adult rats. The rats were either starved, given a protein-free but energy-sufficient diet (1 MJ/kg body weight (BW) per d) or a diet containing intact casein, hydrolysed casein, or...... recovered from urinary ammonia and urea during isotope steady state for measurement of protein synthesis and protein degradation. Compared with starvation the protein-free diet decreased N excretion by 75%, probably by increasing the rate of reutilization of amino acids from endogenous proteins for protein......-bean protein did not increase protein synthesis but decreased protein degradation by 35% compared with the protein-free diet. Compared with the hydrolysed soya-bean protein, intact casein resulted in 2.2- and 2.8-fold higher rates of protein synthesis and degradation respectively. These results are not easily...

  18. In vitro digestibility of beta-casein and beta-lactoglobulin under simulated human gastric and duodenal conditions: A multi-laboratory evaluation

    DEFF Research Database (Denmark)

    Mandalari, G.; Adel-Patient, K.; Barkholt, Vibeke;

    2009-01-01

    gastroduodenal digestion in the presence or absence of phosphatidylcholine (PC). Both beta-casein and beta-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of beta-casein digestion in the low-protease assay were slower, beta-Lg being pepsin...

  19. Modification of Casein and its Application in Leather Finishing%酪素的改性方法及其在皮革涂饰中的应用

    Institute of Scientific and Technical Information of China (English)

    唐瑶; 付柯; 赵丹; 张哲

    2015-01-01

    简述了酪素的性质,综述了酪素的改性方法及在皮革涂饰中的应用。%The properties of casein were briefly introduced. Modification methods of casein and its applica-tion in synthesizing leather finishing agents were reviewed.

  20. Chaperone-like activity of β-casein and its effect on residual in vitro activity of horseradish peroxidase

    DEFF Research Database (Denmark)

    Sulewska, Anna Maria; Olsen, Karsten; Sørensen, Jens Christian; Øgendal, Lars Holm

    2014-01-01

    In this study, the residual activity horseradish peroxidase was used as a novel marker of chaperone-like activity of β-casein under elevated temperature. It was shown that β-casein does affect residual activity of horseradish peroxidase (HRP) depending on the concentration and molar ratio between...

  1. Alpha S1-casein polymorphisms in camel (Camelus dromedarius) and descriptions of biological active peptides and allergenic epitopes.

    Science.gov (United States)

    Erhardt, Georg; Shuiep, El Tahir Salih; Lisson, Maria; Weimann, Christina; Wang, Zhaoxin; El Zubeir, Ibtisam El Yas Mohamed; Pauciullo, Alfredo

    2016-06-01

    Milk samples of 193 camels (Camelus dromedarius) from different regions of Sudan were screened for casein variability by isoelectric focusing. Kappa-casein and beta-casein were monomorphic, whereas three protein patterns named αs1-casein A, C, and D were identified. The major allele A revealed frequencies of 0.79 (Lahaoi), 0.75 (Shanbali), 0.90 (Arabi Khali), and 0.88 (Arabi Gharbawi) in the different ecotypes. CSN1S1*C shows a single G > T nucleotide substitution in the exon 5, leading to a non-synonymous amino acid exchange (p.Glu30 > Asp30) in comparison to CSN1S1*A and D. At cDNA level, no further single nucleotide polymorphisms could be identified in CSN1S1* A, C, and D, whereas the variants CSN1S1*A and CSN1S1*C are characterized by missing of exon 18 compared to the already described CSN1S1*B, as consequence of DNA insertion of 11 bp at intron 17 which alter the pre-mRNA spliceosome machinery. A polymerase chain-restriction fragment length polymorphism method (PCR-RFLP) was established to type for G > T nucleotide substitution at genomic DNA level. The occurrence and differences of IgE-binding epitopes and bioactive peptides between αs1-casein A, C, and D after digestion were analyzed in silico. The amino acid substitutions and deletion affected the arising peptide pattern and thus modifications between IgE-binding epitopes and bioactive peptides of the variants were found. The allergenic potential of these different peptides will be investigated by microarray immunoassay using sera from milk-sensitized individuals, as it was already demonstrated for bovine αs1-casein variants. PMID:26922739

  2. THE INFLUENCE OF K-CASEIN ALLELES ON MILK PRODUCTION AND QUALITY IN A HOLSTEIN-FRISIAN COW POPULATION

    Directory of Open Access Journals (Sweden)

    I. BENCSIK

    2013-12-01

    Full Text Available Milk production and its composition are determined by quantitative loci, whichunder the influence of some environmental factors are producing an allelicvariability, meaning a genetic polymorphism of the gene. K-casein is a milk proteinwhose genetic polymorphism can serve as molecular marker for milk production,composition and industrial processing suitability. The allelic variants for k-casein Aand B are the most common and the most important of them. The experiments wereconducted on 24 Holstein-Friesian milking cows from a private farm in Giroc. Themilk production on a normal lactation is 8444 milk kg/305 days, with a fat percentof 3.9 and a protein percent of 3.3. The cows were divided in three groups AA, ABand BB in function of the genotyped obtained after the allelic variantsdetermination. The DNA isolation was made from hair roots and blood, the cowpopulation studied is not in genetic equilibrium fore k-casein gene, the frequency ofallele A is 0.43 and the frequency of B allele is 0.58. The highest genotype frequencywas 0.5 for CSN3-AB genotype, the BB genotype had 0.33 frequency, and the lowestfrequency was 0.17 for AA genotype. The mean daily milk production from cowswith BB genotype for k-casein is significant (p<5% higher compared to the allelicvariant AA. The fat percent is significant higher at the allelic variant AA comparedto the other allelic variants (AB and BB of the k-casein gene. Between the fatpercent of the three genotypes variants of K-casein (AA, AB and BB there are nosignificant differences.

  3. THE INFLUENCE OF K-CASEIN ALLELES ON MILK PRODUCTION AND QUALITY IN A HOLSTEIN-FRISIAN COW POPULATION

    Directory of Open Access Journals (Sweden)

    BENCSIK I.

    2007-01-01

    Full Text Available Milk production and its composition are determined by quantitative loci, whichunder the influence of some environmental factors are producing an allelicvariability, meaning a genetic polymorphism of the gene. K-casein is a milk proteinwhose genetic polymorphism can serve as molecular marker for milk production,composition and industrial processing suitability. The allelic variants for k-casein Aand B are the most common and the most important of them. The experiments wereconducted on 24 Holstein-Friesian milking cows from a private farm in Giroc. Themilk production on a normal lactation is 8444 milk kg/305 days, with a fat percentof 3.9 and a protein percent of 3.3. The cows were divided in three groups AA, ABand BB in function of the genotyped obtained after the allelic variantsdetermination. The DNA isolation was made from hair roots and blood, the cowpopulation studied is not in genetic equilibrium fore k-casein gene, the frequency ofallele A is 0.43 and the frequency of B allele is 0.58. The highest genotype frequencywas 0.5 for CSN3-AB genotype, the BB genotype had 0.33 frequency, and the lowestfrequency was 0.17 for AA genotype. The mean daily milk production from cowswith BB genotype for k-casein is significant (p<5% higher compared to the allelicvariant AA. The fat percent is significant higher at the allelic variant AA comparedto the other allelic variants (AB and BB of the k-casein gene. Between the fatpercent of the three genotypes variants of K-casein (AA, AB and BB there are nosignificant differences.

  4. Rennet-induced gelation of concentrated milk in the presence of sodium caseinate: differences between milk concentration using ultrafiltration and osmotic stressing.

    Science.gov (United States)

    Krishnankutty Nair, P; Corredig, M

    2015-01-01

    Concentrating milk is a common unit operation in the dairy industry. With the reduction of water, the particles interact more frequently with each other and the functionality of the casein micelles may depend on the interactions occurring during concentration. The objective of this research was to investigate the effect of concentration on the renneting properties of the casein micelles by comparing 2 concentration methods: ultrafiltration and osmotic stressing. Both methods selectively concentrate the protein fraction of milk, while the composition of the soluble phase is unaltered. To evaluate possible differences in the rearrangements of the casein micelles during concentration, renneting properties were evaluated with or without the addition of soluble caseins, added either before or after concentration. The results indicate that casein micelles undergo rearrangements during concentration and that shear during membrane filtration may play a role in affecting the final properties of the milk. PMID:25468692

  5. Genetic Variants and Allele Frequencies of Kappa Casein in Egyptian Cattle and Buffalo Using PCR-RFLP

    OpenAIRE

    Eman M. Gouda; Mona Kh. Galal; Samy A. Abdelaziz

    2013-01-01

    Kappa casein (K-Ca) genetic variations affected quality and composition of the milk. Several variants of Kappa casein (K-Ca) gene locus IV have been reported with special interest for the ‘B’ allele for its relation to the milk protein and fat yields. Genotyping and allelic frequencies of K-Ca among Native Egyptian breeds of cattle and buffalo were the aim of the present study. PCR amplification of DNA isolated from 300 blood samples collected from Holstein and Baladi cattle and buffalo were ...

  6. The usefulness of casein-specific IgE and IgG4 antibodies in cow's milk allergic children

    Directory of Open Access Journals (Sweden)

    Ito Komei

    2012-01-01

    Full Text Available Abstract Background Cow's milk allergy is one of the most common food allergies among younger children. We investigated IgE antibodies to milk, and IgE and IgG4 antibodies to casein, α-lactalbumin and β-lactoglobulin in cow's milk allergic (CMA and non-allergic (non-CMA children in order to study their clinical usefulness. Methods Eighty-three children with suspected milk allergy (median age: 3.5 years, range: 0.8-15.8 years were diagnosed as CMA (n = 61 or non-CMA (n = 22 based on an open milk challenge or convincing clinical history. Their serum concentrations of allergen-specific (s IgE and IgG4 antibodies were measured using ImmunoCAP®. For the sIgG4 analysis, 28 atopic and 31 non-atopic control children were additionally included (all non-milk sensitized. Results The CMA group had significantly higher levels of milk-, casein- and β-lactoglobulin-sIgE antibodies as compared to the non-CMA group. The casein test showed the best discriminating performance with a clinical decision point of 6.6 kUA/L corresponding to 100% specificity. All but one of the CMA children aged > 5 years had casein-sIgE levels > 6.6 kUA/L. The non-CMA group had significantly higher sIgG4 levels against all three milk allergens compared to the CMA group. This was most pronounced for casein-sIgG4 in non-CMA children without history of previous milk allergy. These children had significantly higher casein-sIgG4 levels compared to any other group, including the non-milk sensitized control children. Conclusions High levels of casein-sIgE antibodies are strongly associated with milk allergy in children and might be associated with prolonged allergy. Elevated casein-sIgG4 levels in milk-sensitized individuals on normal diet indicate a modified Th2 response. However, the protective role of IgG4 antibodies in milk allergy is unclear.

  7. Comparison of Casein Micelles Micrographs in Raw and Pasteurized Skim Milk in Different pHs by SEM and TEM

    OpenAIRE

    H. Lamea; M. R. Ehsani; H. Ezzatpanah

    2005-01-01

    In this research some properties of the casein micelles in the raw and pasteurized milk were studied by electron microscopy. SEM and TEM were used to evaluate the differences in acidified casein micelles of raw and pasteurized milk at the Iso Electric Point (pH=4.6). Milk samples were taken from research pilot plant of The College of Agriculture. Milk was pasteurized by the L.T.L.T. method in the same pilot plant. The samples of raw and pasteurized milk were divided into two parts. One part o...

  8. Postprandial response of plasma IL-6 to isoenergetic meals rich in casein or potato singly and combined in obese women

    OpenAIRE

    Patrick J Manning; Sutherland, Wayne H. F.; de Jong, Sylvia A.; Ryalls, Anne R.; Berry, Elizabeth A.

    2013-01-01

    Milk consumption decreases inflammatory stress in overweight and obese subjects. Casein is the major protein in milk and enhances the secretion of insulin that has anti-inflammatory activity. The aim of the present study was to compare the acute effect of meals rich in casein and carbohydrate and a combination of both nutrients on postprandial plasma concentrations of IL-6, a marker of inflammation, in obese women. A total of twenty-five obese women aged 38–68 years consumed isoenergetic meal...

  9. The Schizosaccharomyces pombe casein kinase II alpha and beta subunits: evolutionary conservation and positive role of the beta subunit.

    OpenAIRE

    Roussou, I; Draetta, G

    1994-01-01

    Casein kinase II is a key regulatory enzyme involved in many cellular processes, including the control of growth and cell division. We report the molecular cloning and sequencing of cDNAs encoding the alpha and the beta subunits of casein kinase II of Schizosaccharomyces pombe. The deduced amino acid sequence of Cka1, the alpha catalytic subunit, shows high sequence similarity to alpha subunits identified in other species. The amino acid sequence of Ckb1, the S. pombe beta subunit, is 57% ide...

  10. Coagulation properties and Nostrano di Primiero cheese yield of milk from Brown grazing cows of different k-casein genotype

    Directory of Open Access Journals (Sweden)

    G. Gaiarin

    2011-03-01

    Full Text Available Genetic variants of milk proteins affect composition, technological characteristics and, as a consequence, cheesemaking properties of milk (Jakob and Puhan, 1992; Mariani and Summer, 1999. The effects of αS1, β and k-caseins variants, on milk coagulation properties are well known (Grosclaude, 1988. Cheesemaking involving many cheese varieties showed shorter clotting and curd firming times, higher curd firmness and cheese yields with the k-casein BB variant (Van Den Berg et al., 1992;Walsh et al., 1998......

  11. Cytotoxicity assessment of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP paste

    Directory of Open Access Journals (Sweden)

    Sandra Kalil Bussadori

    2010-01-01

    Full Text Available Introduction: Casein phosphopeptides (CPP have been shown to be good carriers of calcium, phosphate, and hydroxide ions to promote enamel remineralization with applications in oral care products, professional dental products, and food products. Objectives: Evaluate the cytotoxicity of a casein phosphopeptideamorphous calcium phosphate (CPP-ACP paste in rat fibroblasts. Materials and methods: Cytotoxicity was measured by the Trypan blue dye exclusion assay and the MTT assay. Results: Long term (1, 3, 5, and 7 days and short term tests (0, 4, 8, and 12 hours were performed with the Trypan blue dye exclusion assay. Data were analyzed with ANOVA and Kruskall Wallis tests. After one day, the cell viability of the CPP-ACP study group was between 80 and 100%. In the short term experiments, a reduction of cell viability in the study group was observed compared with the control group; yet viable cells were present over the evaluation period. With the MTT assay, a viability of 76.43% was observed in the study group, a significant difference from the control group (P = 0.003. Conclusion: CPP-ACP paste demonstrates low cytotoxicity in rat fibroblast culture.

  12. Novel hierarchical microparticles super-assembled from nanoparticles with the induction of casein micelles

    Energy Technology Data Exchange (ETDEWEB)

    Xiong, Xiaopeng, E-mail: xpxiong@xmu.edu.cn; Duan, Jiangjiang; Wang, Yong; Yu, Zhaoju [Xiamen University, Department of Materials Science and Engineering, College of Materials (China)

    2013-08-15

    We have demonstrated a solution-based synthesis of novel waxberry-like hierarchical ZnO microparticles in the presence casein micelles under mild conditions. The microstructures of the sub-micrometer-sized hierarchical microparticles were characterized, and the synthesis conditions were optimized. The formation mechanism of the hierarchical microparticle was analyzed through control experiments. The hierarchical ZnO microparticles are found to be super-assemblies of 30-70 nm ZnO nanoparticles, which are thought to be based on casein micelle induction followed by Ostwald ripening. In the same manner, copper-based hierarchical microparticles with a similar morphology have also been successfully synthesized. By controlling the synthetic time or temperature, solid or hollow microparticles can be fabricated. The narrowly distributed ZnO microparticles have a high specific surface area, exhibiting great potential application in fields such as photocatalytic and energy conversion. Our findings may meanwhile open a new bottom-up strategy in order to construct structurally sophisticated nanomaterials.

  13. Dystroglycan loss disrupts polarity and beta-casein induction inmammary epithelial cells by perturbing laminin anchoring

    Energy Technology Data Exchange (ETDEWEB)

    Weir, M. Lynn; Oppizzi, Maria Luisa; Henry, Michael D.; Onishi,Akiko; Campbell, Kevin P.; Bissell, Mina J.; Muschler, John L.

    2006-02-17

    Precise contact between epithelial cells and their underlying basement membrane is critical to the maintenance of tissue architecture and function. To understand the role that the laminin receptor dystroglycan (DG) plays in these processes, we assayed cell responses to laminin-111 following conditional ablation of DG expression in cultured mammary epithelial cells (MECs). Strikingly, DG loss disrupted laminin-111-induced polarity and {beta}-casein production, and abolished laminin assembly at the step of laminin binding to the cell surface. DG re-expression restored these deficiencies. Investigations of mechanism revealed that DG cytoplasmic sequences were not necessary for laminin assembly and signaling, and only when the entire mucin domain of extracellular DG was deleted did laminin assembly not occur. These results demonstrate that DG is essential as a laminin-111 co-receptor in MECs that functions by mediating laminin anchoring to the cell surface, a process that allows laminin polymerization, tissue polarity, and {beta}-casein induction. The observed loss of laminin-111 assembly and signaling in DG-/-MECs provides insights into the signaling changes occurring in breast carcinomas and other cancers, where DG's laminin-binding function is frequently defective.

  14. Supplemental barley protein and casein similarly affect serum lipids in hypercholesterolemic women and men.

    Science.gov (United States)

    Jenkins, David J A; Srichaikul, Korbua; Wong, Julia M W; Kendall, Cyril W C; Bashyam, Balachandran; Vidgen, Edward; Lamarche, Benoicirct; Rao, A Venketeshwer; Jones, Peter J H; Josse, Robert G; Jackson, Chung-Ja C; Ng, Vivian; Leong, Tracy; Leiter, Lawrence A

    2010-09-01

    High-protein diets have been advocated for weight loss and the treatment of diabetes. Yet animal protein sources are often high in saturated fat and cholesterol. Vegetable protein sources, by contrast, are low in saturated fat and without associated cholesterol. We have therefore assessed the effect on serum lipids of raising the protein intake by 5% using a cereal protein, barley protein, as part of a standard therapeutic diet. Twenty-three hypercholesterolemic men and postmenopausal women completed a randomized crossover study comparing a bread enriched with either barley protein or calcium caseinate [30 g protein, 8374 kJ (2000 kcal)] taken separately as two 1-mo treatment phases with a minimum 2-wk washout. Body weight and diet history were collected weekly during each treatment. Fasting blood samples were obtained at wk 0, 2, and 4. Palatability, satiety, and compliance were similar for both the barley protein- and casein-enriched breads, with no differences between the treatments in effects on serum LDL cholesterol or C-reactive protein, measures of oxidative stress, or blood pressure. Nevertheless, because no adverse effects were observed on cardiovascular risk factors, barley protein remains an additional option for raising the protein content of the diet. PMID:20668250

  15. Emulsifying properties of maillard conjugates produced from sodium caseinate and locust bean gum

    Directory of Open Access Journals (Sweden)

    F. A. Perrechil

    2014-06-01

    Full Text Available Emulsifying properties of sodium caseinate -locust bean gum Maillard conjugates produced at different temperatures (54 - 96 ºC, protein/polysaccharide ratios (0.3 - 1.0 and reaction times (1 - 24 hours were evaluated. Conjugate formation was confirmed by formation of color and high molecular weight fractions and the decrease of the αs- and β-casein bands. The emulsions stabilized by Maillard conjugates showed good stability. The mean droplet diameter (d32 tended to decrease with the increase of incubation time and temperature, except at extreme conditions (24 hours and 90 ºC or 96 ºC when the partial degradation of the conjugates was probably favored, resulting in phase separation of emulsions. The emulsion viscosity decreased with the increase in the protein/polysaccharide ratio and with the degradation of the conjugates. The conditions used in the experimental design made the optimization of the conjugate production viable, which showed greater emulsifier properties than the pure protein under acid conditions.

  16. Isolation of prolyl endopeptidase inhibitory peptides from a sodium caseinate hydrolysate.

    Science.gov (United States)

    Hsieh, Cheng-Hong; Wang, Tzu-Yuan; Hung, Chuan-Chuan; Hsieh, You-Liang; Hsu, Kuo-Chiang

    2016-01-01

    Prolyl endopeptidase (PEP) has been associated with neurodegenerative disorders, and the PEP inhibitors can restore the memory loss caused by amnesic compounds. In this study, we investigated the PEP inhibitory activity of the enzymatic hydrolysates from various food protein sources, and isolated and identified the PEP inhibitory peptides. The hydrolysate obtained from sodium caseinate using bromelain (SC/BML) displayed the highest inhibitory activity of 86.8% at 5 mg mL(-1) in the present study, and its IC50 value against PEP was 0.77 mg mL(-1). The F-5 fraction by RP-HPLC (reversed-phase high performance liquid chromatography) from SC/BML showed the highest PEP inhibition rate of 88.4%, and 9 peptide sequences were identified. The synthetic peptides (1245.63-1787.94 Da) showed dose-dependent inhibition effects on PEP as competitive inhibitors with IC50 values between 29.8 and 650.5 μM. The results suggest that the peptides derived from sodium caseinate have the potential to be PEP inhibitors. PMID:26574880

  17. Formation of sterilized edible-films based on caseinates: Effects of calcium and plasticizers

    International Nuclear Information System (INIS)

    Gamma-irradiation was used to produce free-standing sterilized edible films based on caseinate. The effect of calcium ions (Ca2+) and two plasticizers, namely propylene glycol (PG) and triethylene glycol (TEG) were investigated, as well as the effect of the irradiation on both the gel formation and mechanical properties of the resulting films. Gamma-irradiation provoked formation of bityrosine, i.e. crosslinks, accounting for the increase of the puncture strength of films. The presence of PG or TEG enhanced the formation of crosslinks, leading to an improved mechanical strength of films. TEG was found to interact more favorably with the caseinate than PG, being responsible for the improved film extensibility. Addition of CA2+ caused the formation of gels. The breaking strength of gels was directly related to the concentration of Ca2+, while the puncture strength of films was found to be almost independent. Moreover, high irradiation dose seemed to affect the protein structure, accounting for the decrease of the breaking strength of gels and for the depreciation of the mechanical behavior of films

  18. Radiotracer study of phosphate exchange between whey and casein micelles in cow's milk

    International Nuclear Information System (INIS)

    Radiotracer method has been applied to study exchange of calcium ions between the whey calcium salts and micellar calcium phosphate (MCP). The present paper deals with a similar study pertaining to phosphate ions. 32P-labelled Na2HPO4 was used as the radiotracer for inorganic phosphates of milk. After addition of the radiotracer to skimmed-milk, samples were taken regularly for 700 hours. In the samples casein micelles were separated from whey by ultracentrifugation and finally the radiotracer quantity i.e. 32P-concentration in the whey samples was measured using a Liquid Scintillation Counter. Compartmental analysis and modelling were used to evaluate the thus obtained time curves for radiotracer quantity in whey. This analysis revealed the presence of three phosphate compartments i.e. exchangeable phosphate entities; one being the whey phosphate. The other two are associated with the exchangeable phosphates of MCP. The mean residence times of phosphate in the latter two compartment differ considerably pointing at two distinctly different embeddings of phosphate groups in the structure of the micellar calcium phosphate of the cow's milk casein. The obtained results are in fair agreement with the mentioned model of MCP

  19. Optimization of production of Brucella abortus S19 culture in bioreactor using soyabean casein digest medium.

    Science.gov (United States)

    Kamaraj, Govindasamy; Rajendra, Lingala; Shankar, Chinchkar Ramachandra; Srinivasan, Villuppanoor Alwar

    2010-10-01

    A method of cultivating Brucella abortus S19 culture in bioreactor was attempted using three different media. Culture conditions in bioreactor were optimized by varying agitation and aeration parameters. Varying the aeration ranging from 0.5 vvm to 0.8 vvm and agitation rate ranging from 250 rpm to 400 rpm during bacterial growth was found to yield highest viable count within 48 hours of culture period. A count of > 1 x 10(11) CFU per ml within 48 to 60 hours post seeding was obtained consistently in all five consecutive batches (P > 0.05) with 6 x 10(11) CFU per ml being the maximum yield when the organism is grown in soyabean casein digest medium. B. abortus S19 maintained its smooth characteristics throughout its growth in bioreactor. The vaccine prepared with soyabean casein digest medium was found to be potent and safe with a protective index of 3.33 in mice. The vaccine was tested in 10 cattle calves of 3 to 13 months age and all the vaccinated animals were seropositive on 28, 60, 90, 120 and 150 days post-vaccination when analyzed by fluorescence polarization assay (FPA). PMID:21213590

  20. Nutritional management of (some) autism: a case for gluten- and casein-free diets?

    Science.gov (United States)

    Whiteley, Paul

    2015-08-01

    Autism spectrum disorders represent a diverse and heterogeneous array of conditions unified by the variable presence of specific behaviours impacting social and communicative functions (social affect) alongside other presentation. Common overt characteristics may come about as a consequence of several different genetic and biological processes differentially manifesting across different people or groups. The concept of plural 'autisms' is evolving, strengthened by an increasingly important evidence base detailing different developmental trajectories across the autism spectrum and the appearance of comorbidity variably interacting with core symptoms and onwards influencing quality of life. Reports that dietary intervention, specifically the removal of foods containing gluten and/or casein from the diet, may impact on the presentation of autism for some, complement this plural view of autism. Evidence suggestive of differing responses to the use of a gluten- and casein-free diet, defined as best- and non-response, has combined with some progress on determining the underlying genetic and biological correlates potentially related to such dietary elements. The preliminary suggestion of a possible diet-related autism phenotype is the result. This review will highlight several pertinent aspects onwards to an effect of food in some cases of autism including research on the pharmacological activity of food metabolites, immune response, issues with gut barrier function and some contribution from the gut microbiota. These represent promising areas in need of far greater research inspection in order to potentially define such a diet-related subgroup on the autism spectrum. PMID:25311313

  1. Impact of cationic surfactant on the self-assembly of sodium caseinate.

    Science.gov (United States)

    Vinceković, Marko; Curlin, Marija; Jurašin, Darija

    2014-08-27

    The impact of a cationic surfactant, dodecylammonium chloride (DDACl), on the self-assembly of sodium caseinate (SC) has been investigated by light scattering, zeta potential, and rheological measurements as well as by microscopy (transmission electron and confocal laser scanning microscopy). In SC dilute solutions concentration-dependent self-assembly proceeds through the formation of spherical associates and their aggregation into elongated structures composed of connected spheres. DDACl interacts with SC via its hydrophilic and hydrophobic groups, inducing changes in SC self-assembled structures. These changes strongly depend on the surfactant aggregation states (monomeric or micellar) as well as concentration ratio of both components, leading to the formation of soluble and insoluble complexes of nano- to microdimensions. DDACl monomers interact with SC self-assembled entities in a different way compared to their micelles. Surfactant monomers form soluble complexes (similar to surfactant mixed micelles) at lower SC concentration but insoluble gelatinous complexes at higher SC concentration. At surfactant micellar concentration soluble complexes with casein chains wrapped around surfactant micelles are formed. This study suggests that the use of proper cationic surfactant concentration will allow modification and control of structural changes of SC self-assembled entities. PMID:25078419

  2. Novel hierarchical microparticles super-assembled from nanoparticles with the induction of casein micelles

    International Nuclear Information System (INIS)

    We have demonstrated a solution-based synthesis of novel waxberry-like hierarchical ZnO microparticles in the presence casein micelles under mild conditions. The microstructures of the sub-micrometer-sized hierarchical microparticles were characterized, and the synthesis conditions were optimized. The formation mechanism of the hierarchical microparticle was analyzed through control experiments. The hierarchical ZnO microparticles are found to be super-assemblies of 30–70 nm ZnO nanoparticles, which are thought to be based on casein micelle induction followed by Ostwald ripening. In the same manner, copper-based hierarchical microparticles with a similar morphology have also been successfully synthesized. By controlling the synthetic time or temperature, solid or hollow microparticles can be fabricated. The narrowly distributed ZnO microparticles have a high specific surface area, exhibiting great potential application in fields such as photocatalytic and energy conversion. Our findings may meanwhile open a new bottom-up strategy in order to construct structurally sophisticated nanomaterials

  3. Evaluation of the hydrophilic behaviour of a β-casein peptide by molecular dynamics simulation

    Science.gov (United States)

    Wang, C. X.; Douillard, R.; Tran, V.

