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Purification and characterisation of a glutamic acid-containing peptide with calcium-binding capacity from whey protein hydrolysate

  • Shun-Li Huang (a1), Li-Na Zhao (a1), Xixi Cai (a1), Shao-Yun Wang (a1), Yi-Fan Huang (a2), Jing Hong (a1) and Ping-Fan Rao (a1)...

Abstract

The bioavailability of dietary ionised calcium is affected by intestinal basic environment. Calcium-binding peptides can form complexes with calcium to improve its absorption and bioavailability. The aim of this study was focused on isolation and characterisation of a calcium-binding peptide from whey protein hydrolysates. Whey protein was hydrolysed using Flavourzyme and Protamex with substrate to enzyme ratio of 25 : 1 (w/w) at 49 °C for 7 h. The calcium-binding peptide was isolated by DEAE anion-exchange chromatography, Sephadex G-25 gel filtration and reversed phase high-performance liquid chromatography (RP-HPLC). A purified peptide of molecular mass 204 Da with strong calcium binding ability was identified on chromatography/electrospray ionisation (LC/ESI) tandem mass spectrum to be Glu-Gly (EG) after analysis and alignment in database. The calcium binding capacity of EG reached 67·81 μg/mg, and the amount increased by 95% compared with whey protein hydrolysate complex. The UV and infrared spectrometer analysis demonstrated that the principal sites of calcium-binding corresponded to the carboxyl groups and carbonyl groups of glutamic acid. In addition, the amino group and peptide amino are also the related groups in the interaction between EG and calcium ion. Meanwhile, the sequestered calcium percentage experiment has proved that EG-Ca is significantly more stable than CaCl2 in human gastrointestinal tract in vitro. The findings suggest that the purified dipeptide has the potential to be used as ion-binding ingredient in dietary supplements.

