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

Published online by Cambridge University Press:  16 January 2015

Shun-Li Huang ,
Li-Na Zhao ,
Xixi Cai ,
Shao-Yun Wang ,
Yi-Fan Huang ,
Jing Hong  and
Ping-Fan Rao
Shun-Li Huang
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
Li-Na Zhao
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
Xixi Cai
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
Shao-Yun Wang*
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
Yi-Fan Huang
Affiliation:
College of Food Science, Fujian Agriculture and Forestry University, No.15 Shangxiadian Road, Fujian, Fuzhou 350002, China
Jing Hong
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
Ping-Fan Rao
Affiliation:
College of Bioscience and Biotechnology, Fuzhou University, No.2 Xueyuan Road, Fujian, Fuzhou 350002, China
*
*For correspondence; e-mail: shywang@fzu.edu.cn
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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.

Keywords

Whey protein hydrolysiscalcium-binding peptideseparation
Type
Research Article
Information
Journal of Dairy Research , Volume 82 , Issue 1 , February 2015 , pp. 29 - 35
Copyright
Copyright © Proprietors of Journal of Dairy Research 2015 

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References

Armas, A, Sonois, V, Mothes, E, Zazarguil, H & Faller, P 2006 Zinc binds to the neuroprotective peptide humanin. Journal of Inorganic Biochemistry 100 1672–1678Google Scholar
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 83 673–680Google Scholar
Bronner, F & Pansu, D 1998 Nutrition aspects of calcium absorption. Journal of Nutrient 129 9–12CrossRefGoogle Scholar
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 50 871–877CrossRefGoogle ScholarPubMed
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 5 689–697Google Scholar
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 3030–3038CrossRefGoogle Scholar
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 232 967–981Google Scholar
Feng, M, van der Does, L & Bantjes, A 1995 Preparation of apolactoferrin with a very low iron saturation. Journal of Dairy Science 78 2352–2357Google Scholar
Gitelman, HJ 1967 An improved automated procedure for the determination of calcium in biological specimens. Analytical Biochemistry 18 521–531Google Scholar
Guénguen, L & Pointillart, A 2000 The bioavailability of dietary calcium. Journal of the American College of Nutrition 19 119–136Google Scholar
Houser, RP, Fitzsimons, MP & Barton, JK 1999 Metal-dependent intramolecular chiral induction: the Zn2+ Complex of an ethidium–peptide conjugate. Inorganic Chemistry 38 1368–1370CrossRefGoogle ScholarPubMed
Huang, GR, Ren, ZR & Jiang, JX 2011 Separation of iron-binding peptides from shrimp processing by-products hydrolysates. Food and Bioprocess Technology 4 1527–1532Google Scholar
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 80–85Google Scholar
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 91 333–340Google Scholar
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 1514–1525CrossRefGoogle ScholarPubMed
Kim, SB & Lim, JW 2004 Calcium-binding peptides derived from tryptic hydrolysates of cheese whey protein. Asian Australasian Journal of Animal Sciences 17 1459–1464Google Scholar
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 17 712–718Google Scholar
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 52 290–294Google Scholar
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 57–63Google Scholar
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 11 770–779CrossRefGoogle ScholarPubMed
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 54 365–381CrossRefGoogle ScholarPubMed
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 413–415Google Scholar
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) 28 533–541CrossRefGoogle Scholar
Vegarud, GE, Langsrud, T & Svenning, C 2000 Mineral-binding milk proteins and peptides; occurrence, biochemical and technological characteristics. British Journal of Nutrition 84 S91–S98Google Scholar
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 2731–2740Google Scholar
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 103 657–665Google Scholar
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 48 435–441Google Scholar
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 4230–4423Google Scholar