    1994-12-01

    Molecular dynamics simulations of a beta-casein peptide in the alpha-helix conformation in vacuum and aqueous solution have been performed to study the hydrophilic (and conversely hydrophobic) and hydration behavior of this peptide. Both simulations were performed for a duration of 150 ps at 300 K using the GROMOS force field, with apolar hydrogen atoms treated as united atoms, and the SPC water model. A general conclusion is that the beta-casein peptide hydrophilic behaviour can be characterized by comparison of (solvent) accessible surface area (SASA) and of the radius of gyration (R(sub g)) in vacuum and in aqueous solution. These two criterion values increase when the peptide is moved from vacuum to water. For each amino-acid, the SASA difference gives a picture of their hydrophobicity which compares favorably with the standard scale used to predict the hydrophobicity profile of the polypeptide. The hydrogen bonding analysis has been carried out and the result of peptide-water hydrogen bonding has been satisfactory compared with previous simulation and nuclear magnetic resonance NMR experiment. From calculations of the diffusion coefficient of water around the different atom types of peptide, it is found that the diffusion coefficient of water near polar atom is the smallest one, which is consistent with the recent experimental and simulation data.

  4. Genetic polymorphism of β-lactoglobulin and κ-casein of cattle breeds in Croatia

    Directory of Open Access Journals (Sweden)

    Ante Ivanković

    2011-12-01

    Full Text Available Profitable milk production respects the interests of producers, processing industries, consumer requirements and welfare of animals. Development of new methods of direct gene analysis responsible for milk proteins polymorphism provide new tools to raise the profitability of milk production and dairy products through implementation of breed genetic profile in breeding program. Because of necessity to determinate genetic profiles of cattle breeds in Croatia using new analytical methods, the ratio of dominant allelic polymorphic variants of beta-lactoglobulin (β-Lg and kappa-casein (κ-CN is defined. The share of beta-lactoglobulin B variant is dominant in all investigated cattle breeds (>52.9 %. Kappa- casein allelic variant A is dominant in selected cattle breeds (60.7-76.4 %, while the share of B variant is significantly more presented in autochthonous cattle breeds (48.2-84.1 %. Knowledge about genetic profile of breeds due to studied polymorphic variants of milk proteins is useful in further breeding development and economic reaffirmation of cattle breeds, especially autochthonous ones.

  5. Phosphorylation of αS1-casein is regulated by different genes.

    Science.gov (United States)

    Bijl, E; van Valenberg, H J F; Huppertz, T; van Hooijdonk, A C M; Bovenhuis, H

    2014-11-01

    Casein phosphorylation is a posttranslational modification catalyzed by kinase enzymes that attach phosphate groups to specific AA in the protein sequence. This modification is one of the key factors responsible for the stabilization of calcium phosphate nanoclusters in casein micelles and for the internal structure of the casein micelles. α(S1)-Casein (α(s1)-CN) is of special interest because it constitutes up to 40% of the total casein fraction in milk, and it has 2 common phosphorylation states, with 8 (α(S1)-CN-8P) and 9 (α(S1)-CN-9P) phosphorylated serine residues. Factors affecting this variation in the degree of phosphorylation are not currently known. The objective of this research was to determine the genetic background of α(S1)-CN-8P and α(S1)-CN-9P. The genetic and phenotypic correlation between α(S1)-CN-8P and α(S1)-CN-9P was low (0.18 and 0.19, respectively). This low genetic correlation suggests a different genetic background. These differences were further investigated by means of a genome-wide association study, which showed that both α(S1)-CN-8P and α(S1)-CN-9P were affected by a region on Bos taurus autosome (BTA) 6, but only α(S1)-CN-8P was affected by a region on BTA11 that contains the gene that encodes for β-lactoglobulin (β-LG), and only α(S1)-CN-9P was affected by a region on BTA14 that contains the diacylglycerol acyltransferase 1 (DGAT1) gene. Estimated effects of β-LG protein genotypes showed that only α(S1)-CN-8P was associated with the β-LG A/B polymorphism (g.1772G>A and g.3054C>T); the AA genotype of β-LG was associated with a lower concentration of α(S1)-CN-8P (-0.32% wt/wt) than the BB genotype (+0.41% wt/wt). Estimated effects of DGAT1 K232A genotypes showed that only α(S1)-CN-9P was associated with the DGAT1 gene polymorphism; DGAT1 AA genotype was associated with a higher α(S1)-CN-9P concentration (+0.53% wt/wt) than the DGAT1 KK genotype (-0.44% wt/wt). The results give insight in phosphorylation of α(S1

  6. Solubilization of rehydrated frozen highly concentrated micellar casein for use in liquid food applications.

    Science.gov (United States)

    Lu, Y; McMahon, D J; Metzger, L E; Kommineni, A; Vollmer, A H

    2015-09-01

    Highly concentrated micellar casein concentrate (HC-MCC), a potential ingredient of protein-fortified food, is a gel at cold temperature. It contains ~17 to 21% casein, with most serum proteins and lactose removed by microfiltration and diafiltration, and it is then further concentrated using vacuum evaporation. The HC-MCC can be stored frozen, and our objective was to determine the conditions needed to obtain complete solubility of thawed HC-MCC in water and to understand its gelation upon cooling. Dispersibility (ability to pass through a 250-µm mesh sieve), suspendability (percentage of protein not sedimented at 80 × g within 5min), and solubility (percentage of protein not sedimented at 20,000 × g within 5min) were measured at 4, 12, or 20°C after various mixing conditions. Gelation upon cooling from 50 to 5°C was monitored based on storage (G') and loss (G'') moduli. The gelled HC-MCC was also examined by transmission electron microscopy. Thawed HC-MCC was added to water to reach a protein concentration of 3% and mixed using high shear (7,500rpm) for 1min or low shear (800rpm) for 30min at 4, 12, 20, or 50°C and at pH 6.4 to 7.2. The HC-MCC completely dispersed at 50°C, or at ≤20°C followed by overnight storage at 4°C. Suspendability at 50°C was ~90% whereas mixing at ≤20°C followed by overnight storage resulted in only ~57% suspendability. Solubility followed a similar trend with ~83% at 50°C and only ~29% at ≤20°C. Mixing HC-MCC with 60mM trisodium citrate increased dispersibility to 99% and suspendability and solubility to 81% at 20°C. Cold-gelling temperature, defined as the temperature at which G'=G'' when cooling from 50 to 5°C, was positively correlated with protein level in HC-MCC. Gelation occurred at 38, 28, and 7°C with 23, 20, and 17% of protein, respectively. Gelation was reversible upon heating, although after a second cooling cycle the HC-MCC gel had lower G'. In micrographs of gelled HC-MCC, the casein micelles were

  7. INFLUENCE OF BOVINE AND CAPRINE CASEIN PHOSPHOPEPTIDES DIFFERING IN ALPHA S1-CASEIN CONTENT IN DETERMINING THE ABSORPTION OF CALCIUM FROM BOVINE AND CAPRINE CALCIUM-FORTIFIED MILKS IN RATS

    Science.gov (United States)

    Bovine and caprine milks have a similar overall gross composition, but vary considerably in the ratios of their casein components. These differences cause significant changes in the casein’s ability to bind and stabilize calcium (Ca). It might be expected that these in vitro variations, which are th...

  8. Casein kinases

    DEFF Research Database (Denmark)

    Issinger, O G

    1993-01-01

    subunits are highly conserved during evolution. The relationship between CK-2 alpha from humans and plants is still 73%. Similar relationships are reported for the beta-subunit. Chromosomal assignment of CK-2 alpha shows two gene loci, one of which is a pseudogene. They are located on different chromosomes...... genetic changes are necessarily involved; the observed changes may be entirely due to a signal transduction pathway where CK-2 could be phosphorylated by another kinase(s). CK-2 cDNAs from various organisms have been isolated and characterized. From the deduced amino acid sequence it turns out that CK-2...

  9. Translational and rotational diffusion of flexible PEG and rigid dendrimer probes in sodium caseinate dispersions and acid gels

    NARCIS (Netherlands)

    Salami, S.; Rondeau-Mouro, C.; Barhoum, M.; Duynhoven, van J.P.M.; Mariette, F.

    2014-01-01

    The dynamics of rigid dendrimer and flexible PEG probes in sodium caseinate dispersions and acid gels, including both translational diffusion and rotational diffusion, were studied by NMR. Above the onset of the close-packing limit (C ~ 10 g/100 g H2O), translational diffusion of the probe depended

  10. The Gluten-Free, Casein-Free Diet in Autism: Results of a Preliminary Double Blind Clinical Trial

    Science.gov (United States)

    Elder, Jennifer Harrison; Shankar, Meena; Shuster, Jonathan; Theriaque, Douglas; Burns, Sylvia; Sherrill, Lindsay

    2006-01-01

    This study tested the efficacy of a gluten-free and casein-free (GFCF) diet in treating autism using a randomized, double blind repeated measures crossover design. The sample included 15 children aged 2-16 years with autism spectrum disorder. Data on autistic symptoms and urinary peptide levels were collected in the subjects' homes over the 12…

  11. Removal of High Concentration Chromium by a Foam-separating Technique Using Casein Proteins as a Foaming Reagent

    Science.gov (United States)

    Sugimoto, Futoshi

    Foam separation of high concentration chromium in leather tanning wastewater was investigated using casein protein as a foaming reagent5mL of5w/v% ammonium acetate buffer was added to the sample chromium water. After adjusting the pH to 9.0,4g/L concentrations of casein and gelatin solution were added to recovery the coagulating flocs of chromium resulting foam separation. The sample water containing chromium flocs was incased in reactor, then mixed with distilled water and 1mL of ethanol to sum 200mL total. The foam separation was performed at time intervals of 3min with an air flow rate of 300mL/min. With casein reagent, the removal rate of chromium was not influenced by the presence of NaCl, however, the rate decreased tendency using with the use of gelatin. The proposed method, utilizing 4g/L of casein solution with water, was not influenced by the presence of calcium (<34mM), magnesium (<1mM), carbonate (<0.5mM), bicarbonate (<1.2mM) nor sulfate (<350mM) ions, and is ideal for foam separation in chromium concentrations of about 100mgCr/L.

  12. Dissolution and reconstitution of casein micelle containing dairy powders by high shear using ultrasonic and physical methods.

    Science.gov (United States)

    Chandrapala, Jayani; Martin, Gregory J O; Kentish, Sandra E; Ashokkumar, Muthupandian

    2014-09-01

    The effect of shear on the solubilization of a range of dairy powders was investigated. The rate of solubilization of low solubility milk protein concentrate and micellar casein powders was examined during ultrasonication, high pressure homogenization and high-shear rotor-stator mixing and compared to low-shear overhead stirring. The high shear techniques were able to greatly accelerate the solubilization of these powders by physically breaking apart the powder agglomerates and accelerating the release of individual casein micelles into solution. This was achieved without affecting the structure of the solubilized proteins. The effect of high shear on the re-establishment of the mineral balance between the casein micelles and the serum was examined by monitoring the pH of the reconstituted skim milk powder after prior exposure to ultrasonication. Only minor differences in the re-equilibration of the pH were observed after sonication for up to 3 min, suggesting that the localized high shear forces exerted by sonication did not significantly affect the mass transfer of minerals from within the casein micelles. PMID:24798226

  13. Addendum to "Gluten-Free and Casein-Free Diets in Treatment of Autism Spectrum Disorders: A Systematic Review"

    Science.gov (United States)

    Mulloy, Austin; Lang, Russell; O'Reilly, Mark; Sigafoos, Jeff; Lancioni, Giulio; Rispoli, Mandy

    2011-01-01

    Shortly after the publication of our literature review on gluten-free and casein-free (GFCF) diets in the treatment of autism spectrum disorders (ASD; Mulloy et al., 2010), Whiteley et al. (2010) published the results of experimental evaluation of a GFCF diet. To update our previous literature review, we herein provide an analysis of the Whiteley…

  14. Gluten-Free and Casein-Free Diets in the Treatment of Autism Spectrum Disorders: A Systematic Review

    Science.gov (United States)

    Mulloy, Austin; Lang, Russell; O'Reilly, Mark; Sigafoos, Jeff; Lancioni, Giulio; Rispoli, Mandy

    2010-01-01

    This paper systematically reviews research on the effects of gluten-free and/or casein-free (GFCF) diets in the treatment of ASD. Database, hand, and ancestry searches identified 15 articles for review. Each study was analyzed and summarized in terms of (a) participants, (b) specifics of the intervention, (c) dependent variables, (d) results, and…

  15. Nutritional Impact of a Gluten-Free Casein-Free Diet in Children with Autism Spectrum Disorder

    Science.gov (United States)

    Marí-Bauset, Salvador; Llopis-González, Agustín; Zazpe, Itziar; Marí-Sanchis, Amelia; Suárez-Varela, María Morales

    2016-01-01

    We compared anthropometric values, nutrient intake, the Healthy Eating Index and food variety in children with autism spectrum disorder (ASD), 20 on a gluten-free casein-free (GFCF) diet and 85 on a regular diet in Valencia (Spain) using 3-days food diaries. Those on the GFCF diet had a lower weight, body mass index, and total energy, pantothenic…

  16. Inhibitory effect of fluvoxamine on β-casein expression via a serotonin-independent mechanism in human mammary epithelial cells.

    Science.gov (United States)

    Chiba, Takeshi; Maeda, Tomoji; Kimura, Soichiro; Morimoto, Yasunori; Sanbe, Atsushi; Ueda, Hideo; Kudo, Kenzo

    2015-11-01

    Selective serotonin reuptake inhibitors (SSRIs) are widely used as a first-line therapy in postpartum depression. The objective of this study was to determine the mechanism underlying the inhibitory effects of the SSRI, fluvoxamine, on β-casein expression, an indicator of lactation, in MCF-12A human mammary epithelial cells. Expression levels of serotonin (5-hydroxytryptamine; 5-HT) transporter, an SSRI target protein, and tryptophan hydroxylase 1, a rate-limiting enzyme in 5-HT biosynthesis, were increased in MCF-12A cells by prolactin treatment. Treatment with 1 μM fluvoxamine for 72 h significantly decreased protein levels of β-casein and phosphorylated signal transducer and activator transcription 5 (pSTAT5). Extracellular 5-HT levels were significantly increased after exposure to 1 μM fluvoxamine, in comparison with those of untreated and vehicle-treated cells; however, extracellular 5-HT had little effect on the decrease in β-casein expression. Expression of glucose-related protein 78/binding immunoglobulin protein, a regulator of endoplasmic reticulum (ER) stress, was significantly increased after treatment with 1 μM fluvoxamine for 48 h. Exposure to tunicamycin, an inducer of ER stress, also decreased expression of β-casein and pSTAT5 in a manner similar to fluvoxamine. Our results indicate that fluvoxamine suppresses β-casein expression in MCF-12A cells via inhibition of STAT5 phosphorylation caused by induction of ER stress. Further studies are required to confirm the effect of fluvoxamine on the function of mammary epithelial cells. PMID:26415980

  17. Casein Films: The Effects of Formulation, Environmental Conditions and the Addition of Citric Pectin on the Structure and Mechanical Properties

    Directory of Open Access Journals (Sweden)

    Laetitia M. Bonnaillie

    2014-07-01

    Full Text Available Thin casein films for food packaging applications reportedly possess good strength and low oxygen permeability, but low elasticity and high sensitivity to moisture. Modifying the films to target specific behaviors depending on environmental conditions can enable a variety of commercial applications for casein-based films. The mechanical properties of solvent-cast (15% solids calcium-caseinate/glycerol films (CaCas:Gly ratio of 3:1 were characterized as a function of processing and environmental conditions, including film thickness, solution formulation and ambient humidity (from 22% to 70% relative humidity (RH at ~20 °C. At constant RH, the elongation at break (EAB had a strong positive dependence on the film thickness. When RH increased, the tensile strength (TS and modulus (E decreased approximately linearly, while EAB increased. From 0.05% to 1% (w/w of citric pectin (CP was then incorporated into CaCas/Gly films following seven different formulations (mixing sequences, to alter the protein network and to evaluate the effects of CP on the tensile properties of CaCas/Gly/CP films. At constant film thickness and ~60% RH, the addition of 0.1% or 1.0% CP to the films considerably increased or decreased EAB, TS and E in different directions and to different extents, depending on the formulation, while optical micrographs also showed vastly differing network configurations, suggesting complex formulation- and stoichiometry-dependent casein-pectin interactions within the dried films. Depending on the desired film properties and utilization conditions, pectin may be a useful addition to casein film formulations for food packaging applications.

  18. Changes of pH in β-Lactoglobulin and β-Casein Solutions during High Pressure Treatment

    Directory of Open Access Journals (Sweden)

    Karsten Olsen

    2015-01-01

    Full Text Available The pH changes in the milk systems, β-lactoglobulin B, β-casein, and mixture of β-lactoglobulin and β-casein (pH 7 and ionic strength 0.08 M were measured in situ during increasing pressure up to 500 MPa. An initial decrease to pH 6.7 was observed from 0.1 to 150 MPa for β-lactoglobulin, followed by an increase to pH 7.3 at 500 MPa. The initial decrease is suggested to be caused by the deprotonation of histidine, while the increase is suggested to result from an increase of hydroxide ions due to the loss of tertiary structure. The change in pH of the β-casein solution displayed an almost linear increasing pressure dependency up to a pH of 7.7 at 500 MPa. The limited tertiary structure of β-casein could allow exposure of all amino acids; thus the increase of pH can be caused by binding of water protons resulting in an increase of hydroxide ions. Addition of β-casein to β-lactoglobulin (1:1 was found to suppress the initial pH decrease found for the β-lactoglobulin solution. The pH change of the mixture did not suggest any intermolecular interaction, and a simple additive model is proposed to calculate the pH change of the mixture from the corresponding individual samples.

  19. Casein and soy protein meals differentially affect whole-body and splanchnic protein metabolism in healthy humans.

    Science.gov (United States)

    Luiking, Yvette C; Deutz, Nicolaas E P; Jäkel, Martin; Soeters, Peter B

    2005-05-01

    Dietary protein quality is considered to be dependent on the degree and velocity with which protein is digested, absorbed as amino acids, and retained in the gut as newly synthesized protein. Metabolic animal studies suggest that the quality of soy protein is inferior to that of casein protein, but confirmatory studies in humans are lacking. The study objective was to assess the quality of casein and soy protein by comparing their metabolic effects in healthy human subjects. Whole-body protein kinetics, splanchnic leucine extraction, and urea production rates were measured in the postabsorptive state and during 8-h enteral intakes of isonitrogenous [0.42 g protein/(kg body weight . 8 h)] protein-based test meals, which contained either casein (CAPM; n = 12) or soy protein (SOPM; n = 10) in 2 separate groups. Stable isotope techniques were used to study metabolic effects. With enteral food intake, protein metabolism changed from net protein breakdown to net protein synthesis. Net protein synthesis was greater in the CAPM group than in the SOPM group [52 +/- 14 and 17 +/- 14 nmol/(kg fat-free mass (FFM) . min), respectively; P CAPM (P = 0.07). Absolute splanchnic extraction of leucine was higher in the subjects that consumed CAPM [306 +/- 31 nmol/(kg FFM . min)] vs. those that consumed SOPM [235 +/- 29 nmol/(kg FFM . min); P < 0.01]. In conclusion, a significantly larger portion of soy protein is degraded to urea, whereas casein protein likely contributes to splanchnic utilization (probably protein synthesis) to a greater extent. The biological value of soy protein must be considered inferior to that of casein protein in humans. PMID:15867285

  20. Viability of the microencapsulation of a casein hydrolysate in lipid microparticles of cupuacu butter and stearic acid

    Directory of Open Access Journals (Sweden)

    Samantha Cristina Pinho

    2013-04-01

    Full Text Available Normal 0 21 false false false PT-BR X-NONE X-NONE Solid lipid microparticles produced with a mixture of cupuacu butter and stearic acid were used to microencapsulate a commercial casein hydrolysate (Hyprol 8052. The composition of the lipid matrix used for the production of the lipid microparticles was chosen according to data on the wide angle X-ray diffraction (WAXD and differential scanning calorimetry (DSC of bulk lipid mixtures, which indicated that the presence of 10 % cupuacu butter was sufficient to significantly change the crystalline arrangement of pure stearic acid. Preliminary tests indicated that a minimum proportion of 4 % of surfactant (polysorbate 80 was necessary to produce empty spherical lipid particles with average diameters below 10 mm. The lipid microparticles were produced using 20 % cupuacu butter and 80 % stearic acid and then stabilized with 4 % of polysorbate 80, exhibiting an encapsulation efficiency of approximately 74 % of the casein hydrolysate. The melting temperature of the casein hydrolysate-loaded lipid microparticles was detected at 65.2 °C, demonstrating that the particles were solid at room temperature as expected and indicating that the incorporation of peptides had not affected their thermal behavior. After 25 days of storage, however, there was a release of approximately 30 % of the initial amount of encapsulated casein hydrolysate. This release was not thought to have been caused by the liberation of encapsulated casein hydrolysate. Instead, it was attributed to the possible desorption of the adsorbed peptides present on the surface of the lipid microparticles.

  1. Isolation and characterization of human cDNA clones encoding the α and the α' subunits of casein kinase II

    International Nuclear Information System (INIS)

    Casein kinase II is a widely distributed protein serine/threonine kinase. The holoenzyme appears to be a tetramer, containing two α or α' subunits (or one of each) and two β subunits. Complementary DNA clones encoding the subunits of casein kinase II were isolated from a human T-cell λgt 10 library using cDNA clones isolated from Drosophila melanogasten. One of the human cDNA clones (hT4.1) was 2.2 kb long, including a coding region of 1176 bp preceded by 156 bp (5' untranslated region) and followed by 871 bp (3' untranslated region). The hT4.1 close was nearly identical in size and sequence with a cDNA clone from HepG2 human hepatoma cultured cells. Another of the human T-cell cDNA clones (hT9.1) was 1.8 kb long, containing a coding region of 1053 bp preceded by 171 by (5' untranslated region) and followed by 550 bp (3' untranslated region). Amino acid sequences deduced from these two cDNA clones were about 85% identical. Most of the difference between the two encoded polypeptides was in the carboxy-terminal region, but heterogeneity was distributed throughout the molecules. Partial amino acid sequence was determined in a mixture of α and α' subunits from bovine lung casein kinase II. The bovine sequences aligned with the 2 human cDNA-encoded polypeptides with only 2 discrepancies out of 535 amino acid positions. This confirmed that the two human T-cell cDNA clones encoded the α and α' subunits of casein kinase II. These studies show that there are two distinct catalytic subunits for casein II (α and α') and that the sequence of these subunits is largely conserved between the bovine and the human

  2. Enhanced removal of detergent and recovery of enzymatic activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis: UUse of casein in gel wash buffer

    International Nuclear Information System (INIS)

    The inclusion of 1% casein or bovine serum albumin in buffer used to reactivate enzymes subjected to sodium dodecyl sulfate (SDS)-polyacrylamide electrophoresis resulted in accelerated removal of SDS and restoration of nuclease and beta-galactosidase enzyme activities. Nuclease and beta-galactosidase activities which are absent from gels after longer wash procedures are detectable with this technique. Enzyme activity in gels prepared with SDS which contained inhibitory contaminants was partially restored by the casein wash procedure. The threshold of detection of two-dimensionally separated deoxyribonuclease I using the casein wash procedure was 1 picogram

  3. Extreme frequencies of the αs1-casein "null" variant in milk from Norwegian dairy goats-- implications for milk composition, micellar size and renneting properties

    OpenAIRE

    Devold, Tove G.; Nordbø, Ragnhild; Langsrud, Thor; Svenning, Cecilie; Jansen Brovold, Margreet; Sørensen, Esben,; Christensen, Brian; Ådnøy, Tormod; Vegarud, Gerd

    2011-01-01

    Caprine alpha(s1)-casein polymorphism is one of the key factors which determines important technological properties of milk, such as rennetability and cheese yield. The indigenous Norwegian goat breed is characterized by a remarkably high frequency (> 70%) of a "null" variant of alpha(s1)-casein unique to this population. This mutation, referred to as the exon 12 D allele, involves a single base deletion in exon 12 of the alpha(s1)-casein gene. The aim of this study was to provide more detail...

