Hostname: page-component-78c5997874-94fs2 Total loading time: 0 Render date: 2024-11-08T20:28:02.237Z Has data issue: false hasContentIssue false
  • English
  • Français

Process optimisation for preparation of caseinophosphopeptides from Buffalo milk casein and their characterisation

Published online by Cambridge University Press:  23 July 2014

Prerna Saini ,
Bimlesh Mann ,
Rajesh Kumar ,
Rajan Sharma ,
Ram Ran Bijoy Singh  and
Alok Chatterjee
Prerna Saini
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132001, India
Bimlesh Mann*
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132001, India
Rajesh Kumar
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132001, India
Rajan Sharma
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132001, India
Ram Ran Bijoy Singh
Affiliation:
Division of Dairy Technology, National Dairy Research Institute, Karnal-132001, India
Alok Chatterjee
Affiliation:
Division of Dairy Technology, National Dairy Research Institute, Karnal-132001, India
*
*For correspondence; e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Caseinophosphopeptides (CPPs) are multifunctional bioactive peptides containing phosphorylated seryl residues in their sequence. In the present study, method for the production of CPPs from buffalo milk casein was optimised and characterised for their sequence, calcium solubilising and calcium binding activities. Response surface methodology was used to optimise the conditions for hydrolysis of buffalo casein by trypsin to obtain maximum yield of CPPs. The optimum hydrolysis conditions were as follows: hydrolysis pH 7·5, temperature 37 °C, hydrolysis time 7·0 h. Under these conditions, the experimental yield obtained was 10·04±0·24%, which is slightly lower than value predicted by the model. These CPPs were able to solubilise 1·03±0·08 mg la/mg CPPs in presence of excess phosphate and bind 0·935 mg of Ca/mg of CPPs. Eight phosphopeptides i.e. αs1-CN f (37-58) 2P; αs1-CN f (37-58) 3P; αs1-CN f (35-58) 2P; αs1-CN f (35-58) 3P; αs2-CN f (2-21) 4P; αs2-CN f (138-149) 1P; β-CN f (2-28) 4P and β-CN f (33-48) 1P were identified by LC-MS/MS which contained motif for binding of divalent minerals. The sequences of these CPPs differed from that of derived from bovine casein.