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Corresponding author

*For correspondence; e-mail: shywang@fzu.edu.cn

References

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Armas, A, Sonois, V, Mothes, E, Zazarguil, H & Faller, P 2006 Zinc binds to the neuroprotective peptide humanin. Journal of Inorganic Biochemistry 100 16721678
Bennett, T, Desmond, A, Harrington, M, McDonagh, D, FitzGerald, R, Flynn, A & Cashman, KD 2000 The effect of high intakes of casein and casein phosphopeptide on calcium absorption in the rat. British Journal of Nutrition 83 673680
Bronner, F & Pansu, D 1998 Nutrition aspects of calcium absorption. Journal of Nutrient 129 912
Chaud, MV, Izumi, C, Nahaal, Z, Shuhama, T, Lourdes Pires Bianchi, M & Freitas, O 2002 Iron derivatives from casein hydrolysates as a potential source in the treatment of iron deficiency. Journal of Agricultural and Food Chemistry 50 871877
Chen, D, Liu, ZY, Huang, WQ, Zhao, YH, Dong, SY & Zeng, MY 2013 Purification and characterisation of a zinc-binding peptide from oyster protein hydrolysate. Journal of Functional Foods 5 689697
Cilla, A, Lagarda, MJ, Alegría, A, Ancos, BD, Cano, MP, Sánchez-Moren, C, Plaza, L & Barberá, R 2011 Effect of processing and food matrix on calcium and phosphorous bioavailability from milk-based fruit beverages in Caco-2 cells. Food Research International 44 30303038
Fabian, HC, Schultz, D, Naumann, D, Landt, O, Hahn, U & Saenger, W 1993 Secondary structure and temperature-induced unfolding and refolding of ribonuclease-T1 in aqueous solution-a fourier transform infrared spectroscopy study. Journal of Molecular Biology 232 967981
Feng, M, van der Does, L & Bantjes, A 1995 Preparation of apolactoferrin with a very low iron saturation. Journal of Dairy Science 78 23522357
Gitelman, HJ 1967 An improved automated procedure for the determination of calcium in biological specimens. Analytical Biochemistry 18 521531
Guénguen, L & Pointillart, A 2000 The bioavailability of dietary calcium. Journal of the American College of Nutrition 19 119136
Houser, RP, Fitzsimons, MP & Barton, JK 1999 Metal-dependent intramolecular chiral induction: the Zn2+ Complex of an ethidium–peptide conjugate. Inorganic Chemistry 38 13681370
Huang, GR, Ren, ZR & Jiang, JX 2011 Separation of iron-binding peptides from shrimp processing by-products hydrolysates. Food and Bioprocess Technology 4 15271532
Jeyarajah, S & Allen, JC 1994 Calcium binding and salt-induced structural changes of native and preheated β-lactoglobulin. Journal of Agricultural and Food Chemistry 42 8085
Jung, WK, Park, PJ, Byun, HG, Moon, SH & Kim, SK 2005 Preparation of hoki (Johnius belengerii) bone oligophosphopeptide with a high affinity to calcium by carnivorous crude proteinase. Food Chemistry 91 333340
Kállay, C, Várnagy, K, Micera, G, Sanna, D & Sóvág, I 2005 Copper(II) complexes of oligopeptides containing aspartyl and glutamyl residues. Potentiometric and spectroscopic studies. Journal of Inorganic Biochemistry 99 15141525
Kim, SB & Lim, JW 2004 Calcium-binding peptides derived from tryptic hydrolysates of cheese whey protein. Asian Australasian Journal of Animal Sciences 17 14591464
Kim, SB, Shin, HS & Lim, JW 2004 Separation of calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. Asian-Aust. Journal of Animal Science 17 712718
Lee, SH & Song, KB 2009 Isolation of a calcium-binding peptide from enzymatic hydrolysates of porcine blood plasma protein. Journal of Korean Society for Applied Biological Chemistry 52 290294
Narin, C, Benjamas, C, Nualpun, S & Wirote, Y 2013 Calcium-binding peptides derived from tilapia (Oreochromis niloticus) protein hydrolysate Eur. European Food Research and Technology 236 5763
Nemirovskiy, OV & Gross, ML 2000 Intrinsic Ca2+ affinities of peptides: application of the kinetic method to analogs of calcium-binding site III of rabbit skeletal troponin C. Journal of the American Society for Mass Spectrometry 11 770779
Osborne, CG, McTyre, RB, Dudek, J, Roche, KE, Scheuplein, R, Silverstein, B, Weinberg, MS & Salkeld, AA 1996 Evidence for the relationship of calcium to blood pressure. Nutrition Reviews 54 365381
Sato, R, Shindo, M, Gunshin, H, Noguchi, T & Naito, H 1991 Characterization of phosphopeptide derived from bovine β-casein: an inhibitor to intra-intestinal precipitation of calcium phosphate. Biochimica et Biophysica Acta 1077 413415
Tamura, M, Oku, T & Hosoya, N 1982 Calcium-binding proteins in bovine milk: calc- ium-binding properties and amino acid composition. Journal Nutritional Science and Vitaminology (Tokyo) 28 533541
Vegarud, GE, Langsrud, T & Svenning, C 2000 Mineral-binding milk proteins and peptides; occurrence, biochemical and technological characteristics. British Journal of Nutrition 84 S91S98
Wang, X, Zhou, J, Tong, PS & Mao, XY 2011 Zinc-binding capacity of yak casein hydrolysate and the zinc-releasing characteristics of casein hydrolysate–zinc complexes. Journal of Dairy Science 94 27312740
Wang, XL, Li, K, Yang, XD, Wang, LL & Shen, RF 2009 Complexation of Al(III) with reduced glutathione in acidic aqueous solutions. Journal of Inorganic Biochemistry 103 657665
Wu, HH, Liu, ZY, Zhao, YH & Zeng, MY 2012 Enzymatic preparation and characterization of iron-chelating peptides from anchovy (Engraulis japonicus) muscle protein. Food Research International 48 435441
Zhou, J, Wang, X, Ai, T, Cheng, X, Guo, HY, Teng, GX & Mao, XY 2012 Preparation and characterization of β-lactoglobulin hydrolysate–iron complexes. Journal of Dairy Science 95 42304423

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