  4. Nuclear magnetic resonance study of the conformation and dynamics of beta-casein at the oil/water interface in emulsions.

    OpenAIRE

    ter Beek, L C; Ketelaars, M; McCain, D C; Smulders, P E; Walstra, P.; Hemminga, M.A.

    1996-01-01

    A (13)C and (31)P nuclear magnetic resonance (NMR) study has been carried out on beta-casein adsorbed at the interface of a tetradecane/water emulsion. (13)C NMR spectra show signals from the carbonyl, carboxyl, aromatic, and C alpha carbons in beta-casein, well resolved from solvent resonances. Only a small fraction of all carbon atoms in beta-casein contribute to detectable signals; intensity measurements show that the observable spectrum is derived from about 30 to 40 amino acid residues.(...

  5. A Review of Casein Phosphopeptide- Amorphous Calcium Phosphate (CPP-ACP) and Enamel Remineralization.

    Science.gov (United States)

    Gupta, Neetu; Mohan Marya, Charu; Nagpal, Ruchi; Singh Oberoi, Sukhvinder; Dhingra, Chandan

    2016-01-01

    White-spot lesions are the earliest macroscopic evidence of enamel caries. In such a situation, the enamel surface layer stays intact during subsurface demineralization, but without treatment the subsurface loss will continue, and eventually the surface layer will collapse and lead to a cavity formation. By introducing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) at this stage the lesion can be reversed. CPP-ACP is a unique, naturally derived protein-based remineralizing technology that is now used globally in chewing gums and topical creams. The aim of this review is to expound on the potential for reversal of demineralization through the use of products such as CPP-ACP, and to provide guidance to clinicians considering remineralization as a viable treatment option. PMID:26863219

  6. Hematein, a casein kinase II inhibitor, inhibits lung cancer tumor growth in a murine xenograft model.

    Science.gov (United States)

    Hung, Ming-Szu; Xu, Zhidong; Chen, Yu; Smith, Emmanuel; Mao, Jian-Hua; Hsieh, David; Lin, Yu-Ching; Yang, Cheng-Ta; Jablons, David M; You, Liang

    2013-11-01

    Casein kinase II (CK2) inhibitors suppress cancer cell growth. In this study, we examined the inhibitory effects of a novel CK2 inhibitor, hematein, on tumor growth in a murine xenograft model. We found that in lung cancer cells, hematein inhibited cancer cell growth, Akt/PKB Ser129 phosphorylation, the Wnt/TCF pathway and increased apoptosis. In a murine xenograft model of lung cancer, hematein inhibited tumor growth without significant toxicity to the mice tested. Molecular docking showed that hematein binds to CK2α in durable binding sites. Collectively, our results suggest that hematein is an allosteric inhibitor of protein kinase CK2 and has antitumor activity to lung cancer. PMID:24008396

  7. Drying Technology of Angiotensin Converting Enzyme Inhibitory Peptide Derived from Bovine Casein

    Institute of Scientific and Technical Information of China (English)

    JIANG Zhanmei; Hue Guicheng; TIAN Be

    2009-01-01

    Drying technology of angiotensin converting enzyme (ACE) inhibitory peptides derived from bovine casein was investigated. No significance was observed on ACE inhibitory activity of products prepared by spay drying and freeze drying (P>0.05). Spay drying was the best drying process for practical industry production. The inlet temperature ranged from 140℃ to 160℃ and the exit temperature ranged from 70℃ to 90℃ during the spay drying process. Under the optimal eonditious, scale-up of angiotensin converted enzyme inhibitory peptide from 1 L to 10 L and the experiment was successively conducted. Peptide yield was 29% and half inhibitory concentration(IC50) was 0.53g·L-1.

  8. Quantitative analysis of complex casein hydrolysates based on chromatography and membrane

    Institute of Scientific and Technical Information of China (English)

    Qi Wei; Yu Yanjun; He Zhimin

    2006-01-01

    The enzymatic hydrolysates of casein are so complex that there is no effective method to do quantitative analysis.The common techniques,such as high performance chromatography and SDS-PAGE,can only carry out qualitative analysis.On the basis of membrane separation and high performance size exclusion chromatography (HPSEC),standard peptides with different molecular mass range were prepared,and the linear relationships between mass concentration of the standard peptides and the ultraviolet absorption of corresponding peak areas were established.Consequently,mass concentration of the different hydrolysates at different reaction times could be accurately calculated.The combination of chromatography and membrane separation is of great importance to the quantitative analysis of the complex hydrolysates,which can also be applied to the other macromolecular systems,such as carbohydrates.

  9. The increasing of enamel calcium level after casein phosphopeptideamorphous calcium phosphate covering

    Directory of Open Access Journals (Sweden)

    Widyasri Prananingrum

    2012-06-01

    Full Text Available Background: Caries process is characterized by the presence of demineralization. Demineralization is caused by organic acids as a result of carbohydrate substrate fermentation. Remineralization is a natural repair process for non-cavitated lesions. Remineralization occurs if there are Ca2+ and PO43- ions in sufficient quantities. Casein-amorphous calcium phosphate phosphopeptide (CPP-ACP is a paste material containing milk protein (casein, that actually contains minerals, such as calcium and phosphate. The casein ability to stabilize calcium phosphate and enhance mineral solubility and bioavailability confers upon CPP potential to be biological delivery vehicles for calcium and phosphate. Purpose: The aim of this study was to determine the calcium levels in tooth enamel after being covered with CPP-ACP 2 times a day for 3, 14 and 28 days. Methods: Sample were bovine incisors of 3 year old cows divided into 4 groups, namely group I as control group, group II, III and IV as treatment groups covered with CPP-ACP 2 times a day. All of those teeth were then immersed in artificial saliva. Group II was immersed for 3 days, while group III was immersed for 14 days, and group IV was immersed for 28 days. One drop of CPP-ACP was used to cover the entire labial surface of teeth. The measurement of the calcium levels was then conducted by using titration method. All data were analyzed by One- Way ANOVA test with 5% degree of confidence. Results: The results showed significant difference of the calcium levels in tooth enamel of those groups after covered with CPP-ACP 2 times a day for 3, 14 and 28 days (p = 0.001. There is also significant difference of the calcium levels in tooth enamel of those treatment groups and the control group (p = 0.001. Conclusion: The calcium levels of tooth enamel are increased after covered with CPP-ACP 2 times a day for 3, 14 and 28 days.Latar belakang: Proses terjadinya karies gigi ditandai oleh adanya demineralisasi

  10. Treatment of post-orthodontic white spot lesions with casein phosphopeptide-stabilised amorphous calcium phosphate

    DEFF Research Database (Denmark)

    Bröchner, Ann; Christensen, Carsten; Kristensen, Bjarne;

    2010-01-01

    This study aims to investigate the effect of topical applications of 10% casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSL) detected after treatment with fixed orthodontic appliances. Sixty healthy adolescents with >/=1 clinically visible WSL at debonding were...... recruited and randomly allocated to a randomised controlled trial with two parallel groups. The intervention group was instructed to topically apply a CPP-ACP -containing agent (Tooth Mousse, GC Europe) once daily and the subjects of the control group brushed their teeth with standard fluoride toothpaste...... included in the final evaluation. A statistically significant (p <0.05) regression of the WSL was disclosed in both study groups compared to baseline, but there was no difference between the groups. The mean area of the lesions decreased by 58% in the CPP-ACP group and 26% in the fluoride group (p = 0...

  11. Circadian and pharmacological regulation of casein kinase I in the hamster suprachiasmatic nucleus

    Indian Academy of Sciences (India)

    Patricia V. Agostino; Santiago A. Plano; Diego A. Golombek

    2008-12-01

    In mammals, the mechanism for the generation of circadian rhythms and entrainment by light–dark (LD) cycles resides in the hypothalamic suprachiasmatic nuclei (SCN), and the principal signal that adjusts this biological clock with environmental timing is the light:dark cycle. Within the SCN, rhythms are generated by a complex of molecular feedback loops that regulate the transcription of clock genes, including per and cry. Posttranslational modification plays an essential role in the regulation of biological rhythms; in particular, clock gene phosphorylation by casein kinase I, both epsilon (CKI) and delta (CKI), regulates key molecular mechanisms in the circadian clock. In this paper, we report for the first time that CKI activity undergoes a significant circadian rhythm in the SCN (peaking at circadian time 12, the start of the subjective night), and its pharmacological inhibition alters photic entrainment of the clock, indicating that CKI may be a key element in this pathway.

  12. Genetic polymorphism of the K-casein (CSN3 gene in goats reared in Southern Italy

    Directory of Open Access Journals (Sweden)

    Fabio Pilla

    2010-01-01

    Full Text Available K-casein (K-CN represents one of the most important proteins determining the manufacturing properties of milk,because of its essential role in micelle formation and stabilisation.Several genetic variants of K-CN have been described in goats. To investigate the occurrence of seven alleles and theirdistribution among breeds, a total of 170 animals, from six different breeds reared in Italy (Cilentana Nera, Derivata diSiria, Maltese, Jonica, Garganica and Cashmere, have been analysed in this paper by the primer extension method.Alleles A and B were found to be the most represented in all the analysed breeds; allele D is present only in Maltese andCashmere animals with a very low frequency; while allele G has been found in all but two (Garganica and Cashmerebreeds. Alleles C, E and F were not present in the material used for this study.

  13. Casein kinase II as a potentially important enzyme concerned with signal transduction

    International Nuclear Information System (INIS)

    At least two of the proteins that undergo changes in serine (threonine) phosphorylation as a result of insulin action are substrates for the protein serine kinase known as casein kinase II (CK-II). Although CK-II has been studied for a number of years, its physiological functions are not well understood. Several developments, which have stimulated a renewed interest in this particular protein kinase, are reviewed in this paper. Topics discussed include a review of the properties of CK-II, the regulation of CK-II levels, a computer search for potential substrates of CK-II, and phosphorylation of nuclear oncoproteins by CK-II. Its unique features described in the paper make CK-II a candidate in controlling aspects of gene transcription

  14. Isolation and characterization of recombinant human casein kinase II subunits alpha and beta from bacteria

    DEFF Research Database (Denmark)

    Grankowski, N; Boldyreff, B; Issinger, O G

    1991-01-01

    cDNA encoding the casein kinase II (CKII) subunits alpha and beta of human origin were expressed in Escherichia coli using expression vector pT7-7. Significant expression was obtained with E. coli BL21(DE3). The CKII subunits accounted for approximately 30% of the bacterial protein; however, most...... of the expressed proteins were produced in an insoluble form. The recombinant CKII alpha subunit was purified by DEAE-cellulose chromatography, followed by phosphocellulose and heparin-agarose chromatography. The recombinant CKII beta subunit was extracted from the insoluble pellet and purified in a...... single step on phosphocellulose. From 10 g bacterial cells, the yield of soluble protein was 12 mg alpha subunit and 5 mg beta subunit. SDS/PAGE analysis of the purified recombinant proteins indicated molecular masses of 42 kDa and 26 kDa for the alpha and beta subunits, respectively, in agreement with...

  15. IDENTIFICATION OF BETA-LACTOGLOBULIN AND KAPPA-CASEIN GENOTYPES IN CATTLE

    Directory of Open Access Journals (Sweden)

    TESIO C.D.

    2007-01-01

    Full Text Available Beta-lactoglobulin (b-Lg and kappa-casein (k-Cn are two of the most importantproteins in the mammals’ milk synthesized by the epithelial cells of the mammaryglands. They play a crucial role in the milk quality and coagulation process(production of cheese and butter. The PCR-RFLP test was performed to distinguishthe different alleles in a population of Romanian Black Spotted cattle, a dairy breed.Genetic polymorphism was detected by digestion with the endonucleases Hae III (b-Lg and Hinf I (k-Cn, followed by electrophoresis in agarose high resolution gelstained with ethidium bromide. Fifty DNA samples from Romanian Black Spottedbreed were analyzed for A and B variants. This simple PCR-RFLP test makesfeasible the inclusion of b-Lg and k-Cn genotypes in breeding plans and cattleselection.

  16. Formulation and characterization of nanoencapsulated curcumin using sodium caseinate and its incorporation in ice cream.

    Science.gov (United States)

    Kumar, Deep Diyuti; Mann, Bimlesh; Pothuraju, Ramesh; Sharma, Rajan; Bajaj, Rajesh; Minaxi

    2016-01-01

    In the present investigation, the preparation and characterization of a curcumin nanoemulsion with milk protein (sodium caseinate) and its incorporation into ice cream were undertaken. Among the different combinations, the most stable formulation was observed using milk fat (8%), medium chain triglycerides (2%), curcumin (0.24%) and sodium caseinate (6%) with a mean particle size of 333.8 ± 7.18 nm, a zeta potential of -44.1 ± 0.72 mV and an encapsulation efficiency of 96.9 ± 0.28%. The effect of different processing conditions (heating, pH and ionic strength) on the particle size distribution and zeta potential of the nanoemulsion was evaluated. During heat treatment, the particle size of the nanoemulsion was increased from 333.8 ± 7.18 to 351.1 ± 4.04 nm. The nanoemulsion was destabilized at pH 4.6 and the particle size increased above and below pH 5.0. However, there was a slight increase in the particle size with a change in the ionic concentration. The release kinetics data suggested that in simulated gastro-intestinal digestion, the nanoemulsion was stable against pepsin digestion (a 5.25% release of curcumin), while pancreatic action led to a 16.12% release of curcumin from the nanoemulsion. Finally, our formulation was successfully incorporated into ice cream and the sensory attributes were evaluated. No significant difference was observed in the scores of the sensory attributes between the control and ice cream prepared with a curcumin nanoemulsion. Moreover, the encapsulation efficiency of the curcumin incorporated into the ice cream was 93.7%, which indicates that it can withstand the processing conditions. The findings suggest that ice cream is a suitable dairy product for the delivery of lipophilic bioactive components (curcumin) which can be used for therapeutic purposes. PMID:26501983

  17. Structure and gelation properties of casein micelles doped with curcumin under acidic conditions.

    Science.gov (United States)

    Khanji, Aya N; Michaux, Florentin; Jasniewski, Jordane; Petit, Jeremy; Lahimer, Emna; Cherif, Mohamed; Salameh, Dominique; Rizk, Toufic; Banon, Sylvie

    2015-12-01

    In this study, the ability of micellar casein (MC) to interact with curcumin during acidification and to produce acid gel was investigated. Steady-state fluorescence spectroscopy of curcumin variation and fluorescence quenching of caseins upon binding with curcumin molecules were evidenced. Increasing the temperature from 20 to 35 °C enhanced MC-curcumin interactions as reflected by the increase in the binding constant from 0.6 ± 0.3 × 10(4) to 6.6 ± 0.6 × 10(4) M(-1). From changes in entropy, enthalpy and Gibbs free energy, hydrophobic interactions were proposed as major binding forces. Static fluorescence MC quenching was demonstrated for the MC-curcumin complex during acidification. From pH 7.4 to pH 5.0, the binding site numbers varied in the range from 1.25 ± 0.05 to 1.49 ± 0.05 and the binding constant kb varied from 3.9 ± 0.4 × 10(4) to 7.5 ± 0.7 × 10(4) M(-1). Small angle X-ray scattering profiles demonstrated that the MC internal structure was unchanged upon curcumin binding. The ζ-potential value of curcumin-doped MC indicated that curcumin did not modify the global charge of MC particles. Acid gelation studied by oscillation rheology and static multiple light scattering at 20 and 35 °C led to a similar behavior for native and curcumin-doped MC suspensions. For the first time, it was demonstrated that the colloidal and functional properties of MC were unchanged when doped with curcumin during acidification. PMID:26419313

  18. Study on genetic variability at αS1 and αS2 casein loci in autochthonous goat populations of the Lombardy Alps

    Directory of Open Access Journals (Sweden)

    P. Crepaldi

    2011-03-01

    Full Text Available In cattle, sheep and goats (Hayes et al., 1993 the casein genes form a cluster, on the chromosome 6, of four tightly linked genes, whose sequence is: αS1-β-αS2-k (Threadgill and Womack, 1990; Ferretti et al., 1990; Gallagher et al., 1994; Rijnkles et al., 1997. For this reason the alleles at these loci are generally inherited all together (Di Stasio and Mariani, 2000, with low recombination frequency. Thus it is worth noting the importance of describing the casein haplotypes namely the inherited combination of closely linked alleles (King and Stansfield, 1985; Meuwissen and Goddard, 2001. The knowledge of the casein allele associations could be important for the selection. In goat milk the casein alleles are related to important differences in the level of synthesis of the respective gene products and in milk chemical-physical properties........

  19. A novel method for in Situ detection of hydrolyzable casein fragments in a cheese matrix by antibody phage display technique and CLSM

    DEFF Research Database (Denmark)

    Duan, Zhi; Brüggemann, Dagmar Adeline; Siegumfeldt, Henrik

    2009-01-01

    A novel method to monitor in situ hydrolyzable casein fragments during cheese ripening by using immunofluorescent labeling and confocal laser scanning microscopy (CLSM) was developed. Monoclonal single chain variable fragments of antibody (scFvs) were generated by antibody phage display toward...... three small synthetic peptides of the alpha(s1)-casein sequence. These peptides traverse enzymatic cleavage sites of casein during cheese ripening. The specificity of the generated anti-peptide antibodies was determined by ELISA and Western blot. Finally, an immunofluorescent labeling protocol was...... successfully developed for the detection of scFvs binding to different alpha(s1)-casein fragments inside a cheese matrix by CLSM. To our knowledge, this is the first demonstrated immunofluorescent labeling method for in situ analysis of proteolysis phenomena in the cheese matrix. Additionally, this technique...

  20. Effect Of Five Proteases Including Alcalase, Flavourzyme, Papain, Proteinase K And Trypsin On Antioxidative Activities Of Casein Hydrolysate From Goat Milk

    OpenAIRE

    Shu Guowei; Zhang Qian; Chen He; Wan Hongchang; Li Hong

    2015-01-01

    Oxidation was related to the pathogenesis of human diseases. Adequate intake of antioxidant activity of food can reduce the levels of free radicals, prevent lipid peroxidation, and help the body against diseases. In the paper, casein from goat milk was hydrolyzed by five commercial proteases, namely, Alcalase, flavourzyme, papain, proteinase K and trypsin. The antioxidant activities of casein hydrolysates were assessed by evaluating hydrolysis degree, DPPH radical-scavenging activity, metal-c...

  1. Quantification of bovine β-casein allergen in baked foodstuffs based on ultra-performance liquid chromatography with tandem mass spectrometry

    OpenAIRE

    Chen, Qi; Zhang, Jingshun; Ke, Xing; Lai, Shiyun; Tao, Baohua; Yang, Jinchuan; Mo, Weimin; Ren, Yiping

    2014-01-01

    The quantification of allergens in food including baked food matrices is of great interest. The aim of the present study was to describe a non-immunologic method to quantify bovine β-casein using ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) in multiple reaction monitoring (MRM) mode. Eight of 10 theoretical peptides from β-casein after tryptic digestion were compared and MRM methods were developed to determine five signature peptides. The ...

  2. Influence of human diets containing casein, soy protein isolate, and soy protein concentrate on serum cholesterol and lipoproteins in humans, rabbits and rats

    OpenAIRE

    Raaij, van, J.M.A.

    1982-01-01

    It is well known that feeding animals such as rabbits with semipurified diets containing animal proteins, as for example casein, results in hypercholesterolemia and atherosclerosis. On the other hand, diets containing vegetable proteins such as soybean protein maintain low levels of serum cholesterol. Little is known about the effects of the type of protein in the diet in humans.This thesis deals with the effects of casein and soy protein on serum cholesterol and lipoproteins, as observed in ...

  3. Proving the adulteration of ewe and goat cheeses with cow milk using the reference method of isoelectric focusing of γ-casein

    Directory of Open Access Journals (Sweden)

    Nataša Mikulec

    2013-08-01

    Full Text Available The aim of this study was to introduce a reference method for the detection of cow milk in ewe and goat cheeses (EC 273/08 in order to protect manufacturers and consumers from adulterations and imitations, and to ensure alignment with the demands of domestic and EU markets. The method includes isolation of casein from cheese, isoelectric focusing of γ2- and γ3-casein originating from the hydrolysis of β-casein by plasmin, the detection and quantitative determination of γ-casein in cow, ewe and goat cheese by densitometry. Ewe or goat cheese products with a minimum of 1 % of cow milk are considered to be adulterated. For the quantitative determination of cow, ewe and goat milk in cheeses, standard mixtures of cow-ewe and cow-goat milk were made by adding 0; 0.5; 1; 2; 5; 10; 25; 50; 75 and 100 % (v/v of cow milk. The quantification was performed by determining the peak area ratio of cow γ-casein in comparison to ewe/goat casein in prepared standard cheeses. The calibration curves were calculated based on the relation of the peak area ratio of cow γ-caseins (calculated as the percentage of total γ-caseins in contrast to the relative content (% of cow milk in the mixture. The method proved to be adequate for the detection of raw and heat-treated cow milk in fresh and ripened cheeses made from ewe or goat milk, or a mixture of ewe and goat milk.

  4. AlphaS1-casein deficiency recorded for defective genotypes induces a chronic ER stress and deep changes in milk composition, signing a singular secretion process in goats

    OpenAIRE

    Martin, Patrice; B. Badaoui; Barile, Daniela; Lahouassa, Hichem; Beauvallet, Christian; Cebo, Christelle; Leroux, Christine; Chanat, Eric

    2013-01-01

    The extensive polymorphism recorded at the CSN1S1 locus has been shown to influence goat milk composition and its technological properties. A deficit in alphaS1-casein is responsible for the accumulation of immature caseins in distended rough endoplasmic reticulum (ER) cisternae of mammary epithelial cells (MEC). This triggers a chronic ER stress which induces, in turn, an adaptive unfolded protein response (UPR). Our data strongly suggest: i) the existence of a membraneassociated form of alp...

  5. Prolactin and glucocorticoid hormones synergistically induce expression of transfected rat beta-casein gene promoter constructs in a mammary epithelial cell line.

    OpenAIRE

    Doppler, W; Groner, B; Ball, R K

    1989-01-01

    We have detected hormone response elements in the promoter region of the rat beta-casein gene that confer the synergistic action of prolactin and glucocorticoid hormones upon transcription of chimeric gene constructs. A 2800-base-pair (bp) rat beta-casein gene fragment containing 2300 bp of 5' flanking sequence was placed in front of a chloramphenicol acetyltransferase (CAT) reporter gene and stably transfected into the mouse mammary epithelial cell line HC11. Addition of prolactin or dexamet...

  6. Stimulation of casein kinase II by epidermal growth factor: relationship between the physiological activity of the kinase and the phosphorylation state of its beta subunit.

    OpenAIRE

    Ackerman, P; Glover, C V; Osheroff, N

    1990-01-01

    To determine relationships between the hormonal activation of casein kinase II and its phosphorylation state, epidermal growth factor (EGF)-treated and EGF-naive human A-431 carcinoma cells were cultured in the presence of [32P]orthophosphate. Immunoprecipitation experiments indicated that casein kinase II in the cytosol of EGF-treated cells contained approximately 3-fold more incorporated [32P]phosphate than did its counterpart in untreated cells. Levels of kinase phosphorylation paralleled ...

  7. Mesenteric IL-10-producing CD5+ regulatory B cells suppress cow’s milk casein-induced allergic responses in mice

    Science.gov (United States)

    Kim, A-Ram; Kim, Hyuk Soon; Kim, Do Kyun; Nam, Seung Taek; Kim, Hyun Woo; Park, Young Hwan; Lee, Dajeong; Lee, Min Bum; Lee, Jun Ho; Kim, Bokyung; Beaven, Michael A.; Kim, Hyung Sik; Kim, Young Mi; Choi, Wahn Soo

    2016-01-01

    Food allergy is a hypersensitive immune reaction to food proteins. We have previously demonstrated the presence of IL-10-producing CD5+ B cells and suggested their potential role in regulating cow’s milk casein allergy in humans and IgE-mediated anaphylaxis in mice. In this study, we determined whether IL-10-producing CD5+ regulatory B cells control casein-induced food allergic responses in mice and, if so, the underlying mechanisms. The induction of oral tolerance (OT) by casein suppressed casein-induced allergic responses including the decrease of body temperature, symptom score, diarrhea, recruitment of mast cells and eosinophils into jejunum, and other biological parameters in mice. Notably, the population of IL-10-producing CD5+ B cells was increased in mesenteric lymph node (MLN), but not in spleen or peritoneal cavity (PeC) in OT mice. The adoptive transfer of CD5+ B cells from MLN, but not those from spleen and PeC, suppressed the casein-induced allergic responses in an allergen-specific and IL-10-dependent manner. The inhibitory effect of IL-10-producing CD5+ B cells on casein-induced allergic response was dependent on Foxp3+ regulatory T cells. Taken together, mesenteric IL-10-producing regulatory B cells control food allergy via Foxp3+ regulatory T cells and could potentially act as a therapeutic regulator for food allergy. PMID:26785945

  8. Study on Casein Micelles in Raw and Pasteurized Milk in Native State and During the First Stage of the Enzymatic Coagulation by SEM and TEM

    Directory of Open Access Journals (Sweden)

    H. Lamea

    2005-10-01

    Full Text Available In this study changes in microstructure of casein micelles in raw and pasteurized milk were investigated by SEM and TEM. Milk was pasteurized by L.T.L.T. method (Temperature: 63 oC_ Time:30 minute.Samples of raw and pasteurized milk were taken from research pilot plant of The College of Agriculture. Each sample was divided into two parts. One part of each sample was directly prepared for SEM and TEM, whereas the second part were renneted at 2 oC and kept at this temperature for 24 h. and then prepared for evaluation by electron microscopes. Results indicated that in native state, particularly in raw milk casein micelles are in spherical shape with smooth surface and in the single form. The samples, which stored in the cold were under the effect of the rennet, encounter of filamentous status that might be due to the changes occurred in kappa-casein following the action of rennet along with partial removal of beta-casein and micellar calcium phosphate. Micrographs of casein micelles in pasteurized milk indicated that thermal condition during pasteurization had influenced on increasing of casein micelles size.

  9. Effects of sodium caseinate concentration and storage conditions on the oxidative stability of oil-in-water emulsions.