Keywords

Caseinophosphopeptidescalcium binding ability and Buffalo milk
Type
Research Article
Information
Journal of Dairy Research , Volume 81 , Issue 3 , August 2014 , pp. 364 - 371
Copyright
Copyright © Proprietors of Journal of Dairy Research 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adamson, NJ & Reynolds, EC 1995 Characterization of multiply phosphorylated peptides selectively precipitated from a pancreatic casein digest. Journal of Dairy Science 78 2653–2659Google Scholar
Adler-Nissen, J 1986 Enzymatic Hydrolysis of Food Proteins. pp. 32–35. London: Elsevier Applied Science PublishersGoogle Scholar
Allen, G 1989 Sequencing of proteins and peptides. In Laboratory Techniques in Biochemistry and Molecular Biology Volume 9, p. 73 (Eds Burdon, RH & van Knippenberg, PH), Amsterdam, Netherlands: Elsevier Science PublishersGoogle Scholar
Andrews, AT, Williams, RJH, Brownsell, VL, Isgrove, FH, Jenkins, K & Kanekanian, AD 2006 β-CN-5P and β-CN-4P components of bovine milk proteose–peptone: large scale preparation and influence on the growth of cariogenic microorganisms. Food Chemistry 96 234–241Google Scholar
Antal, J, Pal, G, Asbóth, B, Buzás, Z, Patthy, A & Gráf, L 2001 Specificity assay of serine proteinases by reverse-phase high-performance liquid chromatography analysis of competing oligopeptide substrate library. Analytical Biochemistry 288 156–167Google Scholar
Bennet, T, Desmond, A, Harrington, M, McDonagh, D, FitzGerald, RJ, Flynn, A & Cashman, KD 2000 The effect of high intakes of casein phosphopeptides on calcium absorption in the rat. British Journal of Nutrition 83 673–680Google Scholar
Berrocal, R, Chanton, S, Juillerat, MA, Pavillard, B, Scherz, JC & Jost, R 1989 Tryptic phosphopeptides from whole caseinII. Physicochemical properties related to the solubilisation of calcium. Journal of Dairy Research 56 335–341CrossRefGoogle Scholar
Corsetti, A, Massitti, O, Minervini, F, Fox, PF & Gobbetti, M 2003 Production of caseinophosphopeptides from Na-caseinates prepared from the milk of several species by a proteinase of Lactobacillus helveticus PR4. Food Biotechnology 17 183–192CrossRefGoogle Scholar
Cosenza, G, Pauciullo, A, Feligini, M, Coletta, A, Colimoro, L, Berardino, D & Ramunno, L 2009 A point mutation in the splice donor site of intron 7 in the αs2-casein encoding gene of the Mediterranean river buffalo results in an allele-specific exon skipping. Animal Genetics 40 791–794Google Scholar
Diaz, M, Dunn, CM, McClements, DJ & Decker, EA 2003 Use of caseinophosphopeptides as natural antioxidants in oil-in-water emulsions. Journal of Agricultural Food Chemistry 51 2365–2370Google Scholar
Erba, D, Ciapplellano, S & Testolin, G 2002 Effect of the ratio of casein phosphopeptides to calcium (w/w) on passive calcium transport in the distal small intestine of rats. Nutrition 18 743–746Google Scholar
Ellegard, KH, Gammelgard-Larsen, C, Sorensen, ES & Fedesov, S 1999 Process scale chromatographic isolation, characterisation and identification of tryptic bioactive casein phosphopeptides. International Dairy Journal 9 639–652Google Scholar
El - Shibiny, S, Metwally, MM, El - Dieb, SM & Assem, F 2001 Composition of glycomacropeptide produced from different casein by action of some milk clotting enzymes. Egyptian journal of immunology/Egyptian Association of Immunologists 29 217–225Google Scholar
FAO Statistics Division 2011 Buffalo Milk (whole, fresh) Production, <http://faostat.fao.org/site/569/default.aspx#ancor>>Google Scholar
Ferranti, P, Scaloni, A, Caira, S, Chianese, L, Malorni, A & Addeo, F 1998 The primary structure of water buffalo αs1- and β-Casein: identification of phosphorylation sites and characterization of a novel β-Casein variant. Journal of Protein Chemistry 17 835–844Google Scholar
Ferraretto, A, Gravaghi, C, Fiorilli, A & Tettamanti, G 2003 Casein-derived bioactive phosphopeptides: role of phosphorylation and primary structure in promoting calcium uptake by HT-29 tumor cells. FEBS Letter 551 92–98Google Scholar
Fox, KK 1970 Casein and whey protein. In Byproducts from Milk, vol. 2, pp. 341–342 (Eds Webb, BH & Whittier, EO), USA: AVI Publishing CompanyGoogle Scholar
Gerber, HW & Jost, R 1986 Casein phosphopeptides their effect on calcification of in vitro cultured embryonic rat bone. Calcified Tissue International 38 350–357Google Scholar
Hansen, M, SandstroÈm, B & LoÈnnerdal, B 1996 The effect of casein phosphopeptides on zinc and calcium absorption from high phytate infant diets assessed in rat pups and caco-2 cells. Pediatric Research 40 547–552CrossRefGoogle ScholarPubMed
Hata, I, Ueda, J & Otani, H 1999 Immunostimulatory action of a commercially available casein phosphopeptide preparation, CPP-III, in cell cultures. Milchwissenshaft 54 3–7Google Scholar
Holt, C 1996 Ability of a b-casein phosphopeptide to modulate the precipitation of calcium phosphate by forming amorphous dicalcium phosphate nanoclusters. Biochemical Journal 314 1035–1039CrossRefGoogle Scholar
Khuri, AI & Cornell, JA 1987 Response Surfaces: Designs and Analysis, pp. 12–16. New York: Marcel DekkerGoogle Scholar