    Science.gov (United States)

    O' Dwyer, Sandra P; O' Beirne, David; Eidhin, Deirdre Ní; O' Kennedy, Brendan T

    2013-06-01

    The oxidative stability of various oils (sunflower, camelina and fish) and 20% oil-in-water (O/W) emulsions, were examined. The mean particle size decreased from 1179 to 325 nm as sodium caseinate (emulsifier) concentration was increased from 0.25% to 3% in O/W emulsions (P<0.05). Increasing the microfluidisation pressure from 21 to 138 MPa, resulted in a particle size decrease from 289 to 194 nm (P<0.05). Emulsified oils had lower detectable lipid hydroperoxide and p-Anisidine values than their corresponding bulk oils (P<0.05). The lipid hydroperoxide and p-Anisidine values of emulsions generally decreased as sodium caseinate concentration increased, and similarly decreased as microfluidisation pressure increased (P<0.05). Increasing storage temperature of the emulsions from 5 to 60°C, resulted in lower detectable lipid oxidation products during storage (P<0.05). PMID:23411225

  10. Characterization of the alpha and beta subunits of casein kinase 2 by far-UV CD spectroscopy

    DEFF Research Database (Denmark)

    Issinger, O G; Brockel, C; Boldyreff, B; Pelton, J T

    1992-01-01

    Although Chou-Fasman calculations of the secondary structure of recombinant casein kinase 2 subunits alpha and beta suggest they have a similar overall conformation, circular dichroism (CD) studies show that substantial differences in the conformation of the two subunits exist. In addition......, no changes in the far-UV CD spectrum of the alpha subunit are observed in the presence of casein or the synthetic decapeptide substrate RRRDDDSDDD. Furthermore, the alpha-helical structure of the alpha subunit (but not the beta subunit) can be increased in the presence of stoichiometric amounts of...... heparin, presumably by its binding to the polylysine stretch at amino acid positions 74-77. Heat denaturation experiments (25-90 degrees C) support the notion that heparin may provide a local protective function. A similar but much larger effect was also observed in the presence of the beta subunit only...

  11. Role of casein phosphopeptide amorphous calcium phosphate in remineralization of white spot lesions and inhibition of Streptococcus mutans?

    OpenAIRE

    Ruchi Vashisht; Rajamani Indira; Ramachandran, S.; Anil Kumar; Manali Ramakrishnan Srinivasan

    2013-01-01

    Introduction: To promote the remineralization by ionic exchange mechanism instead of invasive techniques many remineralizing agents can be used. Objective: To evaluate the remineralization effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSLs) and its inhibitory effect on Streptococcus mutans colonization. Materials and Methods: The study group consisted of 60 subjects exhibiting at least 1-WSL. Subjects were randomly divided into 2 groups:...

  12. Evaluation of remineralization capacity of casein phosphopeptide-amorphous calcium phosphate on the carbamide peroxide treated enamel

    OpenAIRE

    Penumatsa, Narendra Varma; Kaminedi, Raja Rajeswari; Baroudi, Kusai; Barakath, Ola

    2015-01-01

    Objective: The aim of this study was to evaluate the potential of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) in remineralizing the bleached enamel surface using micro-hardness. Materials and Methods: Thirty human enamel slabs were randomly divided into three groups (n = 10). Groups A and B were exposed to 20% carbamide peroxide and 35% carbamide peroxide gel, respectively. After the exposure to the bleaching agent, the slabs were kept in artificial saliva for 1-week. Group C ...

  13. Casein kinase II phosphorylates the eukaryote-specific C-terminal domain of topoisomerase II in vivo.

    OpenAIRE

    Cardenas, M E; Dang, Q; Glover, C V; Gasser, S M

    1992-01-01

    The decatenation activity of DNA topoisomerase II is essential for viability as eukaryotic cells traverse mitosis. Phosphorylation has been shown to stimulate topoisomerase II activity in vitro. Here we show that topoisomerase II is a phosphoprotein in yeast and that the level of incorporated phosphate is significantly higher at mitosis than in G1. Comparison of tryptic phosphopeptide maps reveals that the major phosphorylation sites in vivo are targets for casein kinase II. Incorporation of ...

  14. Effect of casein phosphopeptide-amorphous calcium phosphate and acidulated phosphate fluoride gel on erosive enamel wear

    OpenAIRE

    Maryam HajeNorouz Ali Tehrani; Maryam Ghafournia; Pouran Samimi; Omid Savabi; Iman Parisay; Navid Askari; Seyed-Hossein Abtahi

    2011-01-01

    Background: Some studies have shown that casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and acidulated phosphate fluoride (APF) gel can protect teeth against erosion. The aim of this study was to assess whether CPP-ACP and fluoride could reduce enamel wear rates under erosive conditions simulating abrasion and acidic diet regimen. Materials and Methods: Enamel specimens consisted of 3 experimental groups (receiving CPP-ACP, APF or both) and a control group. Specimens were sub...

  15. Effect of casein phosphopeptide-amorphous calcium phosphate and acidulated phosphate fluoride gel on erosive enamel wear

    OpenAIRE

    Tehrani, Maryam HajeNorouz Ali; Ghafournia, Maryam; Samimi, Pouran; Savabi, Omid; Parisay, Iman; Askari, Navid; Abtahi, Seyed-Hossein

    2011-01-01

    Background: Some studies have shown that casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and acidulated phosphate fluoride (APF) gel can protect teeth against erosion. The aim of this study was to assess whether CPP-ACP and fluoride could reduce enamel wear rates under erosive conditions simulating abrasion and acidic diet regimen. Materials and Methods: Enamel specimens consisted of 3 experimental groups (receiving CPP-ACP, APF or both) and a control group. Specimens were subject...

  16. Efficacy of casein phosphopeptide-amorphous calcium phosphate to prevent stain absorption on freshly bleached enamel: An in vitro study

    OpenAIRE

    Raghuwar D Singh; Ram, Sabita M; Shetty, Omkar; Chand, Pooran; Yadav, Rakesh

    2010-01-01

    Background: Teeth when subjected to bleaching bring about the desiccation of the enamel, making it more susceptible to stain absorption. While subjecting the freshly bleached enamel surface to various surface treatments of Fluoride and Casein Phosphopeptide - Amorphous Calcium phosphate (CPP-ACP) brought about the reduction in stain absorption, which is assessed in this study. Aims: The study aims to evaluate the tea stain absorption on freshly bleached enamel surface of extracted human teeth...

  17. The role of casein phosphopeptide-amorphous calcium phosphate products in remineralization of incipient enamel lesions and its substantivity

    OpenAIRE

    Abhay Taranath; Deepika Pai; Kalyan Chakravarthy

    2014-01-01

    Objective: Milk derived calcium, phosphate and casein were known to be anti-cariogenic but in the modern era of early interception of caries the agents like casien phosphopeptide-amorphous calcium phosphate (CPP-ACP) derived from milk are known to also repair early lesions. This in vitro study was designed to evaluate the remineralizing potential of CPP-ACP and CPP-amorphous calcium phosphate fluoride (ACPF) on smooth surface carious lesions of the enamel and its substantivity following demin...

  18. The Effects of A Gluten and Casein-free Diet in Children with Autism: A Case Report

    OpenAIRE

    Chia-Lin Hsu; Delmar C.Y. Lin; Chia-Lin Chen; Chin-Man Wang; Wong, Alice M. K.

    2009-01-01

    A boy with autism, growth and developmental retardation was brought to our clinic. Hewas diagnosed with CHARGE syndrome. Subsequently, various therapies were introducedwhen he was 5 months old yet the developmental delays persisted. Gastrointestinal problemssuch as frequent post-prandial vomiting and severe constipation were noted as well. At theage of 42 months, the boy was subjected to a gluten and casein-free diet. Soybean milk andrice were substituted for cow’s milk, bread and noodles. Af...

  19. Conformational changes to deamidated wheat gliadins and b-casein upon adsorption to oil-water emulsion interfaces

    DEFF Research Database (Denmark)

    Wong, Benjamin T.; Zhai, Jiali; Hoffmann, Søren Vrønning;

    2012-01-01

    The conformation of deamidated gliadins and b-casein in solution and adsorbed at the interface of oil-inwater emulsions was studied using synchrotron radiation circular dichroism (SRCD) and front-facefluorescence spectroscopy. Deamidation led to partial unfolding of gliadins in solution. The a...... adsorbed state. Our results demonstrate that conformational re-arrangement of proteins upon adsorption to emulsion interfaces is dependent not only on the hydrophobicity of the oil phase, but more importantly on the conformational flexibility of the protein....

  20. Casein Kinase 2 Associates with Initiation-Competent RNA Polymerase I and Has Multiple Roles in Ribosomal DNA Transcription

    OpenAIRE

    Panova, Tatiana B; Panov, Kostya I.; Russell, Jackie; Zomerdijk, Joost C. B. M.

    2006-01-01

    Mammalian RNA polymerase I (Pol I) complexes contain a number of associated factors, some with undefined regulatory roles in transcription. We demonstrate that casein kinase 2 (CK2) in human cells is associated specifically only with the initiation-competent Pol Iβ isoform and not with Pol Iα. Chromatin immunoprecipitation analysis places CK2 at the ribosomal DNA (rDNA) promoter in vivo. Pol Iβ-associated CK2 can phosphorylate topoisomerase IIα in Pol Iβ, activator upstream binding factor (UB...

  1. The effect of two types chewing gum containing casein phosphopeptide-amorphous calcium phosphate and xylitol on salivary Streptococcus mutans

    OpenAIRE

    Shila Emamieh; Yosra Khaterizadeh; Hossein Goudarzi; Amir Ghasemi; Alireza Akbarzadeh Baghban; Hasan Torabzadeh

    2015-01-01

    Aim: The aim was to evaluate the effect of sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and xylitol on salivary Streptococcus mutans. Materials and Methods: A total of 60 dental students of 20-25 years old, who volunteered after checking their health condition and signing an informed consent, were randomly allocated to receive one of the following interventions: (A) Chewing gum containing CPP-ACP; (B) containing xylitol. Subjects within the ...

  2. Storage and lyophilization effects of extracts of Cynara cardunculus on the degradation of ovine and caprine caseins

    OpenAIRE

    Tavaria, Freni K.; Sousa, M. José; Malcata, F. Xavier

    2001-01-01

    Clotting and proteolytic activities are important parameters when evaluating rennets for cheesemaking. Both these activities were determined for extracts of the plant Cynara cardunculus in fresh form and after lyophilization followed by reconstitution, either in water or in citrate buffer (pH 5.4) and stored for up to 4 weeks at 4°C. The patterns of degradation of ovine and caprine caseins were followed by urea polyacrylamide gel electrophoresis in attempts to qualitatively differentiate the ...

  3. Exploring the physicochemical basis of cheese texture, rheology and functionality, with emphasis on cation-casein interactions

    OpenAIRE

    Cooke, Darren Richard

    2014-01-01

    The physicochemical properties of cheese and milk gels are greatly influenced by molecular interactions between the casein proteins involving calcium. Novel experiments were designed to investigate the relationship between insoluble caseinbound cations and rheological properties of Cheddar cheese and rennet-induced milk gels. Cheddar cheese and rennet-induced milk gels were supplemented with Mg2+ or Sr2+ to compare their effects on their rheological properties to those previously reported in ...

  4. Epigenetic effects of casein-derived opioid peptides in SH-SY5Y human neuroblastoma cells

    OpenAIRE

    Trivedi, Malav S.; Hodgson, Nathaniel W; Walker, Stephen J.; Trooskens, Geert; Nair, Vineeth; Deth, Richard C.

    2015-01-01

    Background Casein-free, gluten-free diets have been reported to mitigate some of the inflammatory gastrointestinal and behavioral traits associated with autism, but the mechanism for this palliative effect has not been elucidated. We recently showed that the opioid peptide beta-casomorphin-7, derived from bovine (bBCM7) milk, decreases cysteine uptake, lowers levels of the antioxidant glutathione (GSH) and decreases the methyl donor S-adenosylmethionine (SAM) in both Caco-2 human GI epithelia...

  5. Epigenetic effects of casein-derived opioid peptides in SH-SY5Y human neuroblastoma cells

    OpenAIRE

    Trivedi, Malav S.; Hodgson, Nathaniel W; Walker, Stephen J.; Trooskens, Geert; Nair, Vineeth; Deth, Richard C.

    2015-01-01

    Background: Casein-free, gluten-free diets have been reported to mitigate some of the inflammatory gastrointestinal and behavioral traits associated with autism, but the mechanism for this palliative effect has not been elucidated. We recently showed that the opioid peptide beta-casomorphin-7, derived from bovine (bBCM7) milk, decreases cysteine uptake, lowers levels of the antioxidant glutathione (GSH) and decreases the methyl donor S-adenosylmethionine (SAM) in both Caco-2 human GI epitheli...

  6. Identification of a BET family Bromodomain / Casein Kinase II / TAF-containing complex as a regulator of mitotic condensin function

    OpenAIRE

    Kim, Hyun-Soo; Mukhopadhyay, Rituparna; Rothbart, Scott B.; Silva, Andrea C.; Vanoosthuyse, Vincent; Radovani, Ernest; Kislinger, Thomas; Roguev, Assen; Ryan, Colm J.; Xu, Jiewei; Jahari, Harlizawati; Hardwick, Kevin G.; Greenblatt, Jack F.; Krogan, Nevan J.; Fillingham, Jeffrey S.

    2014-01-01

    Condensin is a central regulator of mitotic genome structure, with mutants showing poorly condensed chromosomes and profound segregation defects. Here we identify NCT complex, comprising the Nrc1 BET-family tandem bromodomain protein (SPAC631.02), Casein Kinase II (CKII) and several TAFs, as a regulator of condensin function. We show that NCT and condensin bind similar genomic regions, but only briefly co-localize during the periods of chromosome condensation and decondensation. This pattern ...

  7. The Gluten-Free/Casein-Free Diet: A Double-Blind Challenge Trial in Children with Autism

    Science.gov (United States)

    Hyman, Susan L.; Stewart, Patricia A.; Foley, Jennifer; Cain, Usa; Peck, Robin; Morris, Danielle D.; Wang, Hongyue; Smith, Tristram

    2016-01-01

    To obtain information on the safety and efficacy of the gluten-free/casein-free (GFCF) diet, we placed 14 children with autism, age 3-5 years, on the diet for 4-6 weeks and then conducted a double-blind, placebo-controlled challenge study for 12 weeks while continuing the diet, with a 12-week follow-up. Dietary challenges were delivered via weekly…

  8. Impact of cross-linking and drying method on drug delivery performance of casein-pectin microparticles.

    Science.gov (United States)

    Marreto, Ricardo N; Ramos, Monica F S; Silva, Emmanuelle J; de Freitas, Osvaldo; de Freitas, Luís A P

    2013-09-01

    Pectin is a heteropolysaccharide which has been investigated for the development of colon-specific drug delivery systems. Polymers have been associated with pectin to reduce its aqueous solubility and improve the performance of drug delivery systems. Pectin-casein interaction is widely known in food research, but it has not been fully considered by pharmaceutical scientists. Thus, this study investigated the potential of casein-pectin microparticles as a drug delivery system and clarified the impact of cross-linking and drying methods on the in vitro release of indomethacin (IND) or acetaminophen (PCT) from microparticles. Microparticles were prepared by coacervation and dried by spray or spouted bed methods. Drug recovery, in vitro drug release, size, morphology, and the thermal and diffractometric properties of dried microparticles were determined. Spray-dried non-cross-linked microparticles were able to prolong IND release, and pectin was still degraded by pectinolytic enzymes. On the other hand, glutaraldehyde cross-linking prevented the enzymatic breakdown of pectin without improving IND release. Spouted bed drying reduced IND recovery from all microparticles when compared with spray drying, thus the successful spouted bed drying of microparticles depends on the chemical characteristics of both the drug and the polymer. Release data from PCT microparticles suggested that the microparticle formulation should be improved to bring about a more efficient delivery of water-soluble drugs. In conclusion, casein-pectin microparticles show great potential as a drug delivery system because casein reduces the water solubility of pectin. The drying method and cross-linking process had significant effects on the in vitro performance of these microparticles. PMID:23934432

  9. Crosslinking with transglutaminase does not change metabolic effects of sodium caseinate in model beverage in healthy young individuals

    Directory of Open Access Journals (Sweden)

    Juvonen Kristiina R

    2012-06-01

    Full Text Available Abstract Background Postprandial metabolic and appetitive responses of proteins are dependent on protein source and processing technique prior to ingestion. Studies on the postprandial effects of enzymatic crosslinking of milk proteins are sparse. Our aim was to study the effect of transglutaminase (TG-induced crosslinking of sodium caseinate on postprandial metabolic and appetite responses. Whey protein was included as reference protein. Methods Thirteen healthy individuals (23.3 ± 1.1 y, BMI 21.7 ± 0.4 kg/m2 participated in a single-blind crossover design experiment in which the subjects consumed three different isovolumic (500 g pourable beverages containing either sodium caseinate (Cas, 29 g, TG-treated sodium caseinate (Cas-TG, 29 g or whey protein (Wh, 30 g in a randomized order. Blood samples were collected at baseline and for 4 h postprandially for the determination of plasma glucose, insulin and amino acid (AA concentrations. Gastric emptying (GE was measured using the 13 C-breath test method. Appetite was assessed using visual analogue scales. Results All examined postprandial responses were comparable with Cas and Cas-TG. The protein type used in the beverages was reflected as differences in plasma AA concentrations between Wh and Cas, but there were no differences in plasma glucose or insulin responses. A tendency for faster GE rate after Wh was detected. Appetite ratings or subsequent energy intake did not differ among the protein beverages. Conclusions Our results indicate that the metabolic responses of enzymatically crosslinked and native sodium caseinate in a liquid matrix are comparable, suggesting similar digestion and absorption rates and first pass metabolism despite the structural modification of Cas-TG.

  10. The Effect of Β-casein Nanoparticles on Bioavailability and Cellular Uptake of Platinum Complex as a Cancer Drug

    Directory of Open Access Journals (Sweden)

    M Razmi

    2013-12-01

    Full Text Available Abstract Background & aim: Due to the low solubility and high toxicity of drugs, treatment of cancers is problematic therefore, the encapsulation and targeted delivery of therapeutic effect is required. The aim of this study was to investigate the effect of nanoparticles on cellular uptake and bioavailability of beta-casein on platinum complexes as cancer drugs. Methods: In the present experimental study, the physicochemical properties of nanoparticles as drug carriers of beta-casein devices using dynamic light scattering (DLS and scanning electron microscopy (SEM were investigated. In order to evaluate the toxicity effects of platinum complexes, the colon cancer cells in the absence or presence of free platinum complex concentration and nanoparticle loaded with platinum complexes were incubated for 24 and 48 hours. LD50 Values (concentration of compound causing 50% mortality in the cells was determined using the MTT assay. Data were analyzed by ANOVA and post-hoc test. Results: At a concentration of 1 mg ml, beta-casein nanoparticle drug carriers were synthesized in the range of 100 to 300 µM. In addition, the mortality rate in cancer cells by the release of platinum complexes (without and with the capsule, were 70 and 26 in 24 hours, and 60 µM and 21 µM in 48 hours respectively, Conclusion: The study showed that the bioavailability of the encapsulated platinum complexes increases and new drug delivery system may be a good candidate for the treatment of cancer. Key words: Beta-casein, Pt (II Complex, Bioavalibility, Nanocarrier, Micelle

  11. Isolation and sequencing of cDNA clones encoding alpha and beta subunits of Drosophila melanogaster casein kinase II.

    OpenAIRE

    Saxena, A.; Padmanabha, R; Glover, C V

    1987-01-01

    Cloned cDNAs encoding both subunits of Drosophila melanogaster casein kinase II have been isolated by immunological screening of lambda gt11 expression libraries, and the complete amino acid sequence of both polypeptides has been deduced by DNA sequencing. The alpha cDNA contained an open reading frame of 336 amino acid residues, yielding a predicted molecular weight for the alpha polypeptide of 39,833. The alpha sequence contained the expected semi-invariant residues present in the catalytic...

  12. The Effect of Β-casein Nanoparticles on Bioavailability and Cellular Uptake of Platinum Complex as a Cancer Drug

    OpenAIRE

    M Razmi; A Divsalar

    2013-01-01

    Abstract Background & aim: Due to the low solubility and high toxicity of drugs, treatment of cancers is problematic therefore, the encapsulation and targeted delivery of therapeutic effect is required. The aim of this study was to investigate the effect of nanoparticles on cellular uptake and bioavailability of beta-casein on platinum complexes as cancer drugs. Methods: In the present experimental study, the physicochemical properties of nanoparticles as drug carriers of beta-ca...

  13. Wertigkeit von A1- und A2-Antikörpern gegen ß-Casein beim Typ 1-Diabetes mellitus

    OpenAIRE

    Seebaum, Silvia

    1998-01-01

    Im Zusammenhang mit dem insulinpflichtigen juvenilen Diabetes mellitus Typ 1 gewinnen in den letzten Jahren zunehmend Änderungen der Ernährungsgewohnheiten an Bedeutung. In zahlreichen Studien wurden verschiedene Bestandteile der Kuhmilch als möglicher Umweltfaktor bezüglich der Pathogenese des Typ 1-Diabetes mellitus untersucht. Für das Milchprotein Casein lagen bisher noch keine größeren Untersuchungen zur Wertigkeit hinsichtlich Ätiologie, Prävalenz und Pathogenese des Diabetes mellitu...

  14. Effect of kappa-casein and beta-lactoglobulin loci on milk production traits and reproductive performance of Holstein cows

    OpenAIRE

    Tsiaras, AM; Bargouli, GG; Banos, G.; Boscos, CM

    2005-01-01

    The effects of kappa-casein (kappa-CN) and beta-lactoglobulin (beta-LG) loci on milk production traits (milk, fat, protein, and lactose yield, fat, protein, and lactose content) and reproductive performance (gestation length, calving interval, age at first and second calving, number of services per conception) was estimated for 278 Holstein cows in the first 2 lactations. Genotypes of kappa-CN and beta-LG were determined by alkaline and acidic polyacrylamide gel electrophoresis. Milk producti...

  15. METHOD OF OBTAINING MICROPARTICULATED CASEIN AND THE POSSIBILITY OF ITS APPLICATION IN THE PRODUCTION OF NONFAT FERMENTED MILK PRODUCTS

    OpenAIRE

    Smirnova, I; Romanovskaya, I.; Shtrigul, V.

    2013-01-01

    Reducing the calorie content of everyday food, including dairy beverages, is among the key recommendations of present-day dietitians. However, fat reduction worsens the organoleptically evaluated texture and flavor score of food. The organoleptic correction of nonfat products using fat simulants often needs additional financial inputs. Furthermore, fat simulants added to the product can be hazardous to human health. Taking into consideration these facts, we have developed a casein micropartic...

  16. Caseinophosphopeptides released after tryptic hydrolysis versus simulated gastrointestinal digestion of a casein-derived by-product.

    Science.gov (United States)

    Cruz-Huerta, E; García-Nebot, M J; Miralles, B; Recio, I; Amigo, L

    2015-02-01

    The production of caseinophosphopeptides from a casein-derived by-product generated during the manufacture of a functional ingredient based on antihypertensive peptides was attempted. The casein by-product was submitted to tryptic hydrolysis for 30, 60 and 120min and further precipitated with calcium chloride and ethanol at pH 4.0, 6.0 and 8.0. Identification and semi quantification of the derived products by tandem mass spectrometry revealed some qualitative and quantitative changes in the released caseinophosphopeptides over time at the different precipitation pHs. The by-product was also subjected to simulated gastrointestinal digestion. Comparison of the resulting peptides showed large sequence homology in the phosphopeptides released by tryptic hydrolysis and simulated gastrointestinal digestion. Some regions, specifically αS1-CN 43-59, αS1-CN 60-74, β-CN 1-25 and β-CN 30-50 showed resistance to both tryptic hydrolysis and simulated digestion. The results of the present study suggest that this casein-derived by-product can be used as a source of CPPs. PMID:25172759

  17. pH-driven encapsulation of curcumin in self-assembled casein nanoparticles for enhanced dispersibility and bioactivity.

    Science.gov (United States)

    Pan, Kang; Luo, Yangchao; Gan, Yundi; Baek, Seung J; Zhong, Qixin

    2014-09-21

    The poor water solubility and bioactivity of lipophilic phytochemicals can be potentially improved by delivery systems. In this study, a low-cost, low-energy, and organic solvent-free encapsulation technology was studied by utilizing the pH-dependent solubility properties of curcumin and self-assembly properties of sodium caseinate (NaCas). Curcumin was deprotonated and dissolved, while NaCas was dissociated at pH 12 and 21 °C for 30 min. The subsequent neutralization enabled the encapsulation of curcumin in self-assembled casein nanoparticles. The degradation of curcumin under encapsulation conditions was negligible based on visible light and nuclear magnetic resonance spectroscopy. The dissociation of NaCas at pH 12 and reassociation after neutralization were confirmed using dynamic light scattering and analytical ultracentrifugation. The curcumin encapsulated in casein nanoparticles showed significantly improved anti-proliferation activity against human colorectal and pancreatic cancer cells. The studied encapsulation method is promising to utilize lipophilic compounds in food or pharmaceutical industries. PMID:25082426

  18. Real-time monitoring of peptic and tryptic digestions of bovine {beta}-casein using quartz crystal microbalance

    Energy Technology Data Exchange (ETDEWEB)

    Huenerbein, Andreas [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany)]. E-mail: andreas.huenerbein@pharmazie.uni-halle.de; Schmelzer, Christian E.H. [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany); Neubert, Reinhard H.H. [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany)

    2007-02-12

    In this study peptic and tryptic digestions of bovine {beta}-casein were investigated using quartz crystal microbalance (QCM). {beta}-Casein, which was used as a model protein, was immobilized on the surface of the QCM sensor where its degradation caused shifts in the resonant frequency. Atomic force microscopy was applied for the characterization of the protein layer. Different pH-values for peptic or tryptic digestions were chosen to visualize their effect on enzyme activity. Lower frequency shifts were observed at pH-values deviating from those at the maximum enzyme activity. In the case of the peptic digestion the frequency shift at pH 4 was more than 10 times smaller than those at pH 2. The frequency shifts for tryptic digestions at pH 5.4 and pH 6.4 were about two thirds compared to that obtained for the digestion at pH 7.4. The identification of peptides using MALDI-ToF mass spectrometry was used for verification of the proteolyses of the immobilized protein. Furthermore, it was shown that the QCM technique allows close observation of the effect of different pH-values on the immobilized casein layer. All in all, QCM facilitates the monitoring of the progress of enzymatic reactions in real-time.

  19. Prolactin and glucocorticoid signaling induces lactation-specific tight junctions concurrent with β-casein expression in mammary epithelial cells.

    Science.gov (United States)

    Kobayashi, Ken; Tsugami, Yusaku; Matsunaga, Kota; Oyama, Shoko; Kuki, Chinatsu; Kumura, Haruto

    2016-08-01

    Alveolar mammary epithelial cells (MECs) in mammary glands are highly specialized cells that produce milk for suckling infants. Alveolar MECs also form less permeable tight junctions (TJs) to prevent the leakage of milk components after parturition. In the formation process of less permeable TJs, MECs show a selective downregulation of Cldn4 and a localization change of Cldn3. To investigate what induces less permeable TJs through these compositional changes in Cldns, we focused on two lactogenesis-related hormones: prolactin (Prl) and glucocorticoids. Prl caused a downregulation of Cldn3 and Cldn4 with the formation of leaky TJs in MECs in vitro. Prl-treated MECs also showed low β-casein expression with the activation of STAT5 signaling. By contrast, dexamethasone (Dex), a glucocorticoid analogue, upregulated Cldn3 and Cldn4, concurrent with the formation of less permeable TJs and the activation of glucocorticoid signaling without the expression of β-casein. Cotreatment with Prl and Dex induced the selective downregulation of Cldn4 and the concentration of Cldn3 in the region of TJs concurrent with less permeable TJ formation and high β-casein expression. The inhibition of Prl secretion by bromocriptine in lactating mice induced the upregulation of Cldn3 and Cldn4 concurrent with the downregulation of milk production. These results indicate that the coactivation of Prl and glucocorticoid signaling induces lactation-specific less permeable TJs concurrent with lactogenesis. PMID:27130254

  20. Charge properties of peptides derived from casein affect their bioavailability and cytoprotection against H2O2-induced oxidative stress.

    Science.gov (United States)

    Wang, Bo; Xie, Ningning; Li, Bo

    2016-04-01

    The effects of charge properties of casein peptides on absorption stability, antioxidant activity, and cytoprotection were evaluated. Alcalase hydrolysates of casein were separated into 4 fractions by cation-exchange chromatography according to charge properties. After simulated digestion and Caco-2 cell transmembrane transport, we determined the total antioxidant capacity (Trolox equivalent antioxidative capacity and oxygen radical antioxidant activity) and nitrogen content of peptide fractions to estimate available antioxidant efficacy and bioavailability (BA) of peptides. Results showed that negatively charged peptide fractions had greater BA and antioxidant activities after digestion and absorption. The peptide permeates were used to test the cytoprotective effect against H2O2-induced oxidative damage in HepG-2 cells. All peptide permeates increased cell viability, elevated catalase activity, and decreased superoxide dismutase activity. However, negatively charged peptide fractions preserved cell viability to a greater degree. Therefore, the negatively charged peptides from casein may be potential antioxidants and could be used as ingredients in functional foods and dietary supplements. PMID:26851854

  1. The influence of casein haplotype on quality, coagulation, and yield traits of milk from Italian Holstein cows.

    Science.gov (United States)

    Perna, Annamaria; Intaglietta, Immacolata; Gambacorta, Emilio; Simonetti, Amalia

    2016-05-01

    The aim of this work was to investigate the effect of casein haplotype (CSN1S1, CSN2, and CSN3) on quality, coagulation, and yield traits of milk from Italian Holstein cows. The casein haplotype was determined by isoelectric focusing; milk clotting properties were determined by using a mechanical lacto-dynamographic instrument; and the yields of pressed and pasta filata cheeses were expressed as kilograms of cheese per 100kg of milk processed. Statistical analysis showed a significant effect of the casein haplotype. In particular, BB-A(1)A(1)-AA milk showed the highest fat content (4.01g/100g), whereas BB-A(2)A(2)-BB milk had a higher protein content, the best coagulation characteristics, and the highest yields in pressed and pasta filata cheeses, and, consequently, better ability to retain fat and protein in the curd. The results of this study suggest that knowledge of milk protein polymorphisms not only allows the production of milk with specific qualitative and quantitative characteristics, but it could also be used as a specific marker within a breed to identify milk suitable for cheesemaking, which confers an economical advantage for dairy producers. PMID:26947288

  2. Diffusion and partitioning of macromolecules in casein microgels: evidence for size-dependent attractive interactions in a dense protein system.

    Science.gov (United States)

    Peixoto, Paulo D S; Bouchoux, Antoine; Huet, Sébastien; Madec, Marie-Noëlle; Thomas, Daniel; Floury, Juliane; Gésan-Guiziou, Geneviève

    2015-02-10

    Understanding the mechanisms that determine the diffusion and interaction of macromolecules (such as proteins and polysaccharides) that disperse through dense media is an important fundamental issue in the development of innovative technological and medical applications. In the current work, the partitioning and diffusion of macromolecules of different sizes (from 4 to 10 nm in diameter) and shapes (linear or spherical) within dispersions of casein micelles (a protein microgel) is studied. The coefficients for diffusion and partition are measured using FRAP (fluorescence recovery after photobleaching) and analyzed with respect to the structural characteristics of the microgel determined by the use of TEM (transmission electron microscopy) tomography. The results show that the casein microgel displays a nonspecific attractive interaction for all macromolecules studied. When the macromolecular probes are spherical, this affinity is clearly size-dependent, with stronger attraction for the larger probes. The current data show that electrostatic effects cannot account for such an attraction. Rather, nonspecific hydration molecular forces appear to explain these results. These findings show how weak nonspecific forces affect the diffusion and partitioning of proteins and polysaccharides in a dense protein environment. These results could be useful to better understand the mechanisms of diffusion and partitioning in other media such as cells and tissues. Furthermore, there arises the possibility of using the casein micelle as a size-selective molecular device. PMID:25604622

  3. Effect of high pressure--low temperature treatments on structural characteristics of whey proteins and micellar caseins.

    Science.gov (United States)

    Baier, Daniel; Purschke, Benedict; Schmitt, Christophe; Rawel, Harshadrai M; Knorr, Dietrich

    2015-11-15

    In this study, structural changes in micellar caseins and whey proteins due to high pressure--low temperature treatments (HPLT) were investigated and compared to changes caused by high pressure treatments at room temperature. Whey protein isolate (WPI) solutions as well as micellar casein (MC) dispersions and mixtures were treated at 500 MPa (pH 7.0 and 5.8) at room temperature, -15 °C and -35 °C. Surface hydrophobicity and accessible thiol groups remained nearly unchanged after HPLT treatments whereas HP treatments at room temperature caused an unfolding of the WPI, resulting in an increase in surface hydrophobicity and exposure of the thiol groups. For HPLT treatments, distinct changes in the secondary structure (increase in the amount of β-sheets) were observed while the tertiary structure remained unchanged. Large flocs, stabilized by hydrophobic interactions and hydrogen bonds, were formed in casein containing samples due to HPLT treatments. Depending on the pH and the applied HPLT treatment parameters, these interactions differed significantly from the interactions determined in native micelles. PMID:25977037

  4. Gluten-free and casein-free diets in the treatment of autism

    Directory of Open Access Journals (Sweden)

    Andreas Reissmann

    2014-08-01

    Full Text Available Background: Autism is a complex psychiatric disorder characterized by three core symptoms, i.e. impairments in social interaction, restricted patterns of behavior and impairments in communication. In the framework of the “opioid excess theory”, the disorder symptoms are compared to the behavioral effects of opiates. Based on this, a possible nutritional basis of autism has been proposed, hypothesizing that certain food proteins such as gluten and casein can be transformed to opioid peptides during digestion. These peptides might eventually be able to enter the blood stream and act upon the central nervous system. As a consequence, a diet low in such proteins has been hypothesized to ameliorate the behavioral symptoms of autistic children. Objective: The scope of this review was to analyze the effects of gluten-free and casein-free (GFCF diets on children with autism, as well as to provide information concerning additional aspects related to the GFCF diet in autism. Methods: A literature search was conducted including scientific publications up until December 2013. Search results were screened for any kind of GFCF dietary intervention as well as surveys dealing with GFCF as a treatment for autism. Results: A review of survey data shows that up to 25 % of parents of affected children report on current use of a GFCF diet. The majority of identified studies evaluating GFCF diet outcomes failed to meet basic methodological standards of interventional science. Comparison of study results did not show any clear-cut results, with a substantial proportion of studies failing to show any positive dietary effect. The results of more sophisticated trials were far from equivocal and the studies differed by many methodological aspects. Some variables such as information source and trial duration seemed to affect outcome. Conclusions: Evidence for the effectiveness of the GFCF diet in the treatment of autism is sparse. Rigorous scientific evaluations partly

  5. A review of gluten- and casein-free diets for treatment of autism: 2005–2015

    Directory of Open Access Journals (Sweden)

    Elder JH

    2015-12-01

    Full Text Available Jennifer Harrison Elder,1 Consuelo Maun Kreider,2 Nancy M Schaefer,3 Mary B de Laosa4 1Department of Family and Community Health Nursing Science, 2Department of Occupational Therapy, 3Health Science Center Library, 4Department of Psychology, University of Florida, Gainesville, FL, USA Background: The gluten-free, casein-free (GFCF diet is heralded by strong anecdotal parental reports to greatly improve and even "cure" symptoms of autism spectrum disorders (ASDs. Yet, to date, little conclusive empirical evidence exists supporting its use. Objective: The purpose of this paper is to provide an overview of the state of the recent evidence regarding the use of GFCF diet for treatment of individuals with ASD. Methods: Five database providers (PubMed, Web of Knowledge, EBSCO, ProQuest, and WorldCat were used to search 19 databases, yielding a total of 491 articles that were published through February 2015. Peer-reviewed articles published between January 2005 and February 2015 were included for review if study participants were identified as having ASD and if the study investigated the effects of the GFCF diet on ASD behaviors or the relationship between the diet and these behaviors. Results: Evaluation of search results yielded eleven reviews, seven group experimental studies including five randomized controlled trials, five case reports, and four group observational studies published during the last 10 years. These studies represent a marked increase in the number of reported studies as well as increased scientific rigor in investigation of GFCF diets in ASD. Conclusion: While strong empirical support for the GFCF diet in ASD is currently lacking, studies point to the need for identifying subsets of individuals (eg, those with documented gastrointestinal abnormalities who may be the best responders to the GFCF diet. Identifying these subsets is critically needed to enhance rigor in this research area. Until rigorous research supporting the use of

  6. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion related to a notification from the International Organisation of Vine and Wine (OIV) on casein/caseinate/milk products to be used in the manufacture of wine as clarification processing aids pursuant to Article 6

    DEFF Research Database (Denmark)

    Tetens, Inge

    wine as clarification processing aids pursuant to Article 6, paragraph 11 of Directive 2000/13/EC – for permanent exemption from labelling. In earlier assessments, the Panel concluded that wines fined with casein/caseinate/milk products may trigger adverse reactions in susceptible individuals under the...... refers to new analytical methods developed for the detection of milk allergens in the fining agent and the detection of casein in wine. There were no changes in the wine manufacturing process and no new clinical studies were provided. Taking into account the information provided on the characterisation...

  7. Effects of milk containing only A2 beta casein versus milk containing both A1 and A2 beta casein proteins on gastrointestinal physiology, symptoms of discomfort, and cognitive behavior of people with self-reported intolerance to traditional cows’ milk

    OpenAIRE

    Jianqin, Sun; Leiming, Xu; Lu, Xia; Yelland, Gregory W; Ni, Jiayi; Clarke, Andrew J.

    2016-01-01

    Background Cows’ milk generally contains two types of β-casein, A1 and A2 types. Digestion of A1 type can yield the peptide β-casomorphin-7, which is implicated in adverse gastrointestinal effects of milk consumption, some of which resemble those in lactose intolerance. This study aimed to compare the effects of milk containing A1 β-casein with those of milk containing only A2 β-casein on inflammation, symptoms of post-dairy digestive discomfort (PD3), and cognitive processing in subjects wit...

  8. Polydiacetylene/triblock copolymer nanoblend applied as a sensor for micellar casein: A thermodynamic approach.

    Science.gov (United States)

    de Souza, Luana Cypriano; de Paula Rezende, Jaqueline; Pires, Ana Clarissa dos Santos; da Silva, Luis Henrique Mendes; da Silva, Maria do Carmo Hespanhol; Castrillon, Elkin Dario Castellon; de Andrade, Nélio José

    2016-04-15

    Polydiacetylene (PDA) and triblock copolymer nanoblends were synthesized to detect micellar casein (MC), the main milk protein and an indicator of milk quality. UV-Vis spectrum showed that MC induced blue-to-red transition in nanoblends. When nanoblends and MC were separated by dialysis membrane colorimetric response (CR) was similar, whereas a remarkable CR reduction was noticed after addition of dialyzed-MC, suggesting that small molecules present in MC (salts) caused PDA color change. Interaction enthalpy variation between nanoblends and MC showed an abrupt increase that coincided with MC concentration when colorimetric transition occurred. Copolymer hydrophobic/hydrophilic balance and presence of other molecules in the system affected nanoblends CR. MC salts were found to interact with nanoblends leading to color changes. MC concentration, MC salt release, copolymer hydrophobic/hydrophilic balance, and presence of other molecules in the system affected responses of the sensors. These results contribute to future applications of PDA/copolymer nanosensors to dairy models. PMID:26617025

  9. A PER2-derived mechanism-based bisubstrate analog for casein kinase 1ε.

    Science.gov (United States)

    Zeringo, Nicholas A; Bellizzi, John J

    2014-12-01

    Casein kinase 1ε (CK1ε) plays an important regulatory role in various cellular processes including circadian rhythms. Mutations in CK1ε or the recognition site on its substrate PER2 result in modulation of the circadian period length. In particular, the tau mutation (R178C) in the catalytic domain of CK1ε was identified as the molecular basis for a dose-dependent heritable shortened circadian period in hamsters. However, the biochemical basis for the physiological effects of the tau mutant remains unclear. It has been reported that the tau mutation has reduced in vitro activity against some substrates but increased in vitro activity against other substrates. To better understand the effects of the CK1ε tau mutation, an ATP-phosphopeptide conjugate was synthesized to yield a transition-state bisubstrate analog. Kinase activity assays determined that the tau mutant has 80% reduced activity and a fourfold decrease in sensitivity to the bisubstrate analog compared to wild type. This confirms that Arg178 is important in the recognition of the preferred phosphosubstrates of CK1ε. PMID:24985607

  10. Preparation and bioactive properties of chitosan and casein phosphopeptides composite coatings for orthopedic implants.

    Science.gov (United States)

    Qin, Liguo; Dong, Huanhuan; Mu, Ziqing; Zhang, Yali; Dong, Guangneng

    2015-11-20

    Using the layer-by-layer deposition method, functional chitosan/casein phospopeptides (CS/CPP) composite coatings were produced on Co-Cr-Mo alloy. The CS/CPP composite coatings had the dendritic topography, and were quite hydrophilic. Zeta potential measurements showed the composite coatings were negative charged at neural pH. XPS results indicated that the CS/CPP composite coatings were covalently bond to the substrate. When MC3T3-E1 cells were seeded on the CS/CPP composite coatings, no cytotoxicity was observed. The bone morphogenetic protein-2 (BMP-2) mRNA expression was significantly up-regulated in MC3T3-E1 cells cultured on the composite coatings and it was twice as much as that of cells cultured on the bare substrate. The expression of osteoprotegerin (OPG) mRNA and the ratio of OPG/receptor activator of nuclear factor-κB ligand (RNAKL) mRNA were increased 5-fold and 55-fold, respectively. These results suggested the CS/CPP composite coatings may have potential application in cobalt matrix orthopaedic implants. PMID:26344277

  11. REGγ deficiency promotes premature aging via the casein kinase 1 pathway.

    Science.gov (United States)

    Li, Lei; Zhao, Dengpan; Wei, Haibin; Yao, Liangfang; Dang, Yongyan; Amjad, Ali; Xu, Jinjin; Liu, Jiang; Guo, Linjie; Li, Dongqing; Li, Zhen; Zuo, Di; Zhang, Yuanyuan; Liu, Jian; Huang, Shixia; Jia, Caifeng; Wang, Lu; Wang, Ying; Xie, Yifan; Luo, Jian; Zhang, Bianhong; Luo, Honglin; Donehower, Lawrence A; Moses, Robb E; Xiao, Jianru; O'Malley, Bert W; Li, Xiaotao

    2013-07-01

    Our recent studies suggest a role for the proteasome activator REG (11S regulatory particles, 28-kDa proteasome activator)γ in the regulation of tumor protein 53 (p53). However, the molecular details and in vivo biological significance of REGγ-p53 interplay remain elusive. Here, we demonstrate that REGγ-deficient mice develop premature aging phenotypes that are associated with abnormal accumulation of casein kinase (CK) 1δ and p53. Antibody array analysis led us to identify CK1δ as a direct target of REGγ. Silencing CK1δ or inhibition of CK1δ activity prevented decay of murine double minute (Mdm)2. Interestingly, a massive increase of p53 in REGγ(-/-) tissues is associated with reduced Mdm2 protein levels despite that Mdm2 transcription is enhanced. Allelic p53 haplodeficiency in REGγ-deficient mice attenuated premature aging features. Furthermore, introducing exogenous Mdm2 to REGγ(-/-) MEFs significantly rescues the phenotype of cellular senescence, thereby establishing a REGγ-CK1-Mdm2-p53 regulatory pathway. Given the conflicting evidence regarding the "antiaging" and "proaging" effects of p53, our results indicate a key role for CK1δ-Mdm2-p53 regulation in the cellular aging process. These findings reveal a unique model that mimics acquired aging in mammals and indicates that modulating the activity of the REGγ-proteasome may be an approach for intervention in aging-associated disorders. PMID:23766372

  12. SOYBEAN AND CASEIN HYDROLYSATES INDUCE GRAPEVINE IMMUNE RESPONSES AND RESISTANCE AGAINST PLASMOPARA VITICOLA

    Directory of Open Access Journals (Sweden)

    Nihed eLachhab

    2014-12-01

    Full Text Available Plasmopara viticola, the causal agent of grapevine downy mildew, is one of the most devastating grape pathogen in Europe and North America. Although phytochemicals are used to control pathogen infections, the appearance of resistant strains and the concern for possible adverse effects on environment and human health are increasing the search for alternative strategies. In the present investigation, we successfully tested two protein hydrolysates from soybean (soy and casein (cas to trigger grapevine resistance against P. viticola. On Vitis vinifera cv. Marselan plants, the application of soy and cas reduced the infected leaf surface by 76 and 63%, as compared to the control, respectively. Since both hydrolysates might trigger the plant immunity, we investigated their ability to elicit grapevine defence responses. On grapevine cell suspensions, a different free cytosolic calcium signature was recorded for each hydrolysate, whereas a similar transient phosphorylation of two MAP kinases of 45 and 49 kDa was observed. These signalling events were followed by transcriptome reprogramming, including the up-regulation of defence genes encoding pathogenesis-related (PR proteins and the stilbene synthase enzyme responsible for the biosynthesis of resveratrol, the main grapevine phytoalexin. Liquid chromatography analyses confirmed the production of resveratrol and its dimer metabolites, δ- and ε-viniferins. Overall, soy effects were more pronounced as compared to the cas one. Both hydrolysates proved to act as elicitors to enhance grapevine immunity against pathogen attack.

  13. Casein Kinase 1 Regulates Ethylene Synthesis by Phosphorylating and Promoting the Turnover of ACS5

    Directory of Open Access Journals (Sweden)

    Shu-Tang Tan

    2014-12-01

    Full Text Available The casein kinase 1 (CK1 family participates in various cellular processes in eukaryotes, but CK1 function in higher plants remains largely unknown. Here, we characterize the function of Arabidopsis CK1 in the regulation of ethylene biosynthesis. Etiolated seedlings of a CK1.8-deficient mutant, ck1.8-1, showed characteristic ethylene-specific constitutive responses due to overaccumulation of ethylene. Biochemical and physiological studies showed that CK1.8 phosphorylates ACS5, a key enzyme of ethylene biosynthesis, at threonine 463 to promote its interaction with the E3 ubiquitin ligase Ethylene Overproduction 1 (ETO1. Deficiency of CK1.8 leads to the accumulation of ACS5, and transgenic plants harboring a dephosphorylation-mimic ACS5T463A showed constitutive ethylene responses, confirming the role of CK1.8 in regulating ACS5 stability by phosphorylation and demonstrating that CK1.8 is an important regulator of ethylene biosynthesis. CK1.8 expression is feedback regulated by ethylene. Our studies provide insight into the regulation of ACS5 and ethylene biosynthesis.

  14. Complex coacervation for the development of composite edible films based on LM pectin and sodium caseinate.

    Science.gov (United States)

    Eghbal, Noushin; Yarmand, Mohammad Saeid; Mousavi, Mohammad; Degraeve, Pascal; Oulahal, Nadia; Gharsallaoui, Adem

    2016-10-20

    Coacervation between sodium caseinate (CAS) and low methoxyl pectin (LMP) at pH 3 was investigated as a function of protein/polysaccharide ratio. The highest amount of complex coacervates was formed at a CAS/LMP ratio of 2 at which the ζ-potential value was zero and the turbidity reached its highest value. Then, the properties of films based on these complex coacervates were studied. Coacervation resulted in decreasing water content and water sorption of films as the protein concentration increased. The mechanical properties of films were highly influenced by the formation of electrostatic complexes. The highest values of Young's modulus (182.97± 6.48MPa) and tensile strength (15.64±1.74MPa) with a slight increase of elongation at break (9.35±0.10%) were obtained for films prepared at a CAS/LMP ratio equal to 0.05. These findings show that interactions between LMP and CAS can be used to develop innovative packaging containing active molecules. PMID:27474643

  15. Casein kinase 2 inhibits HomolD-directed transcription by Rrn7 in Schizosaccharomyces pombe.

    Science.gov (United States)

    Moreira-Ramos, Sandra; Rojas, Diego A; Montes, Matías; Urbina, Fabiola; Miralles, Vicente J; Maldonado, Edio

    2015-02-01

    In Schizosaccharomyces pombe, ribosomal protein gene (RPG) promoters contain a TATA analogue element called the HomolD box. The HomolD-binding protein Rrn7 forms a complex with the RNA polymerase II machinery. Despite the importance of ribosome biogenesis to cell survival, the mechanisms involved in the regulation of transcription of eukaryotic RPGs are unknown. In this study, we identified Rrn7 as a new substrate of the pleiotropic casein kinase 2 (CK2), which is a regulator of basal transcription. Recombinant Rrn7 from S. pombe, which is often used as a model organism for studying eukaryotic transcription, interacted with CK2 in vitro and in vivo. Furthermore, CK2-mediated phosphorylation of Rrn7 inhibited its HomolD-directed transcriptional activity and ability to bind to an oligonucleotide containing a HomolD box in vitro. Mutation of Rrn7 at Thr67 abolished these effects, indicating that this residue is a critical CK2 phosphorylation site. Finally, Rrn7 interacted with the regulatory subunit of CK2 in vivo, inhibition of CK2 in vivo potentiated ribosomal protein gene transcription, and chromatin immunoprecipitation analyses identified that the catalytic subunit of CK2 was associated with the rpk5 gene promoter in S. pombe. Taken together, these data suggest that CK2 inhibits ribosomal protein gene transcription in S. pombe via phosphorylation of Rrn7 at Thr67. PMID:25410910

  16. Casein kinase G may be the target of spermine during progesterone-induced oocyte maturation

    Institute of Scientific and Technical Information of China (English)

    LIRUNSHENG; JIAKETSO

    1993-01-01

    Casein kinase G (CKG) with more than 2500-fold enrichraent was purified from Bufo bufo gargarizans ovaries. The catalytic activity of the enzyme was found to be associated with its 42 kD subunit, and its 26 kD subunit was found to be the major tsrget for the enzyme autophos phorylation. Each fuU-grown oocyte contained 1.9 units of CKG corresponding to an intracellular concentration of 93 nM. After injecting an amount of 0,38 units of the enzyme into the oocyte, approximately 50% of the progesterone-induoed maturation was inhibited. The inhibitory effect was enhanced in oocytes pretreated with spermine, which was consistent with the results that the enzyme was activated in vitro in the presence of spermine, The MPF-induced oocyte maturation was delayed and even prohibited in the kinase-microinjected oocytes. A 55 kD oocyte protein was identified as an suhstrate of CKG both in vivo and in vitro, and the enhancement of the 55 kD protein phosphory[ation was associated with kinase inhibition on maturation and on protein synthesis in kinase-microinjected oocytes. As the endogenous spermine level decreased in the course of progesteroneinduced oocyte maturation. 55 kD protein was dephosphorylated, Heparin, a specific inhibitor of CKG, potentiated the progesterone-induced oocyte maturation. Altogether the experimental reSults indicated Strongly that CKG may be the physiological target of spermine.

  17. Casein Kinase 2: a novel player in glioblastoma therapy and cancer stem cells.

    Science.gov (United States)

    Agarwal, Maya; Nitta, Ryan T; Li, Gordon

    2013-12-01

    Casein kinase 2 (CK2) is an oncogenic protein kinase which contributes to tumor development, proliferation, and suppression of apoptosis in multiple cancer types. The mechanism by which CK2 expression and activity leads to tumorigenesis in glioblastoma (GBM), a stage IV primary brain tumor, is being studied. Recent studies demonstrate that CK2 plays an important role in GBM formation and growth through the inhibition of tumor suppressors and activation of oncogenes. In addition, intriguing new reports indicate that CK2 may regulate GBM formation in a novel manner; CK2 may play a critical role in cancer stem cell (CSC) maintenance. Since glial CSCs have the ability to self-renew and initiate tumor growth, new treatments which target these CSCs are needed to treat this fatal disease. Inhibition of CK2 is potentially a novel method to inhibit GBM growth and reoccurrence by targeting the glial CSCs. A new, orally available, selective CK2 inhibitor, CX-4945 has had promising results when tested in cancer cell lines, in vivo xenograft models, and human clinical trials. The development of CK2 targeted inhibitors, starting with CX-4945, may lead to a new class of more effective cancer therapies. PMID:25264454

  18. Influence of sodium chloride on the colloidal and rennet coagulation properties of concentrated casein micelle suspensions.