Kibangou, IB, Bouhallab, S, Henry, G, Bureau, F, Allouche, S, Blais, A, Guerin, P, Arhan, P & Bougle, DL 2005 Milk Protein and Iron absorption: Contrasting effects of different caseinophosphopeptides. Pediatric Research 58 731–734CrossRefGoogle ScholarPubMed
Kitts, DD & Yuan, YV 1992 Caseinophosphopeptides and calcium bioavailability. Trends in Food Science and Technology 3 31–35CrossRefGoogle Scholar
Lin, X, Jean-Marc, S, Sarah, SC & Alain, VD 2007 Parameters that affect the identification of phosphopeptides for β-casein. Canadian Journal of Analytical Sciences and Spectroscopy 52 305–315Google Scholar
Lowry, OH, Rosebrough, NF, Farr, AL & Randall, RJ 1951 Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193 265–275Google Scholar
McDonagh, D & FitzGerald, RJ 1998 Production of Caseinophosphopeptides (CPPs) from sodium caseinate using a range of protease preparations. International Dairy Journal 8 39–45CrossRefGoogle Scholar
Miquel, E, Gomez, JA, Alegria, A, Barbera, R, Farre, R & Recio, I 2006 Identification of casein phosphopeptides after simulated gastrointestinal digestion by tandem mass spectrometry. European Food Research and Technology 222 48–53Google Scholar
Mishra, BP, Mukesh, M, Kataria, RS, Kumar, V, Pandey, D & Ahlawat, SPS 2005 Sequence analysis of beta-casein gene of Indian riverine buffalo (bubalus bubalis) http://www.uniprot.org/uniprot/Q9TSI0Google Scholar
Mukesh, M, Mishra, BP, Kataria, RS, Sobti, RC & Ahlawat, SP 2006 Sequence analysis of UTR and coding region of kappa-casein gene of Indian riverine buffalo (Bubalus bubalis). DNA Sequence 17 94–98Google Scholar
Naito, H 1990 Casein phosphopeptide: intra-intestinal formation and significance in the bioavailability of calcium. In Nutrition: Proteins and Amino Acids, pp. 207–220 (Eds Yoshida, A, Naito, H, Niiyama, Y & Suzuchi, T) Berlin: SpringerGoogle Scholar
Otani, H, Kihara, Y & Park, M 2000 The immunoenhancing property of a dietary casein phosphopeptide preparation in mice. Food and Agricultural Immunology 12 165–173Google Scholar
Perich, JW, Kelly, DP & Reynolds, EC 1992 Efficient solution-phase synthesis of phosphopeptides related to casein and stratherin. International journal of Peptide and Protein Research 40 81–88Google Scholar
Petrilli, P, Pucci, P, Pelissier, JP & Addeo, F 1987 Digestion by pancreatic juice of a beta-casomorphin-containing fragment of buffalo beta-casein. International Journal of Peptide and Protein Research 29 504–508Google Scholar
Reynolds, EC, Cain, CJ, Webber, FL, Black, CL, Riley, PF, Johnson, IH & Perich, JW 1995 Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat. Journal of Dental Research 74 1272–1279Google Scholar
Sakanaka, S, Tachibana, Y, Ishihara, N & Juneja, LR 2005 Antioxidant properties of casein calcium peptides and their effects on lipid oxidation in beef homogenates. Journal of Agricultural and Food Chemistry 53 464–468Google Scholar
Sato, R, Masanori, S, Gunshin, H, Noguchi, T & Naito, H 1991 Characterisation of phosphopeptide derived from bovine β-casein an inhibitor of intra-intestinal precipitation of calcium phosphate. Biochimica et Biophysica Acta 1077 413–441Google Scholar
Sato, R, Noguchi, T & Naito, H 1983 The necessity for the phosphate portion of casein molecules to enhance Ca absorption from the small intestine. Agricultural Biological Chemistry 47 2415–2417Google Scholar
Schlimme, E & Meisel, H 1993 Bioactive peptides: structural, physiological and analytical aspects. Newsletter of the IDF 139 57–89Google Scholar
Schmidt, DG, Both, P, Visser, S, Slangen, KJ & van Rooijen, PJ 1987 Studies on the precipitation of calcium phosphate. II Experiments in the pH range 7·3 to 5·6 at 25 and 50 °C in the presence of additives. Netherlands Milk and Dairy Journal 41 121–136Google Scholar
Su, R, Qi, W, He, Z, Yuan, S & Zhang, Y 2007 Pancreatic hydrolysis of bovine casein: identification and release kinetics of phosphopeptides. Food Chemistry 104 276–286Google Scholar
Sukla, S, Bhattacharya, TK, Venkatachalapathy, RT, Kumar, P & Sharma, A 2006 Cloning and characterization of sigma2-casein gene of Riverine buffalo. DNA Sequence 17 458–464Google Scholar
Sukla, S, Bhattacharya, TK, Venkatachalapathy, RT, Kumar, P & Sharma, A 2007 Molecular cloning and characterization of buffalo alpha(s1)-casein gene. DNA Sequence 18 334–340Google Scholar
Tsuchita, H, Goto, T, Shimizu, T, Yonehara, Y & Kuwata, T 1996 Dietary casein phosphopeptides prevent bone loss in aged ovariectomised rats. Journal of Nutrition 126 86–93Google Scholar
Wei, Q, Zhimin, H & Deqing, S 2003 Product distribution of casein tryptic hydrolysis based on HPLEC analysis and molecular mechanism. Chemical Engineering Science 58 767–775Google Scholar
Xu, C, Lu, Y, Ma, J, Mohammadi, M & Neubert, TA 2005 Identification of Phosphopeptides by MALDI Q-TOF MS in positive and negative ion modes after methyl esterification. Molecular & Cellular Proteomics 4 809–818Google Scholar
Yoshikawa, M, Sasaki, R & Chiba, H 1981 Effects of chemical Phosphorylation of Bovine Casein components on the properties related to Casein Micelle formation. Agricultural and Biological Chemistry 45 909–914Google Scholar
Zhu, YS & FitzGerald, RJ 2010 Direct nano HPLC–ESI–QTOF MS/MS analysis of tryptic caseinophosphopeptides. Food Chemistry 123 753–759CrossRefGoogle Scholar