    Science.gov (United States)

    Zhao, Z; Corredig, M

    2016-08-01

    The research investigated the influence of NaCl on the colloidal and rennet coagulation properties of concentrated milk. Milk was concentrated to 1×, 3×, and 5× using ultrafiltration. Rennet gelation was followed by rheology and diffusing wave spectroscopy. Soluble protein, total and diffusible calcium and phosphate, size, and zeta potential were also measured as a function of concentration history. In the presence of 300mM NaCl, colloidal calcium phosphate solubilized and pH and the negative charge on the surface of casein micelles decreased. Increasing the volume fraction caused the formation of stiffer gels for both samples with or without NaCl. The addition of NaCl caused a significant increase in the bulk viscosity of the milk concentrated 5× and a decrease in turbidity. The concentration had no effect on the gelation time of control samples, nor on the kinetics of caseinomacropeptide release. On the other hand, rennet gelation was retarded by the addition of NaCl, and the gels showed lower elastic moduli compared with those obtained with control milk. PMID:27320668

  19. Casein Kinase 2 Reverses Tail-Independent Inhibition of Kinesin-1

    Science.gov (United States)

    Xu, Jing; Shu, Zhanyong; Anand, Preetha; Reddy, Babu; Cermelli, Silvia; Whisenant, Thomas; King, Stephen; Bardwell, Lee; Huang, Lan; Gross, Steven

    2011-03-01

    Kinesin-1 is a plus-end microtubule-based molecular motor, and defects in kinesin transport are linked to diseases including neurodegeneration. Kinesin can auto-inhibit via a direct head-tail interaction, but is believed to be active otherwise. In contrast, this study uncovers a fast but reversible inhibition distinct from the canonical auto-inhibition pathway. The majority of the initially active kinesin (full-length or tail-less) loses its ability to bind/interact with microtubule, and Casein Kinase 2 (CK2) reverses this inactivation (up to 4-fold) without altering kinesin's single motor properties. Motor phosphorylation is not required for this CK2 -mediated kinesin activation. In cultured mammalian cells, knockdown of CK2 level, but not kinase activity, was sufficient to decrease the force required to stall lipid droplet transport, consistent with a reduction in the number of active motors. We propose that CK2 forms a positive regulating complex with the motor. This study provides the first direct evidence of a protein kinase positively regulating kinesin-transport, and uncovers a pathway whereby inactive cargo-bound kinesin can be activated. This work is supported by NIGMS grants GM64624 and GM079156 to SPG, GM-74830 to LH, NIH grants GM76516 and GM60366 to LB, and AHA grant 825278F to JX.

  20. Casein Kinase 2 Reverses Tail-Independent Inactivation of Kinesin-1

    Science.gov (United States)

    Xu, Jing

    2013-03-01

    Kinesin-1 is a plus-end microtubule-based motor, and defects in kinesin-based transport are linked to diseases including neurodegeneration. Kinesin can auto-inhibit via a head-tail interaction, but is believed to be active otherwise. Here we report a tail-independent inactivation of kinesin, reversible by the disease-relevant signalling protein, casein kinase 2 (CK2). The majority of initially active kinesin (native or tail-less) loses its ability to interact with microtubules in vitro, and CK2 reverses this inactivation (approximately fourfold) without altering kinesin's single motor properties. This activation pathway does not require motor phosphorylation, and is independent of head-tail auto-inhibition. In cultured mammalian cells, reducing CK2 expression, but not its kinase activity, decreases the force required to stall lipid droplet transport, consistent with a decreased number of active kinesin motors. Our results (Nat. Commun., 3:754, 2012) provide the first direct evidence of a protein kinase upregulating kinesin-based transport, and suggest a novel pathway for regulating the activity of cargo-bound kinesin. Work supported by NIGMS grants GM64624 to SPG, GM74830-06A1 to LH, GM76516 to LB, NS048501 to SJK, and AHA grant 825278F to JX.

  1. Controllable self-assembly of sodium caseinate with a zwitterionic vitamin-derived bolaamphiphile.

    Science.gov (United States)

    Sun, Li-Hui; Sun, Yu-Long; Yang, Li-Jun; Zhang, Jian; Chen, Zhong-Xiu

    2013-11-01

    The control of self-assembly of sodium caseinate (SC) including the formation of mixed layers, microspheres, or nanoparticles is highly relevant to the microstructure of food and the design of promising drug delivery systems. In this paper, we designed a structure-switchable zwitterionic bolaamphiphile, 1,12-diaminododecanediorotate (DDO), from orotic acid, which has special binding sites and can guide the self-assembly of SC. Complexation between SC and DDO was investigated using dynamic light scattering, transmission electron microscopy, differential scanning calorimetry, and fluorescence spectra measurements. Monomeric DDO was bound to the negatively charged sites on the SC micelle and made the structure of SC more compact with decreased electrostatic repulsion between the head groups. Vesicular DDO led to reassociation of vesicles with enlarged size via preferable hydrophobic interactions. Moreover, the aggregation between SC and DDO was found to be temperature-dependent and reversible. This research provides an effective way to control the reversible self-assembly of SC by the zwitterionic vitamin-derived bolaamphiphile. PMID:24107219

  2. Effect of temperature and loading on the structure of β-casein/ibuprofen assemblies.

    Science.gov (United States)

    Turovsky, Tanya; Portnaya, Irina; Kesselman, Ellina; Ionita-Abutbul, Inbal; Dan, Nily; Danino, Dganit

    2015-07-01

    β-Casein is a 24 kDa amphiphilic and unstructured protein that self-assembles into small core-shell micelles at a wide range of concentrations, pH values and temperatures. We recently developed the micelles as nanocarriers for oral delivery of hydrophobic drugs. In this paper we examined the effect of the hydrophobic non-steroidal anti-inflammatory drug (NSAID) ibuprofen on the micellar structure, as a function of temperature and loading. Using cryo-transmission electron microscopy (cryo-TEM) we find two routes of organization – mixed micellization and co-assembly (aggregation). The time-dependent events that characterize the second routes has been examined in detail. At 25 °C we find coexistence of small assemblies and larger aggregates of irregular (but defined) structures that contain the drug. Increasing the drug loading increases the relative number of the larger aggregates and their dimensions, leading eventually to the formation of long then branched structures, like in amphiphilic block copolymer solutions. Similar trends were identified for changes in the temperature. Combined, our results suggest that ibuprofen acts as a co-surfactant that possibly is localizes to the interface rather than being encapsulated in the micellar core as other NSAID hydrophobic drugs. PMID:25754442

  3. Regulation of Ikaros function by casein kinase 2 and protein phosphatase 1

    Institute of Scientific and Technical Information of China (English)

    Amy; K; Erbe; Aleksandar; Savic; Sinisa; Dovat

    2011-01-01

    The Ikaros gene encodes a zinc finger,DNA-binding protein that regulates gene transcription and chromatin remodeling.Ikaros is a master regulator of hematopoiesis and an established tumor suppressor.Moderate alteration of Ikaros activity (e.g.haploinsufficiency) appears to be sufficient to promote malignant transformation in human hematopoietic cells.This raises questions about the mechanisms that normally regulate Ikaros function and the potential of these mechanisms to contribute to the development of leukemia.The focus of this review is the regulation of Ikaros function by phosphorylation/dephosphorylation.Site-specific phosphorylation of Ikaros by casein kinase 2 (CK2) controls Ikaros DNA-binding ability and subcellular localization.As a consequence,the ability of Ikaros to regulate cell cycle progression,chromatin remodeling,target gene expression,and thymocyte differentiation are controlled by CK2.In addition,hyperphosphorylation of Ikaros by CK2 leads to decreased Ikaros levels due to ubiquitinmediated degradation.Dephosphorylation of Ikaros by protein phosphatase 1 (PP1) acts in opposition to CK2 to increase Ikaros stability and restore Ikaros DNA binding ability and pericentromeric localization.Thus,the CK2 and PP1 pathways act in concert to regulate Ikaros activity in hematopoiesis and as a tumor suppressor.This highlights the importance of these signal transduction pathways as potential mediators of leukemogenesis via their role in regulating the activities of Ikaros.

  4. Functional analysis of Casein Kinase 1 in a minimal circadian system.

    Directory of Open Access Journals (Sweden)

    Gerben van Ooijen

    Full Text Available The Earth's rotation has driven the evolution of cellular circadian clocks to facilitate anticipation of the solar cycle. Some evidence for timekeeping mechanism conserved from early unicellular life through to modern organisms was recently identified, but the components of this oscillator are currently unknown. Although very few clock components appear to be shared across higher species, Casein Kinase 1 (CK1 is known to affect timekeeping across metazoans and fungi, but has not previously been implicated in the circadian clock in the plant kingdom. We now show that modulation of CK1 function lengthens circadian rhythms in Ostreococcustauri, a unicellular marine algal species at the base of the green lineage, separated from humans by ~1.5 billion years of evolution. CK1 contributes to timekeeping in a phase-dependent manner, indicating clock-mediated gating of CK1 activity. Label-free proteomic analyses upon overexpression as well as inhibition revealed CK1-responsive phosphorylation events on a set of target proteins, including highly conserved potentially clock-relevant cellular regulator proteins. These results have major implications for our understanding of cellular timekeeping and can inform future studies in any circadian organism.

  5. Comparative evaluation on fatty acid and Matricaria recutita essential oil incorporated into casein-based film.

    Science.gov (United States)

    Aliheidari, Nahal; Fazaeli, Mahboubeh; Ahmadi, Reza; Ghasemlou, Mehran; Emam-Djomeh, Zahra

    2013-05-01

    Sodium caseinate composite films containing lipids-oleic acid (OA), stearic acid (SA), or Matricaria recutita essential oil (MEO) - were prepared through emulsification and their physical, thermal, mechanical, and barrier properties were evaluated and compared. Furthermore, their antimicrobial effectiveness against Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli was studied. Emulsified films were softer, less rigid, and more stretchable than pure films. The films' water vapor barrier properties were found to decrease upon the addition of lipid content; this effect was greatly reduced when MEO was added. The presence of OA/SA and MEO decreased tensile strength and elastic modulus but increased the elongation at break. Thermal analysis of all emulsified films showed two endothermic peaks; these results confirmed those obtained by SEM studies, where a partial separation of the two phases occurred. The films' antimicrobial activities were increased by incorporating lipids, particularly those containing MEO, which were more effective against the studied bacteria. This work showed that when taking all the studied variables into account, films formulated with MEO were found most suitable for various food applications. PMID:23415659

  6. Soybean and casein hydrolysates induce grapevine immune responses and resistance against Plasmopara viticola.

    Science.gov (United States)

    Lachhab, Nihed; Sanzani, Simona M; Adrian, Marielle; Chiltz, Annick; Balacey, Suzanne; Boselli, Maurizio; Ippolito, Antonio; Poinssot, Benoit

    2014-01-01

    Plasmopara viticola, the causal agent of grapevine downy mildew, is one of the most devastating grape pathogen in Europe and North America. Although phytochemicals are used to control pathogen infections, the appearance of resistant strains and the concern for possible adverse effects on environment and human health are increasing the search for alternative strategies. In the present investigation, we successfully tested two protein hydrolysates from soybean (soy) and casein (cas) to trigger grapevine resistance against P. viticola. On Vitis vinifera cv. Marselan plants, the application of soy and cas reduced the infected leaf surface by 76 and 63%, as compared to the control, respectively. Since both hydrolysates might trigger the plant immunity, we investigated their ability to elicit grapevine defense responses. On grapevine cell suspensions, a different free cytosolic calcium signature was recorded for each hydrolysate, whereas a similar transient phosphorylation of two MAP kinases of 45 and 49 kDa was observed. These signaling events were followed by transcriptome reprogramming, including the up-regulation of defense genes encoding pathogenesis-related (PR) proteins and the stilbene synthase enzyme responsible for the biosynthesis of resveratrol, the main grapevine phytoalexin. Liquid chromatography analyses confirmed the production of resveratrol and its dimer metabolites, δ- and ε-viniferins. Overall, soy effects were more pronounced as compared to the cas ones. Both hydrolysates proved to act as elicitors to enhance grapevine immunity against pathogen attack. PMID:25566290

  7. Troglitazone inhibits endothelial cell proliferation through suppression of casein kinase 2 activity

    International Nuclear Information System (INIS)

    Troglitazone, an agonist of peroxisome proliferator activated receptorγ (PPARγ), has been reported to inhibit endothelial cell proliferation by suppressing Akt activation. Recently, it has been also proposed that phosphatase and tensin homolog deleted from chromosome 10 (PTEN) plays an important role in such effect of troglitazone. However, the mechanism of how troglitazone regulates PTEN remains to be elucidated. We therefore investigated the effects of troglitazone on casein kinase 2 (CK2), which is known to negatively regulate PTEN activity. Troglitazone significantly inhibited serum-induced proliferation of HUVEC in a concentration dependent manner. Serum-induced Akt and its downstream signaling pathway activation was attenuated by troglitazone (10 μM) pretreatment. The phosphorylation of PTEN, which was directly related to Akt activation, was decreased with troglitazone pretreatment and was inversely proportional to CK2 activity. DRB, a CK2 inhibitor, also showed effects similar to that of troglitazone on Akt and its downstream signaling molecules. In conclusion, our results suggest that troglitazone inhibits proliferation of HUVECs through suppression of CK2 activity rendering PTEN to remain activated, and this effect of troglitazone in HUVECs seems to be PPARγ independent

  8. Effect Of Five Proteases Including Alcalase, Flavourzyme, Papain, Proteinase K And Trypsin On Antioxidative Activities Of Casein Hydrolysate From Goat Milk

    Directory of Open Access Journals (Sweden)

    Shu Guowei

    2015-12-01

    Full Text Available Oxidation was related to the pathogenesis of human diseases. Adequate intake of antioxidant activity of food can reduce the levels of free radicals, prevent lipid peroxidation, and help the body against diseases. In the paper, casein from goat milk was hydrolyzed by five commercial proteases, namely, Alcalase, flavourzyme, papain, proteinase K and trypsin. The antioxidant activities of casein hydrolysates were assessed by evaluating hydrolysis degree, DPPH radical-scavenging activity, metal-chelating activity and superoxide radical scavenging activity. The results showed as follows: the DH of proteinase K, Alcalase, and trypsin were higher significantly than those of papain and flavourzyme. The Fe2+-chelation activity and superoxide radical scavenging activity of casein hydrolysates from goat milk by Alcalase was higher than the others, the DPPH scavenging activities of casein hydrolysates by Alcalase and papain was higher than the others and the DPPH scavenging activities by Alcalase and papain had no significant diffierence (p<0.05, so the optimal proteinase for hydrolysis casein from goat milk to produce antioxidant peptide was Alcalase.

  9. Polydimethylsiloxane composites containing 1,2-naphtoquinone 4-sulphonate as unique dispositive for estimation of casein in effluents from dairy industries.

    Science.gov (United States)

    Muñoz-Ortuño, M; Argente-García, A; Moliner-Martínez, Y; Molins-Legua, C; Campíns-Falcó, P

    2015-05-11

    A unique dispositive to determine casein which is the most abundant protein in dairy sewages has been proposed. In this sensing technology, the derivatization reagent 1,2-naphtoquininone 4-sulphonate (NQS) is embedded into a polydimethylsiloxane-tetraethylortosilicate-SiO2 nanoparticles composite (PDMS-TEOS-SiO2NPs). When the composite is immersed into the samples, casein is extracted from the solution and derivatized inside the PDMS matrix after 10 min at 100°C. The sensing support changes its color from yellow to orange depending on the casein concentration. Quantitative analysis can be carried out by measuring the absorbance with a reflection probe or by image-processing tool (GIMP). This sensor provides good sensitivity and precision (RSD% <12%). The method validation has been done by applying the biocinchoninic acid method (BCA). Moreover, semiquatitative analysis of casein can be performed by visual observation. Taking into account the advantages of small size, rapidity, simplicity, good stability and high compatibility in aqueous solution, this sensor is expected to have potential practical applications for in-situ determination of casein. Finally the method has been applied to analyze effluents from dairy industries. PMID:25911427

  10. Stimulation of casein kinase II by epidermal growth factor: Relationship between the physiological activity of the kinase and the phosphorylation state of its beta subunit

    International Nuclear Information System (INIS)

    To determine relationships between the hormonal activation of casein kinase II and its phosphorylation state, epidermal growth factor (EGF)-treated and EGF-naive human A-431 carcinoma cells were cultured in the presence of [32P]orthophosphate. Immunoprecipitation experiments indicated that casein kinase II in the cytosol of EGF-treated cells contained approximately 3-fold more incorporated [32P]phosphate than did its counterpart in untreated cells. Levels of kinase phosphorylation paralleled levels of kinase activity over a wide range of EGF concentrations as well as over a time course of hormone action. Approximately 97% of the incorporated [32P]phosphate was found in the β subunit of casein kinase II. Both activated and hormone-naive kinase contained radioactive phosphoserine and phosphothreonine but no phosphotyronsine. On the basis of proteolytic mapping experiments, EGF treatment of A-431 cells led to an increase in the average [32P]phosphate content (i.e., hyperphosphorylation) of casein kinase II β subunit peptides which were modified prior to hormone treatment. Finally, the effect of alkaline phosphatase on the reaction kinetics of activated casein kinase II indicated that hormonal stimulation of the kinase resulted from the increase in its phosphorylation state

  11. Protective effect of casein phosphopeptide-amorphous calcium phosphate on enamel erosion: Atomic force microscopy studies.

    Science.gov (United States)

    Ceci, Matteo; Mirando, Maria; Beltrami, Riccardo; Chiesa, Marco; Poggio, Claudio

    2015-01-01

    The aim of this study was to investigate the in vitro effect of a casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste (GC Tooth Mousse- TM, GC Corporation, Tokyo, Japan) on preventing enamel erosion, by using Atomic Force Microscopy (AFM). 30 human incisors, were equally assigned to 6 groups: intact enamel, enamel + soft drink, enamel + TM, enamel + TM + soft drink, enamel + soft drink + TM, enamel + soft drink + TM + soft drink. Specimens were observed through atomic force microscopy (AFM). The most common topographical parameters were determined, such as the surface roughness (Rrms ). The use of soft drink on intact enamel has roughened the surface of the sample. The application of the CPP-ACP paste on non-treated enamel made the surface smoother. A significant decrease in roughness was seen after remineralization with CPP-ACP paste. Significant differences were recorded when comparing softened enamel with softened enamel remineralized with CPP-ACP paste. Comparing eroded enamel with demineralized/remineralized specimens, the application of a CPP-ACP paste leads to a significant reduction in roughness values. AFM images of enamel surface treated with CPP-ACP resulted in less morphological changes of the tooth substrate when compared with the only eroded enamel surface morphology; thus, indicating that CPP-ACP paste promoted remineralization. Specimens' surface roughness remained similar regardless that the protective agent is used before or after exposure to coke or between two demineralizing cycles. The results confirmed the effectiveness of the CPP-ACP paste on preventing enamel erosion produced by soft drinks. PMID:25917931

  12. Synthesis of sodium caseinate-calcium carbonate microspheres and their mineralization to bone-like apatite

    Science.gov (United States)

    Xu, Zhewu; Liang, Guobin; Jin, Lin; Wang, Zhenling; Xing, Chao; Jiange, Qing; Zhang, Zhiguang

    2014-06-01

    Phosphoproteins can induce and stabilize calcium carbonate (CaCO3) vaterite, which has desirable features for high reactivity. The purpose of this study was to synthesize bioactive CaCO3 microspheres for bone regeneration. Sodium caseinate (NaCas)-containing CaCO3 microspheres, with the crystal phase of vaterite, were synthesized by fast precipitation in an aqueous solution of CaCl2, Na2CO3, and 2 mg/mL of NaCas. The uniform microspheres exhibited rougher surfaces and lower negative charges than CaCO3 particles without NaCas addition. Fourier-transform infrared spectroscopy (FT-IR) of the microspheres showed characteristic peaks or bands corresponding to phosphate and hydroxyl groups. Thermogravimetric analysis (TGA) curves exhibited approximately 5% weight loss below 600 °C due to the decomposition of NaCas. Scanning electron microscope (SEM) images showed lath-like hydroxyapatite (HAp) on the surface after soaking in simulated body fluid (SBF) at 37 °C for 5 and 10 days. Energy dispersive X-ray spectrometry (EDS) revealed that the agglomerates were composed of Ca, C, O, P, Na, and Mg elements, and the Ca/P ratios ranged from 1.53 to 1.56. X-ray diffraction (XRD) patterns exhibited peaks characteristic of hydroxyapatite. The results of this study demonstrated that the addition of NaCas induced the formation of vaterite microspheres which possesses an enhanced apatite formation after soaking in SBF at 37 °C for 5 and 10 days. These NaCas-CaCO3 microspheres may be a potential biomaterial for bone regeneration.

  13. Effects of Whey, Caseinate, or Milk Protein Ingestion on Muscle Protein Synthesis after Exercise.

    Science.gov (United States)

    Kanda, Atsushi; Nakayama, Kyosuke; Sanbongi, Chiaki; Nagata, Masashi; Ikegami, Shuji; Itoh, Hiroyuki

    2016-01-01

    Whey protein (WP) is characterized as a "fast" protein and caseinate (CA) as a "slow" protein according to their digestion and absorption rates. We hypothesized that co-ingestion of milk proteins (WP and CA) may be effective for prolonging the muscle protein synthesis response compared to either protein alone. We therefore compared the effect of ingesting milk protein (MP) to either WP or CA alone on muscle protein synthesis after exercise in rats. We also compared the effects of these milk-derived proteins to a control, soy protein (SP). Male Sprague-Dawley rats swam for two hours. Immediately after exercise, one of the following four solutions was administered: WP, CA, MP, or SP. Individual rats were euthanized at designated postprandial time points and triceps muscle samples collected for measurement of the protein fractional synthesis rate (FSR). FSR tended to increase in all groups post-ingestion, although the initial peaks of FSR occurred at different times (WP, peak time = 60 min, FSR = 7.76%/day; MP, peak time = 90 min, FSR = 8.34%/day; CA, peak time = 120 min, FSR = 7.85%/day). Milk-derived proteins caused significantly greater increases (p < 0.05) in FSR compared with SP at different times (WP, 60 min; MP, 90 and 120 min; CA, 120 min). Although statistical analysis could not be performed, the calculated the area under the curve (AUC) values for FSR following this trend were: MP, 534.61; CA, 498.22; WP, 473.46; and SP, 406.18. We conclude that ingestion of MP, CA or WP causes the initial peak time in muscle protein synthesis to occur at different times (WP, fast; MP, intermediate; CA, slow) and the dairy proteins have a superior effect on muscle protein synthesis after exercise compared with SP. PMID:27271661

  14. Goat milk allergenicity as a function of αs₁-casein genetic polymorphism.

    Science.gov (United States)

    Ballabio, C; Chessa, S; Rignanese, D; Gigliotti, C; Pagnacco, G; Terracciano, L; Fiocchi, A; Restani, P; Caroli, A M

    2011-02-01

    Cow milk allergy is the most frequent allergy in the first years of life. Milk from other mammalian species has been suggested as a possible nutritional alternative to cow milk, but in several cases, the clinical studies showed a high risk of cross-reactivity with cow milk. In the goat species, αS₁-casein (αS₁-CN), coded by the CSN1S1 gene, is characterized by extensive qualitative and quantitative polymorphisms. Some alleles are associated with null (i.e., CSN1S1 0(1)) or reduced (i.e., CSN1S1 F) expression of the specific protein. The aim of this work was to obtain new information on goat milk and to evaluate its suitability for allergic subjects, depending on the genetic variation at αs₁-CN. Individual milk samples from 25 goats with different CSN1S1 genotypes were analyzed by sodium dodecyl sulfate PAGE and immunoblotting, using monoclonal antibodies specific for bovine α-CN and sera from children allergic to cow milk. A lower reaction was observed to 2 goat milk samples characterized by the CSN1S1 0(1)0(1) and 0(1)F genotypes. Moreover, a fresh food skin prick test, carried out on 6 allergic children, showed the lack of positive reaction to the 0(1)0(1) milk sample and only one weak reactivity to the 0(1)F sample. The risk of cross-reactivity between cow and goat milk proteins suggests the need for caution before using goat milk for infant formulas. However, we hypothesize that it can be used successfully in the preparation of modified formulas for selected groups of allergic patients. The importance of taking the individual goat CN genetic variation into account in further experimental studies is evident from the results of the present work. PMID:21257068

  15. Toward the rational design of protein kinase casein kinase-2 inhibitors.

    Science.gov (United States)

    Sarno, Stefania; Moro, Stefano; Meggio, Flavio; Zagotto, Giuseppe; Dal Ben, Diego; Ghisellini, Paola; Battistutta, Roberto; Zanotti, Giuseppe; Pinna, Lorenzo A

    2002-01-01

    Casein kinase-2 (CK2) probably is the most pleiotropic member of the protein kinase family, with more than 200 substrates known to date. Unlike the great majority of protein kinases, which are tightly regulated enzymes, CK2 is endowed with high constitutive activity, a feature that is suspected to underlie its oncogenic potential and possible implication in viral infections. This makes CK2 an attractive target for anti-neoplastic and antiviral drugs. Here, we present an overview of our present knowledge about CK2 inhibitors, with special reference to the information drawn from two recently solved crystal structures of CK2alpha in complex with emodin and with 4,5,6,7-tetrabromo-2-azabenzimidazole (TBB), this latter being the most specific CK2 inhibitor known to date. A comparison with a series of anthraquinone and xanthenone derivatives highlights the crucial relevance of the hydroxyl group at position 3 for inhibition by emodin, and discloses the possibility of increasing the inhibitory potency by placing an electron withdrawing group at position 5. We also present mutational data corroborating the relevance of two hydrophobic residues unique to CK2, Val66 and Ile174, for the interactions with emodin and TBB, but not with the flavonoid inhibitors quercetin and fisetin. In particular, the CK2alpha mutant V66A displays 27- and 11-fold higher IC(50) values with emodin and TBB, respectively, as compared with the wild-type, while the IC(50) value with quercetin is unchanged. The data presented pave the road toward the rational design of more potent and selective inhibitors of CK2 and the generation of CK2 mutants refractory to inhibition, useful to probe the implication of CK2 in specific cellular functions. PMID:12191608

  16. Protective potential of casein phosphopeptide amorphous calcium phosphate containing paste on enamel surfaces

    Directory of Open Access Journals (Sweden)

    Padmini Somasundaram

    2013-01-01

    Full Text Available Background: Dental caries remains the most common dental disease facing mankind. Prevention of initiation and interruption in progression of early lesions are the desirable modes of caries management. There is a scope for agents, which may be used to enhance anti - caries activity. This need has redirected research to develop novel preventive agents that can act as an adjunct to fluoride or independent of it. Casein Phosphopeptide - Amorphous Calcium Phosphate (CPP-ACP is one such agent that has been proposed to have anti cariogenic properties. Aim: The purpose of this in vitro study was to evaluate the effect of paste containing CPP-ACP, MI Paste, on enamel remineralization. Materials and Methods: This study consisted of 30 samples embedded in orthodontic resin with either the buccal or lingual surface exposed. The samples were assigned to either a CPP-ACP containing paste; Fluoridated toothpaste; or a control group. The groups were then subjected to cycling in a demineralizing solution and a remineralizing solution. Groups II and III received prior application of MI paste and Fluoridated toothpaste respectively followed by cycling in a demineralizing solution and a remineralizing solution. Following 14 days of cycling, the samples were sectioned and examined using confocal microscopy. The lesion depth, were evaluated. Statistical Analysis: Image Proplus software was used to analyze the images. The values were statistically evaluated using one - way ANOVA and Scheffe′s Test. Results and Conclusion: Within the limitations of the study it was concluded that enamel surfaces treated with the CPP-ACP paste exhibited the least lesion depths followed by the enamel surfaces treated with the fluoridated tooth paste and control group respectively.

  17. Characterization of microcapsulated β-carotene formed by complex coacervation using casein and gum tragacanth.

    Science.gov (United States)

    Jain, Ashay; Thakur, Deepika; Ghoshal, Gargi; Katare, O P; Shivhare, U S

    2016-06-01

    Complex coacervation in casein/gum tragacanth (CAS/GT) mixtures was studied as a function of pH, initial protein to polysaccharide mixing ratio (Pr:Ps), total biopolymer concentration, core material load and ionic strength. This study is aimed at understanding how these parameters influence the coacervation kinetics, the coacervate yield, and entrapment efficiency. At a Pr:Ps=2:1, an optimum pH of complex coacervation was found 4.35, at which the intensity of electrostatic interaction was maximum. At these conditions, the phase separation occurred the fastest and the final coacervate yield and entrapment efficiency were the largest. Moreover, the developed β-carotene loaded microcapsules formulation was found to have particle size 159.71±2.16μm, coacervates yield 82.51±0.412%, entrapment efficiency 79.36±0.541%. Varying the Pr:Ps shifted the value of optimum pH. Electrostatic interaction and formation of coacervates was confirmed by Fourier Transform Infra Red (FTIR) spectra. Size and surface properties of coacervates were studied using Scanning Electron Microscopy (SEM). Entrapment of core material within the coacervates was confirmed by Confocal Laser Scanning Microscope (CLSM). The resultant formulation was evaluated for release study and antioxidant activity. Stability of encapsulated β-carotene was evaluated under three levels of temperature (5, 25 and 40°C) for 3 months. Encapsulation strongly increased the stability of micronutrients. Our results advocate potential of microcapsules as a novel carrier for the safeguard and sustained release of micronutrient. PMID:26851204

  18. Functional analysis of Trichoderma reesei CKIIα2, a catalytic subunit of casein kinase II.

    Science.gov (United States)

    Wang, Mingyu; Yang, Hui; Zhang, Meiling; Liu, Kuimei; Wang, Hanbin; Luo, Yi; Fang, Xu

    2015-07-01

    Trichoderma reesei is the most important industrial cellulase-producing filamentous fungus. Although its molecular physiology has been investigated, the signal transduction pathways are not fully understood. In particular, the role of casein kinase II (CKII) is not yet clear. In this work, we carried out functional investigations on a catalytic subunit of CKII, CKIIα2. Comparison of the phenotypic features of T. reesei parent and Δck2α2 strains showed significant changes following ck2α2 disruption. T. reesei Δck2α2 form significantly smaller mycelial pellets in glucose-containing liquid minimum media, have shorter and fewer branch hyphae, produce smaller amounts of chitinases, produce more spores, show more robust growth on glucose-containing agar plates, and consume glucose at a significantly higher rate. Suggestions can be made that CKIIα2 governs chitinase expression, and the disruption of ck2α2 results in lower levels of chitinase production, leading to a weaker cell wall disruption capability, further resulting in weaker hyphal branching, which eventually leads to smaller mycelial pellets in liquid media. Further conclusions can be made that CKIIα2 is involved in repression of sporulation and glucose metabolism, which is consistent with the proposal that CKIIα2 represses global metabolism. These observations make the deletion of ck2α2 a potentially beneficial genetic disruption for T. reesei during industrial applications, as smaller mycelial pellets, more spores and more robust glucose metabolism are all desired traits for industrial fermentation. This work reports novel unique functions of a CKII catalytic subunit and is also the first genetic and physiological investigation on CKII in T. reesei. PMID:25833183

  19. Casein-Coated Fe5C2 Nanoparticles with Superior r2 Relaxivity for Liver-Specific Magnetic Resonance Imaging.

    Science.gov (United States)

    Cowger, Taku A; Tang, Wei; Zhen, Zipeng; Hu, Kai; Rink, David E; Todd, Trever J; Wang, Geoffrey D; Zhang, Weizhong; Chen, Hongmin; Xie, Jin

    2015-01-01

    Iron oxide nanoparticles have been extensively used as T2 contrast agents for liver-specific magnetic resonance imaging (MRI). The applications, however, have been limited by their mediocre magnetism and r2 relaxivity. Recent studies show that Fe5C2 nanoparticles can be prepared by high temperature thermal decomposition. The resulting nanoparticles possess strong and air stable magnetism, suggesting their potential as a novel type of T2 contrast agent. To this end, we improve the synthetic and surface modification methods of Fe5C2 nanoparticles, and investigated the impact of size and coating on their performances for liver MRI. Specifically, we prepared 5, 14, and 22 nm Fe5C2 nanoparticles and engineered their surface by: 1) ligand addition with phospholipids, 2) ligand exchange with zwitterion-dopamine-sulfonate (ZDS), and 3) protein adsorption with casein. It was found that the size and surface coating have varied levels of impact on the particles' hydrodynamic size, viability, uptake by macrophages, and r2 relaxivity. Interestingly, while phospholipid- and ZDS-coated Fe5C2 nanoparticles showed comparable r2, the casein coating led to an r2 enhancement by more than 2 fold. In particular, casein coated 22 nm Fe5C2 nanoparticle show a striking r2 of 973 mM(-1)s(-1), which is one of the highest among all of the T2 contrast agents reported to date. Small animal studies confirmed the advantage of Fe5C2 nanoparticles over iron oxide nanoparticles in inducing hypointensities on T2-weighted MR images, and the particles caused little toxicity to the host. The improvements are important for transforming Fe5C2 nanoparticles into a new class of MRI contrast agents. The observations also shed light on protein-based surface modification as a means to modulate contrast ability of magnetic nanoparticles. PMID:26379788

  20. Relationship between genetic polymorphism of κ-casein and quantitative milk yield traits in cattle breeds and crossbreds in Serbia

    Directory of Open Access Journals (Sweden)

    Đedović Radica

    2015-01-01

    Full Text Available The quality and yield of milk, and therefore of dairy products as well, depends directly on their protein composition. Milk gene proteins represent structural locuses that are functionally associated with milk production. In cattle selection a special importance is attached to the variability of kappa casein (κ-CN as one of the four casein milk proteins. Genotypes of 21 Simmental breed cows, 20 crossbred cows obtained by crossing of Simmental and Red Holstein breed, and 25 cows of autochthonous Busha breed were determined for kappa casein by restriction fragment length polymorphism analysis (PCR- RFLP of amplified DNA. The κ-CN genotype frequencies for Simmental breed were: 42.8; 47.6 and 9.6 % for AA, and BB genotype, for crossbreds: 75.0; 25.0 and 0.0% and for Busha individuals: 41.7; 50.0 and 8.3 %, respectively. Frequencies of A and B alleles, assessed on the basis of genotypic frequencies were 0.667 and 0.333 for Simmental breed, 0.875 and 0.125 for crossbreds and 0.667 and 0.333 for autochthonous Busha cattle breed, respectively. For confirming the phenotypic variability of milk yield in standard lactation, yield of milk fat and the content of milk fat per κ-CN genotypes within breeds observed the fixed model which included the breed effect, lactation in order and effect of κ-CN genotype (AA, and BB was used. The κ-CN genotype statistically significantly (p ≤0.05 influenced milk yield and highly significantly (p ≤0.01 milk fat yield, while the content of milk fat was not statistically significantly (p>0.05 influenced in examined animals.

  1. Tertiary and quaternary structural differences between two genetic variants of bovine casein by small-angle X-ray scattering

    Energy Technology Data Exchange (ETDEWEB)

    Pessen, H.; Kumosinski, T.F.; Farrell, H.M. Jr.; Brumberger, H. (Dept. of Agriculture, Eastern Regional Research Center, Philadelphia, PA (USA))

    1991-01-01

    The casein complexes of bovine milk consist of four major protein fractions, alpha s1, alpha s2, beta, and kappa. Colloidal particles of casein (termed micelles) contain inorganic calcium and phosphate; they are very roughly spherical with an average radius of 650 A. Removal of Ca2+ leads to the formation of smaller protein aggregates with an average radius of 94 A. Two genetic variants, A and B, of the predominant fraction, alpha s1-casein, result in milks with markedly different physical properties, such as solubility and heat stability. To investigate the molecular basis for these differences, small-angle X-ray scattering was performed on the respective colloidal micelles and submicelles. Scattering curves for submicelles of both variants showed multiple Gaussian character; data for the B variant were previously interpreted in terms of two concentric regions of different electron density, i.e., a compact core and a relatively loose shell. For the submicelle of A, there was a third Gaussian, reflecting a negative contribution due to interparticle interference. Molecular parameters for submicelles of both A and B are in agreement with hydrodynamic data in the literature. Data for the micelles, for which scattering yields cross-sectional information, were fitted by a sum of three Gaussians for both variants; for these, the corresponding two lower radii of gyration represent the two concentric regions of the submicelles, while the third reflects the average packing of submicelles within the micellar cross section. Most of the molecular parameters obtained showed small but consistent differences between A and B, but for submicelles within the micelle several differences were particularly notable: A has a greater molecular weight for the compact region of the constituent submicelle (82,000 vs 60,000) and a much greater submicellar packing number.

  2. Cloning and expression in Escherichia coli of a new gene of Schistosoma japonicum encoding casein kinase Ⅱ beta subunit

    Institute of Scientific and Technical Information of China (English)

    彭寨玉; 余新炳; 吴忠道; 徐劲; 吴德; 李孜

    2004-01-01

    Background Nowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase Ⅱ beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E.coli).Methods The ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E.coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot. Results A 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase Ⅱ beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E.coli JM109.The recombinant protein could be recognized by the S. japonicum infected rabbit serum. Conclusion The full-length cDNA sequences encoding S. japonicum casein kinase Ⅱ beta subunit were firstly sequenced, cloned, and expressed in E.coli.

  3. Tertiary and quaternary structural differences between two genetic variants of bovine casein by small-angle X-ray scattering

    International Nuclear Information System (INIS)

    The casein complexes of bovine milk consist of four major protein fractions, alpha s1, alpha s2, beta, and kappa. Colloidal particles of casein (termed micelles) contain inorganic calcium and phosphate; they are very roughly spherical with an average radius of 650 A. Removal of Ca2+ leads to the formation of smaller protein aggregates with an average radius of 94 A. Two genetic variants, A and B, of the predominant fraction, alpha s1-casein, result in milks with markedly different physical properties, such as solubility and heat stability. To investigate the molecular basis for these differences, small-angle X-ray scattering was performed on the respective colloidal micelles and submicelles. Scattering curves for submicelles of both variants showed multiple Gaussian character; data for the B variant were previously interpreted in terms of two concentric regions of different electron density, i.e., a compact core and a relatively loose shell. For the submicelle of A, there was a third Gaussian, reflecting a negative contribution due to interparticle interference. Molecular parameters for submicelles of both A and B are in agreement with hydrodynamic data in the literature. Data for the micelles, for which scattering yields cross-sectional information, were fitted by a sum of three Gaussians for both variants; for these, the corresponding two lower radii of gyration represent the two concentric regions of the submicelles, while the third reflects the average packing of submicelles within the micellar cross section. Most of the molecular parameters obtained showed small but consistent differences between A and B, but for submicelles within the micelle several differences were particularly notable: A has a greater molecular weight for the compact region of the constituent submicelle (82,000 vs 60,000) and a much greater submicellar packing number

  4. Single application of casein phosphopeptide-amorphous calcium phosphate paste-based paste prevents in vitro erosive wear

    OpenAIRE

    Daniela Silva Barroso de Oliveira; Gabriela Cristina Santin; Heitor Marques Honorio; Daniela Rios; Patricia Hernandes Gaton; Léa Assed Bezerra da Silva; Fabio Lourenco Romano; Raquel Assed Bezerra da Silva

    2016-01-01

    Aim: The purpose of this study was to evaluate, in vitro, the effect of a single application of a casein phosphopeptide-amorphous calcium phosphate paste (CPP-ACP) in comparison with a fluoride varnish, in enamel of bovine teeth, after erosive challenge using cola soft drink. Materials and Methods: Forty-five enamel specimens were obtained from bovine teeth, selected according to the surface hardness and randomly divided into three groups: G1 - Fluoride varnish, G2 - CPP-ACP-based paste, and ...

  5. Comparing the Effects of Whey Extract and Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) on Enamel Microhardness

    OpenAIRE

    Rezvani, Mohammad Bagher; Karimi, Mehrdad; Akhavan Rasoolzade, Raheleh; Haghgoo, Roza

    2015-01-01

    Statement of the Problem With the recent focus of researches on the development of non-invasive treatment modalities, the non-invasive treatment of early carious lesions by remineralization would bring a major advance in the clinical management of these dental defects. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is considered to be effective in tooth remineralization. Purpose The aim of this in-vitro study was to compare the effects of whey and CPP-ACP in increasing the enamel...

  6. The effect of casein phosphopeptide amorphous calcium phosphate on the in vitro shear bond strength of orthodontic brackets

    OpenAIRE

    Park, Sun-Youn; Cha, Jung-Yul; Kim, Kyoung-Nam; Hwang, Chung-Ju

    2013-01-01

    Objective The purpose of this study was to evaluate the effect of casein phosphopeptide amorphous calcium phosphate (CPP-ACP) on the shear bond strength (SBS) of brackets bonded to non-demineralized teeth with either phosphoric acid etching or self-etching primer. Methods Sixty human premolars were randomly assigned to 1 of 4 treatment groups (n = 15 each): phosphoric acid etching (group 1); self-etching primer (group 2); CPP-ACP for 2 weeks + phosphoric acid etching (group 3), and CPP-ACP fo...

  7. Hydrolysis with Cucurbita ficifolia serine protease reduces antigenic response to bovine whey protein concentrate and αs-casein

    OpenAIRE

    Babij, Konrad; Bajzert, Joanna; Dąbrowska, Anna; Szołtysik, Marek; Zambrowicz, Aleksandra; Lubec, Gert; Stefaniak, Tadeusz; Willak-Janc, Ewa; Chrzanowska, Józefa

    2015-01-01

    In the present study the effect of hydrolysis with non-commercial Cucurbita ficifolia serine protease on a reduction of the IgE and IgG binding capacity of whey protein concentrate and αs-casein was investigated. The intensity of the protein degradation was analyzed by the degree of hydrolysis, the free amino groups content and RP-HPLC. The ability to bind the antibodies by native proteins and their hydrolysates was determined using a competitive ELISA test. Deep hydrolysis contributed to a s...

  8. Casein Kinase 2α Regulates Glioblastoma Brain Tumor Initiating Cell Growth through the β-Catenin Pathway

    OpenAIRE

    Nitta, Ryan T; Gholamin, Sharareh; Feroze, Abdullah H.; Agarwal, Maya; Cheshier, Samuel H.; Mitra, Siddhartha S.; Li, Gordon

    2014-01-01

    Glioblastoma (GBM) is the most common and fatal primary brain tumor in humans and it is essential that new and better therapies are developed to treat this disease. Previous research suggests that casein kinase 2 (CK2), may be a promising therapeutic target for GBMs. CK2 has enhanced expression or activity in numerous cancers, including GBM and it has been demonstrated that inhibitors of CK2 regressed tumor growth in GBM xenograft mouse models. Our studies demonstrate that the CK2 subunit, CK...

  9. Iron (II)-chelating activity of buffalo αS-casein hydrolysed by corolase PP, alcalase and flavourzyme

    OpenAIRE

    Jaiswal, Arvind; Bajaj, Rajesh; Mann, Bimlesh; Lata, Kiran

    2014-01-01

    Iron is a vital substance for human health which participates in many biochemical reactions. It also act as initiator for many harmful oxidative process. Buffalo αS-casein enriched fraction (80 %) was hydrolysed independently by corolase PP (H1), alcalase (H2), flavourzyme (H3) and sequentially by alcalase-flavourzyme (H4). After ultrafiltration (10 and 3 kDa) hydrolysates were analysed for their iron chelation activity using ferrozine. For H1 group of hydrolysates highest iron (II)-chelation...

  10. Effects of casein, whey and soy proteins on volumetric bone density and bone strength in immunocompromised piglets

    DEFF Research Database (Denmark)

    Budek, Alicja Zofia; Bjørnvad, Charlotte; Mølgaard, Christian;

    2007-01-01

    Summary:Background and aims: Bone-promoting effect of different proteins in early life, under immunocompromised conditions, is unknown. We investigated effects of milk- and plantderived proteins on bone development in immunocompromised piglets. Methods: Newborn, colostrum-deprived piglets were...... assigned to a formula based on either casein (n=11), whey (n=11) or soy (n=10) as the protein source (each 55 g/L), and equal amounts of fat, carbohydrates, calcium and phosphorus. Results & Conclusion: Despite efforts to sustain immuno-protection (sow serum and antibiotic injections), some piglets became...

  11. Purification of the Rous sarcoma virus src kinase by casein-agarose and tyrosine-agarose affinity chromatography.

    OpenAIRE

    Fukami, Y.; Lipmann, F

    1985-01-01

    A simple and effective purification method for the src kinase, the transforming gene product of Rous sarcoma virus, has been developed by using affinity chromatography on casein-agarose and tyrosine-agarose columns. NaDodSO4/polyacrylamide gel electrophoresis and silver staining analysis showed that the purified kinase preparation was composed of a predominant polypeptide of 60,000-Da. In most of the preparations, however, three minor proteins (54,000, 52,000, and 15,000 Da) were also detecte...

  12. The usefulness of casein-specific IgE and IgG4 antibodies in cow's milk allergic children

    OpenAIRE

    Ito Komei; Futamura Masaki; Movérare Robert; Tanaka Akira; Kawabe Tsutomu; Sakamoto Tatsuo; Borres Magnus P

    2012-01-01

    Abstract Background Cow's milk allergy is one of the most common food allergies among younger children. We investigated IgE antibodies to milk, and IgE and IgG4 antibodies to casein, α-lactalbumin and β-lactoglobulin in cow's milk allergic (CMA) and non-allergic (non-CMA) children in order to study their clinical usefulness. Methods Eighty-three children with suspected milk allergy (median age: 3.5 years, range: 0.8-15.8 years) were diagnosed as CMA (n = 61) or non-CMA (n = 22) based on an op...

  13. Selenized milk casein in the diet of BALB/c nude mice reduces growth of intramammary MCF-7 tumors

    International Nuclear Information System (INIS)

    Dietary selenium has the potential to reduce growth of mammary tumors. Increasing the Se content of cows’ milk proteins is a potentially effective means to increase Se intake in humans. We investigate the effects of selenized milk protein on human mammary tumor progression in immunodeficient BALB/c nude mice. Four isonitrogenous diets with selenium levels of 0.16, 0.51, 0.85 and 1.15 ppm were formulated by mixing low- and high-selenium milk casein isolates with a rodent premix. MCF-7 cells were inoculated into the mammary fat pad of female BALB/c nude mice implanted with slow-release 17 β-estradiol pellets. Mice with palpable tumors were randomly assigned to one of the four diets for 10 weeks, during which time weekly tumor caliper measurements were conducted. Individual growth curves were fit with the Gompertz equation. Apoptotic cells and Bcl-2, Bax, and Cyclin D1 protein levels in tumors were determined. There was a linear decrease in mean tumor volume at 70 days with increasing Se intake (P < 0.05), where final tumor volume decreased 35% between 0.16 and 1.15 ppm Se. There was a linear decrease in mean predicted tumor volume at 56, 63 and 70 days, and the number of tumors with a final volume above 500 mm3, with increasing Se intake (P < 0.05). This tumor volume effect was associated with a decrease in the proportion of tumors with a maximum growth rate above 0.03 day-1. The predicted maximum volume of tumors (Vmax) and the number of tumors with a large Vmax, were not affected by Se-casein. Final tumor mass, Bcl-2, Bax, and Cyclin D1 protein levels in tumors were not significantly affected by Se-casein. There was a significantly higher number of apoptotic cells in high-Se tumors as compared to low-Se tumors. Taken together, these results suggest that turnover of cells in the tumor, but not its nutrient supply, were affected by dairy Se. We have shown that 1.1 ppm dietary Se from selenized casein can effectively reduce tumor progression in an MCF-7 xenograft

  14. Short communication: Casein hydrolysate and whey proteins as excipients for cyanocobalamin to increase intestinal absorption in the lactating dairy cow.

    Science.gov (United States)

    Artegoitia, V M; de Veth, M J; Harte, F; Ouellet, D R; Girard, C L

    2015-11-01

    Bioavailability of vitamin B12 is low in humans and animals. Improving vitamin B12 absorption is important for optimal performance in dairy cows and for increasing vitamin B12 concentrations in milk for human consumption. However, when supplemented in the diet, 80% of synthetic vitamin B12, cyanocobalamin (CN-CBL), is degraded in the rumen of dairy cows and only 25% of the amount escaping destruction in the rumen disappears from the small intestine between the duodenal and ileal cannulas. In pigs, vitamin B12 from milk is more efficiently absorbed than synthetic CN-CBL. The objective of this study was to determine the efficacy of casein hydrolysate and whey proteins as excipients for CN-CBL to increase portal-drained viscera (PDV) flux of the vitamin in lactating dairy cows. Four multiparous lactating Holstein cows (237 ± 17 DIM) equipped with a rumen cannula and catheters in the portal vein and a mesenteric artery were used in a randomized Youden square design. They were fed every 2 h to maintain steady digesta flow. On experimental days, they received a postruminal bolus of (1) CN-CBL alone (0.1 g), (2) CN-CBL (0.1 g) + casein hydrolysate (10 g), or (3) CN-CBL (0.1 g) + whey proteins (10 g). Starting 30 min after the bolus, blood samples were taken simultaneously from the 2 catheters every 15 min during the first 2 h and then every 2 h until 24 h postbolus. Milk yield, DMI, and vitamin B12 portal-arterial difference and PDV flux were analyzed using the MIXED procedure of SAS. Milk yield and DMI were not affected by treatments. The portal-arterial difference of vitamin B12 during the 24-h period following the bolus of vitamin was greater when the vitamin was given in solution with casein hydrolysate (2.9 ± 4.6 pg/mL) than alone (-17.5 ± 5.2 pg/mL) or with whey protein (-13.4 ± 4.2 pg/mL). The treatment effects were similar for the PDV flux. The present results suggest that CN-CBL given with casein hydrolysate increases vitamin B12 absorption as compared with

  15. Whey Protein Delays Gastric Emptying and Suppresses Plasma Fatty Acids and Their Metabolites Compared to Casein, Gluten, and Fish Protein

    DEFF Research Database (Denmark)

    Stanstrup, Jan; Schou, Simon S; Holmer-Jensen, Jens;

    2014-01-01

    studies, the WI meal caused a decreased rate of gastric emptying compared to the other test meals. The WI meal also caused elevated levels of a number of amino acids, possibly stimulating insulin release leading to reduced plasma glucose. The WI meal also caused decreased levels of a number of fatty acids......Whey protein has been demonstrated to improve fasting lipid and insulin response in overweight and obese individuals. To establish new hypotheses for this effect and to investigate the impact of stomach emptying, we compared plasma profiles after intake of whey isolate (WI), casein, gluten (GLU...

  16. YY1 represses beta-casein gene expression by preventing the formation of a lactation-associated complex.

    OpenAIRE

    Raught, B; Khursheed, B; Kazansky, A; Rosen, J.

    1994-01-01

    Site-specific mutagenesis of the highly conserved milk box (-140 to -110) region suggested that beta-casein expression is regulated by a hormone-mediated relief of repression (M. Schmitt-Ney, W. Doppler, R. K. Ball, and B. Groner, Mol. Cell. Biol. 11:3745-3755, 1991). However, when this sequence was placed upstream of a heterologous thymidine kinase promoter, it activated reporter gene expression. This apparent paradox was resolved when the trans-acting factor YY1, capable of acting as both a...

  17. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells.

    OpenAIRE

    M. L. Li; Aggeler, J; Farson, D A; Hatier, C; Hassell, J; Bissell, M.J.

    1987-01-01

    When primary mouse mammary epithelial cells are cultured on plastic, they rapidly lose their ability to synthesize and secrete most milk proteins even in the presence of lactogenic hormones, whereas cells cultured on released type I collagen gels show greatly enhanced mRNA levels and secretion rates of beta-casein and of some other milk proteins. We show here that culture on a reconstituted basement membrane from Engelbreth-Holm-Swarm tumor (EHS) allows greater than 90% of cells to produce hi...

  18. A peptidomic approach to study the contribution of added casein proteins to the peptide profile in Spanish dry-fermented sausages.

    Science.gov (United States)

    Mora, Leticia; Escudero, Elizabeth; Aristoy, M-Concepción; Toldrá, Fidel

    2015-11-01

    Peptidomics is a necessary alternative in the analysis of naturally generated peptides in dry-fermented processing. The intense proteolysis occurred during the processing of dry-fermented sausages is due to the action of endopeptidases and exopeptidases from both, endogenous muscle origin and lactic acid bacteria (LAB) added in the starter. Sodium caseinate is frequently used as an additive in this type of products because of its emulsifying properties, and consequently influences the protein profile available during the proteolysis. In this study, a mass spectrometry approach has been used to determine the impact of added sodium caseinate in the final peptide profile as well as to analyse its possible influence in the presence of certain previously described casein-derived bioactive peptides. PMID:26116420

  19. Homo-fermentative production of D-lactic acid by Lactobacillus sp. employing casein whey permeate as a raw feed-stock.

    Science.gov (United States)

    Prasad, Saurav; Srikanth, Katla; Limaye, Anil M; Sivaprakasam, Senthilkumar

    2014-06-01

    Casein whey permeate (CWP), a lactose-enriched dairy waste effluent, is a viable feed stock for the production of value-added products. Two lactic acid bacteria were cultivated in a synthetic casein whey permeate medium with or without pH control. Lactobacillus lactis ATCC 4797 produced D-lactic acid (DLA) at 12.5 g l(-1) in a bioreactor. The values of Leudking-Piret model parameters suggested that lactate was a growth-associated product. Batch fermentation was also performed employing CWP (35 g lactose l(-1)) with casein hydrolysate as a nitrogen supplement in a bioreactor. After 40 h, L. lactis produced 24.3 g lactic acid l(-1) with an optical purity >98 %. Thus CWP may be regarded as a potential feed-stock for DLA production. PMID:24563313

  20. In Vitro Proliferation and Anti-Apoptosis of the Papain-Generated Casein and Soy Protein Hydrolysates towards Osteoblastic Cells (hFOB1.19

    Directory of Open Access Journals (Sweden)

    Xiao-Wen Pan

    2015-06-01

    Full Text Available Casein and soy protein were digested by papain to three degrees of hydrolysis (DH 7.3%–13.3%, to obtain respective six casein and soy protein hydrolysates, aiming to clarify their in vitro proliferation and anti-apoptosis towards a human osteoblastic cell line (hFOB1.19 cells. Six casein and soy protein hydrolysates at five levels (0.01–0.2 mg/mL mostly showed proliferation as positive 17β-estradiol did, because they conferred the osteoblasts with cell viability of 100%–114% and 104%–123%, respectively. The hydrolysates of higher DH values had stronger proliferation. Casein and soy protein hydrolysates of the highest DH values altered cell cycle progression, and enhanced cell proportion of S-phase from 50.5% to 56.5% and 60.5%. The two also antagonized etoposide- and NaF-induced osteoblast apoptosis. In apoptotic prevention, apoptotic cells were decreased from 31.6% to 22.6% and 15.6% (etoposide treatment, or from 19.5% to 17.7% and 12.4% (NaF treatment, respectively. In apoptotic reversal, soy protein hydrolysate decreased apoptotic cells from 13.3% to 11.7% (etoposide treatment, or from 14.5% to 11.0% (NaF treatment, but casein hydrolysate showed no reversal effect. It is concluded that the hydrolysates of two kinds had estradiol-like action on the osteoblasts, and soy protein hydrolysates had stronger proliferation and anti-apoptosis on the osteoblasts than casein hydrolysates.

  1. Study on genetic variability at αS1 and αS2 casein loci in autochthonous goat populations of the Lombardy Alps

    OpenAIRE

    P. Crepaldi; D. Milani; P. Ramelli; M. Marilli; D. Meggiolaro

    2011-01-01

    In cattle, sheep and goats (Hayes et al., 1993) the casein genes form a cluster, on the chromosome 6, of four tightly linked genes, whose sequence is: αS1-β-αS2-k (Threadgill and Womack, 1990; Ferretti et al., 1990; Gallagher et al., 1994; Rijnkles et al., 1997). For this reason the alleles at these loci are generally inherited all together (Di Stasio and Mariani, 2000), with low recombination frequency. Thus it is worth noting the importance of describing the casein haplotyp...

  2. ANALYSIS OF POLYMORPHISM OF BETA CASEIN OF SLOVAK PINZGAU CATTLE BY PCR-RFLP FOR ALLELS A1 AND A2

    OpenAIRE

    MARTINA MILUCHOVÁ; ANNA TRAKOVICKÁ; M. GÁBOR

    2013-01-01

    The work was oriented to identification of -casein gene polymorphism and analysis of genotype structure in population of Slovak Pinzgau cattle. The material involved 89 cattle. Bovine genomic DNA was isolated by fenol-chlorophorm deprotenization and ethanol precipitation and used in order to estimate  - casein genotypes by means of PCR-RFLP method. The PCR products were digested with DdeI restriction enzyme. In the population included in the study there were homozygote genotype A1A1 (27 ani...

  3. A specific pattern of splicing for the horse $\\alpha$S1-Casein mRNA and partial genomic characterization of the relevant locus

    OpenAIRE

    Milenkovic, Dragan; Martin, Patrice; Guérin, Gérard; Leroux, Christine

    2002-01-01

    Mares' milk has a composition very different from that of cows' milk. It is much more similar to human milk, in particular in its casein fraction. This study reports on the sequence of a 994 bp amplified fragment corresponding to a horse $\\alpha$S1-Casein ($\\alpha$ S1-Cn) cDNA and its comparison with its caprine, pig, rabbit and human counterparts. The alignment of these sequences revealed a specific pattern of splicing for this horse primary transcript. As in humans, exons 3$'$, 6$'$ and 13$...

  4. Precalving and early lactation factors that predict milk casein and fertility in the transition dairy cow.

    Science.gov (United States)

    Rodney, Rachael M; Hall, Jenianne K; Westwood, Charlotte T; Celi, Pietro; Lean, Ian J

    2016-09-01

    Multiparous Holstein cows (n=82) of either high or low genetic merit (GM) (for milk fat + protein yield) were allocated to 1 of 2 diets in a 2×2 factorial design. Diets differed in the ratio of rumen-undegradable protein (RUP) to rumen-degradable protein (37% RUP vs. 15% RUP) and were fed from 21 d precalving to 150 days in milk. This study evaluated the effects of these diets and GM on concentrations of milk casein (CN) variants and aimed to identify precalving and early lactation variables that predict milk CN and protein yield and composition and fertility of dairy cows. It explored the hypothesis that low milk protein content is associated with lower fertility and extended this hypothesis to also evaluate the association of CN contents with fertility. Yields (kg/d) for CN variants were 0.49 and 0.45 of α-CN, 0.38 and 0.34 of β-CN, 0.07 and 0.06 for κ-CN, and 0.10 and 0.09 of γ-CN for high- and low-RUP diets, respectively. Increased RUP increased milk, CN, and milk protein yields. Increased GM increased milk protein and γ-CN yields and tended to increase milk CN yield. The effects of indicator variables on CN variant yields and concentrations were largely consistent, with higher body weight and α-amino nitrogen resulting in higher yields, but lower concentrations. An increase in cholesterol was associated with decreased CN variant concentrations, and disease lowered CN variant yield. A diet high in RUP increased proportion of first services that resulted in pregnancy from 41 to 58%. Increased precalving metabolizable protein (MP) balance decreased the proportion of first services that resulted in pregnancy when evaluated in a model containing CN percentage, milk protein yield, diet, and GM. This finding suggests that the positive effects of increasing dietary RUP on fertility may be curvilinear because cows with a very positive MP balance before calving were less fertile than those with a lower, but positive, MP balance. Prepartum MP balance was important

  5. Calcium bioaccessibility and uptake by human intestinal like cells following in vitro digestion of casein phosphopeptide-calcium aggregates.

    Science.gov (United States)

    Perego, Silvia; Del Favero, Elena; De Luca, Paola; Dal Piaz, Fabrizio; Fiorilli, Amelia; Cantu', Laura; Ferraretto, Anita

    2015-06-01

    Casein phosphopeptides (CPPs), derived by casein proteolysis, can bind calcium ions and keep them in solution. In vitro studies have demonstrated CPP-induced cell calcium uptake, depending on the formation of (CPP + calcium) complexes and on the degree of differentiation of the intestinal cells. With the present study, we address the persistence of the complexes and of the CPP-induced calcium uptake in intestinal like cells after the digestion process, thus examining their eligibility to serve as nutraceuticals. A calcium-preloaded CPP preparation of commercial origin (Ca-CPPs) was subjected to in vitro digestion. The evolution of the supramolecular structure of the Ca-CPP complexes was studied using laser-light and X-ray scattering. The bioactivity of the pre- and post-digestion Ca-CPPs was determined in differentiated Caco2 and HT-29 cells by video imaging experiments using Fura-2. We found that Ca-CPP aggregates keep a complex supramolecular organization upon digestion, despite getting smaller in size and increasing internal calcium dispersion. Concomitantly and most interestingly, digested Ca-CPPs clearly enhance the uptake of calcium ions, especially in Caco2 cells. In contrast, digestion depletes the ability of post-loaded decalcified-CPPs (Ca-dekCPPs), with a weaker internal structure, to induce calcium uptake. The enhanced bioactivity reached upon digestion strongly suggests a recognized role of Ca-CPPs, in the form used here, as nutraceuticals. PMID:25927875

  6. The investigation of the secondary structures of various peptide sequences of β-casein by the multicanonical simulation method

    Science.gov (United States)

    Yaşar, F.; Çelik, S.; Köksel, H.

    2006-05-01

    The structural properties of Arginine-Glutamic acid-Leucine-Glutamic acid-Glutamic acid-Leucine-Asparagine-Valine-Proline-Glycine (RELEELNVPG, in one letter code), Glutamic acid-Glutamic acid-Glutamine-Glutamine-Glutamine-Threonine-Glutamic acid (EEQQQTE) and Glutamic acid-Aspartic acid-Glutamic acid-Leucine-Glutamine-Aspartic acid-Lysine-Isoleucine (EDELQDKI) peptide sequences of β-casein were studied by three-dimensional molecular modeling. In this work, the three-dimensional conformations of each peptide from their primary sequences were obtained by multicanonical simulations. With using major advantage of this simulation technique, Ramachandran plots were prepared and analysed to predict the relative occurrence probabilities of β-turn, γ-turn and helical structures. Structural predictions of these sequences of β-casein molecule indicate the presence of high level of helical structures and βIII-turns. The occurrence probabilities of inverse and classical β-turns were low. The probability of helical structure of each sequence significantly decreased when the temperature increased. Our results show these peptides have highly helical structure and better agreement with the results of spectroscopic techniques and other prediction methods.

  7. Hydrolysis with Cucurbita ficifolia serine protease reduces antigenic response to bovine whey protein concentrate and αs-casein.

    Science.gov (United States)

    Babij, Konrad; Bajzert, Joanna; Dąbrowska, Anna; Szołtysik, Marek; Zambrowicz, Aleksandra; Lubec, Gert; Stefaniak, Tadeusz; Willak-Janc, Ewa; Chrzanowska, Józefa

    2015-11-01

    In the present study the effect of hydrolysis with non-commercial Cucurbita ficifolia serine protease on a reduction of the IgE and IgG binding capacity of whey protein concentrate and αs-casein was investigated. The intensity of the protein degradation was analyzed by the degree of hydrolysis, the free amino groups content and RP-HPLC. The ability to bind the antibodies by native proteins and their hydrolysates was determined using a competitive ELISA test. Deep hydrolysis contributed to a significant reduction of immunoreactive epitopes present in WPC. In the case of IgE and IgG present in the serum pool of children with CMA, the lowest binding capacity was detected in the 24 h WPC hydrolysate, where the inhibition of the reaction with native WPC was ≤23 and ≤60 %, respectively. The analysis of the IgG reactivity in the antiserum of the immunized goat showed that the lowest antibody binding capacity was exhibited also by 24 h WPC hydrolysate at a concentration of 1000 μg/ml where the inhibition of the reaction with nWPC was ≤47 %. One-hour hydrolysis of α-casein was sufficient to significant reduction of the protein antigenicity, while the longer time (5 h) of hydrolysis probably lead to the appearance of new epitopes reactive with polyclonal. PMID:26036686

  8. The Impact of Kappa Casein Gene Polymorphism on Milk Components and Other Productive Performance Traits of Chinese Holstein Cattle

    Directory of Open Access Journals (Sweden)

    A. E. Hamza1, 2, Z. P.Yang1*, X. L. Wang1, R. J. Chen1, H. T. Wu1 and A. I. Ibrahim3

    2011-04-01

    Full Text Available Polymorphism of kappa casein gene (κ-Cn at exon 5 in 259 Chinese Holstein cattle was investigated using polymerase chain reaction-single strand conformation (PCR-SSCP technique, to verify its effect on milk composition (fat, protein, lactose, total solids (TS and solids not fat (SNF content and influence of other factors such as parity and season of calving; aiming at utilizing it as a genetic aid in selection to improve the quality of production of this herd. A 218 bp fragment containing exon 1V of kappa casein gene was amplified by PCR and SSCP was applied to identify the structural gene polymorphism of κ-Cn. General linear model (GLM was used to analyze differences between genotypes. The results indicated that κ-Cn genotypes significantly (P0.05 on TS and SNF. However, cows with genotype TT showed higher fat and protein contents (3.89 and 3.50%, respectively than those of genotypes CC and TC (3.62 and 3.66% for fat; 3.35 and 3.36 % for protein, respectively. Except for lactose content; parity had no effect on milk components. Also, the results disclosed that season of calving did not affect (P>0.05 milk composition. This study indicated that the κ-Cn genetic variants may be used as a genetic aid through increasing the frequency of desired genotypes to improve the quality of production of this herd.

  9. Casein kinase 1 controls the activation threshold of an α-arrestin by multisite phosphorylation of the interdomain hinge.

    Science.gov (United States)

    Herrador, Antonio; Livas, Daniela; Soletto, Lucía; Becuwe, Michel; Léon, Sébastien; Vincent, Olivier

    2015-06-01

    α-Arrestins play a key role as trafficking adaptors in both yeast and mammals. The yeast Rim8/Art9 α-arrestin mediates the recruitment of endosomal sorting complex required for transport (ESCRT) to the seven-transmembrane protein Rim21 in the ambient pH signaling RIM pathway. ESCRT is believed to function as a signaling platform that enables the proteolytic activation of the Rim101 transcription factor upon external alkalization. Here we provide evidence that the pH signal promotes the stable association of Rim8 with Rim21 at the plasma membrane. We show that Rim8 is phosphorylated in a pH-independent but Rim21-dependent manner by the plasma membrane-associated casein kinase 1 (CK1). We further show that this process involves a cascade of phosphorylation events within the hinge region connecting the arrestin domains. Strikingly, loss of casein kinase 1 activity causes constitutive activation of the RIM pathway, and, accordingly, pH signaling is activated in a phosphodeficient Rim8 mutant and impaired in the corresponding phosphomimetic mutant. Our results indicate that Rim8 phosphorylation prevents its accumulation at the plasma membrane at acidic pH and thereby inhibits RIM signaling. These findings support a model in which CK1-mediated phosphorylation of Rim8 contributes to setting a signaling threshold required to inhibit the RIM pathway at acidic pH. PMID:25851600

  10. Association of polymorphism κ-casein gene with longevity and lifetime production of Holstein-Friesian cows in Vojvodina

    Directory of Open Access Journals (Sweden)

    Dragomir Lukač

    2015-11-01

    Full Text Available The aim of this study was to identify frequencies of alleles and genotypes, to evaluate their frequency in population of Holstein-Friesian cows, and to investigate association of κ-casein genotypes with longevity and lifetime production of cows. Blood samples were collected from 192 Holstein cows which have finished their production. Cows were reared at one farm in Vojvodina in Serbia. In the current study the following ratios of κ-casein genotypes were found: 0.50 were AA, 0.40 were AB and 0.10 were BB. Frequencies of alleles A and B were 0.70 and 0.30, respectively. Based on these results, it can be concluded that cows with heterozygous genotype AB had the longest life (2809 days with the highest number of productive days (2062, while the cows which were homozygous recessive of genotype BB had the shortest life expectancy (2627 days and the lowest number productive days (1878. The same trend was observed with regard to the production results of cows of these genotypes, where statistically significant differences have been observed (P0.05. The effect of dominant genes in observed traits was greater than the additive effect of genes. From the ratio of the additive and dominant gene effects, it can be seen that the average degree of dominance in observed traits was positive.

  11. Competitive adsorption between beta-casein or beta-lactoglobulin and model milk membrane lipids at oil-water interfaces.

    Science.gov (United States)

    Waninge, Rianne; Walstra, Pieter; Bastiaans, Jan; Nieuwenhuijse, Hans; Nylander, Tommy; Paulsson, Marie; Bergenståhl, Björn

    2005-02-01

    This study investigated the competitive adsorption between milk proteins and model milk membrane lipids at the oil-water interface and its dependence on the state of the lipid dispersion and the formation of emulsions. Both protein and membrane lipid surface load were determined using a serum depletion technique. The membrane lipid mixture used was a model milk membrane lipid system, containing dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, milk sphingomyelin, dioleoylphosphatidylserine, and soybean phosphatidylinositol. The model composition mimics the lipid composition of natural milk fat globule membranes. The interactions were studied for two proteins, beta-lactoglobulin and beta-casein. The mixing order was varied to allow for differentiation between equilibrium structures and nonequilibrium structures. The results showed more than monolayer adsorption for most combinations. Proteins dominated at the oil-water interface in the protein-emulsified emulsion even after 48 h of exposure to a vesicular dispersion of membrane lipids. The membrane lipids dominated the oil-water interface in the case of the membrane lipid emulsified emulsion even after equilibration with a protein solution. Protein displacement with time was observed only for emulsions in which both membrane lipids and beta-casein were included during the emulsification. This study shows that kinetics controls the structures rather than the thermodynamic equilibrium, possibly resulting in structures more complex than an adsorbed monolayer. Thus, it can be expected that procedures such as the mixing order during emulsion preparation are of crucial importance to the emulsification performance. PMID:15686425

  12. Genetic polymorphism of beta-casein gene and its associations with milk traits in Holstein-Friesian cows

    Directory of Open Access Journals (Sweden)

    Teodor Bugeac

    2015-05-01

    Full Text Available In animal breeding finding and using effective genetic markers for improving important traits it is a continuous challenge. In this respect, several genetic markers were associated in cattle with increased milk production or a better milk quality. This proved to be a useful tool for improving certain traits by selecting individuals carriers of allelic variants that have an effect on a desirable trait. In particular, positive associations between certain alleles found at the milk protein loci with some milk production traits convincingly demonstrated in several cattle breeds. Although, in some cases the results obtained in various studies were not in agreement and varied between breeds or populations. Therefore the objective of this study was to establish associations (if any between alleles found at the beta-casein (CSN2 locus and some milk production traits (milk yield and fat, protein, casein and lactose content in a Holstein-Friesian population reared in Romania. Genetic variants at CSN2 locus were identified by isoelectric focusing (IEF of milk samples. In order to determine milk composition the samples were analysed with MilkoScan FT 6000. For the statistical analysis of data SPSS v.19 for Windows was used. At the CSN2 locus four alleles and seven genotypes were identified in the analyzed cattle population. The cows carriers of CSN2 A2 allele produced the highest milk yield and the highest milk protein content, this result being in agreement with other previous studies.

  13. Setting up of a liquid chromatography-high resolution tandem mass spectrometry method for the detection of caseins in food. A comparison with ELISA method

    Directory of Open Access Journals (Sweden)

    Daniela Gastaldi

    2013-06-01

    Full Text Available Determination of caseins in food matrices is usually performed by using the competitiveenzyme- linked immunosorbent assay (ELISA technique. However such a technique suffers from a number of limitations. Among these, the applicability to a narrow concentration range, a non linear (logarithmic response, a non-negligible cross-reactivity and a high cost per kit. At the time of the completion of this study, in case of ELISA positive feedback, there was poor availability in the literature of finding reliable instrumental methods able to determine both qualitatively and quantitatively this class of substances. In the present study, a liquid chromatography-high resolution tandem mass spectrometry (HPLC-HRMS/MS instrumental method was developed with a high resolution mass spectrometer (Orbitrap. Real samples of sausages in which caseins were detected by ELISA technique were analysed. A casein-free sample of ham was used as a blank. The analytical characteristics of the instrumental method were compared with those of a commercial ELISA test, declared specific for α- and β-casein.

  14. An evaluation of casein hydrolyzate in combination with antibiotic for bacterial cure and subsequent increase in milk yield in dairy cows

    Directory of Open Access Journals (Sweden)

    Silanikove Nissim

    2011-01-01

    Full Text Available Abstract Background A 3-yr study examined whether prepartum treatment with casein hydrolyzate in combination with antibiotic, as routinely used in Israel for dry cow therapy, improved bacterial cure and increased milk yield in subsequent lactations in comparison with treatment with antibiotic alone. The vast majority of bacterial isolates in samples collected prior to drying-off comprised coagulase-negative staphylococci, mostly as Staph. chromogenes. Results Bacterial cure associated with the combined treatment was 73.8% in cows, significantly higher than the 51.7% cure recorded when cows were treated only with antibiotic. During the study, the annual milk yield of non-casein hydrolyzate treated and treated control cows increased at ~2% per year, which is consistent with the national annual increase attributed to genetic selection. In cows treated with casein hydrolyzate the increase was 9% (above the 2% expected in the first lactation after the treatment, and 6.3% (above the 4% expected for 2 years in the second lactation after treatment. These increases were significantly higher than those in the controls and those expected through genetic improvement. Conclusions Treatment with casein hydrolyzate at dry-off was shown to be a viable mean to eliminate existing environmental bacterial infection, and to improve milk yield in the next lactation.

  15. Application of humidity-controlled dynamic mechanical analysis (DMA-RH) to moisture-sensitive edible casein films for use in food packaging

    Science.gov (United States)

    Protein-based and other hydrophilic thin films are promising materials for the manufacture of edible food packaging and other food and non-food applications. Calcium caseinate (CaCas) films are highly hygroscopic and physical characterization under broad environmental conditions is critical to appli...

  16. Rapid quantification of casein in skim milk using Fourier transform infrared spectroscopy, enzymatic perturbation, and multiway partial least squares regression: Monitoring chymosin at work.

    Science.gov (United States)

    Baum, A; Hansen, P W; Nørgaard, L; Sørensen, John; Mikkelsen, J D

    2016-08-01

    In this study, we introduce enzymatic perturbation combined with Fourier transform infrared (FTIR) spectroscopy as a concept for quantifying casein in subcritical heated skim milk using chemometric multiway analysis. Chymosin is a protease that cleaves specifically caseins. As a result of hydrolysis, all casein proteins clot to form a creamy precipitate, and whey proteins remain in the supernatant. We monitored the cheese-clotting reaction in real time using FTIR and analyzed the resulting evolution profiles to establish calibration models using parallel factor analysis and multiway partial least squares regression. Because we observed casein-specific kinetic changes, the retrieved models were independent of the chemical background matrix and were therefore robust against possible covariance effects. We tested the robustness of the models by spiking the milk solutions with whey, calcium, and cream. This method can be used at different stages in the dairy production chain to ensure the quality of the delivered milk. In particular, the cheese-making industry can benefit from such methods to optimize production control. PMID:27265175

  17. Lipopolysaccharide derived from the digestive tract activates inflammatory gene expression and inhibits casein synthesis in the mammary glands of lactating dairy cows.

    Science.gov (United States)

    Zhang, Kai; Chang, Guangjun; Xu, Tianle; Xu, Lei; Guo, Junfei; Jin, Di; Shen, Xiangzhen

    2016-03-01

    To meet the nutrition requirements of lactation, dairy cows are usually fed a high concentrate diet (HC). However, high-grain feeding causes subacute ruminal acidosis (SARA), a metabolic disorder that causes milk protein depression. This study aimed to investigate the effect of lipopolysaccharide (LPS) released in the rumen on inflammatory gene expression and casein synthesis in mammary glands of lactating dairy cows fed a HC diet. We found that milk protein was significantly decreased in the HC group after 15 weeks of feeding. Overall, LPS concentrations in the rumen fluid, lacteal artery and vein were increased in the HC group. Transcriptome microarray was used to evaluate alterations in the signaling pathway in mammary glands. Signaling pathways involved in inflammatory responses were activated, whereas those involved in protein synthesis were inhibited in the HC group. mRNA expression involved in inflammatory responses, including that of TLR4, NF-кB and pro-inflammatory genes, was increased in the HC group, while αs1-casein (CSN1S1), β-casein (CSN2), mTOR and S6K gene expression were decreased. Moreover, protein expression was consistent with the corresponding gene expression. After feeding with an HC diet, LPS derived from the rumen increased inflammatory gene expression and inhibited casein synthesis in the mammary glands of lactating dairy cows fed a HC diet. PMID:26893357

  18. Influence of human diets containing casein, soy protein isolate, and soy protein concentrate on serum cholesterol and lipoproteins in humans, rabbits and rats

    NARCIS (Netherlands)

    Raaij, van J.M.A.

    1982-01-01

    It is well known that feeding animals such as rabbits with semipurified diets containing animal proteins, as for example casein, results in hypercholesterolemia and atherosclerosis. On the other hand, diets containing vegetable proteins such as soybean protein maintain low levels of serum cholestero

  19. The effect of semipurified diets containing either casein or soybean protein on the concentration of serum cholesterol and the lipoprotein composition in rabbits

    NARCIS (Netherlands)

    Terpstra, A.H.M.

    1981-01-01

    This thesis deals with the effect of dietary casein and soybean protein on the concentration of serum cholesterol and the lipoprotein composition in rabbits. Special attention has been paid to the time course of the changes produced by the protein in the diet.After a short introduction, a review of

  20. Characterization of enzymatically induced aggregation of casein micelles in natural concentration by in situ static light scattering and ultra low shear viscosimetry

    DEFF Research Database (Denmark)

    Lehner, D.; Worning, Peder; G, Fritz; L, Ogendal; R, Bauer; Glatter, O.

    1999-01-01

    The aggregation of casein micelles in undiluted skim milk after the addition of chymosin was studied bystatic light scattering and ultra low shear viscometry. The static light scattering measurements were madewith two different sample thicknesses, 72 and 16 mum. The scattering data were analyzed ...