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Isolation of a calcium-binding peptide from bovine serum protein hydrolysates

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Abstract

A calcium-binding peptide was isolated from the hydrolysates of bovine serum protein (BSP). BSP was hydrolyzed using 3 different types of proteases, Alcalase, Flavourzyme, and Protamex, and the degree of hydrolysis was determined and monitored using trinitrobenzenesulfonic acid and SDS-PAGE. The hydrolysates of BSP using Alcalase were selected and ultra-filtered below 3 kDa. The membrane-filtered solution was then fractionated using ion exchange chromatography and normal phase HPLC to isolate a calcium-binding peptide. The calcium-binding capacity was determined by the orthophenanthroline method. The sequence of the purified calcium-binding peptide was analyzed using LC/electron spray ionization (LC/ESI)-tandem mass spectroscopy and identified to be Asp-Asn-Leu-Pro-Asn-Pro-Glu-Asp-Arg-Lys-Asn-Tyr-Glu, which has a molecular weight of 1,603 Da.

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References

  1. Jin YG, Fu WW, Ma MH. Preparation and structure characterization of soluble bone collagen peptide chelating calcium. Afr. J. Biotechnol. 10: 10204–10211 (2011)

    CAS  Google Scholar 

  2. Bass J, Chan G. Calcium nutrition and metabolism during infancy. Nutrition 22: 1057–1066 (2006)

    Article  CAS  Google Scholar 

  3. Lee SH, Song KB. Isolation of a calcium-binding peptide from enzymatic hydrolysates of porcine blood plasma protein. J. Korean Soc. Appl. Biol. Chem. 52: 290–294 (2009)

    Article  CAS  Google Scholar 

  4. Silva SV, Malcata FX. Caseins as source of bioactive peptides. Int. Dairy J. 15: 1–15 (2005)

    Article  CAS  Google Scholar 

  5. Megias C, Pedroche J, Yust MM, Giron-Calle J, Alaiz M, Millan F, Vioque J. Affinity purification of copper-chelating peptides from sunflower protein hydrolysates. J. Agr. Food Chem. 55: 6509–6514 (2007)

    Article  CAS  Google Scholar 

  6. Tessier B, Harscoat-Schiavo C, Marc I. Selective separation of peptides contained in a rapeseed (Brassica campestris L.) protein hydrolysate using UF/NF membranes. J. Agr. Food Chem. 54: 3578–3584 (2006)

    Article  CAS  Google Scholar 

  7. Rhee MS, Kim BC. Effect of low voltage electrical stimulation and temperature conditioning on postmortem changes in glycolysis and calpains activities of Korean native cattle (hanwoo). Meat Sci. 58: 231–237 (2001)

    Article  CAS  Google Scholar 

  8. Bizzotto CS, Capobiango M, Silvestre MPC. Evaluation of functional properties of a blood protein. Pakistan J. Nutr. 4: 11–16 (2005)

    Article  Google Scholar 

  9. Duarte RT, Simoes MCC, Sgarbieri VC. Bovine blood components: Fractionation, composition, and nutritive value. J. Agr. Food Chem. 47: 231–236 (1999)

    Article  CAS  Google Scholar 

  10. Hyun CK, Shin HK. Production of angiotensin I convertion enzyme inhibitory peptides from bovine blood plasma proteins. Korean J. Biotechnol. Bioeng. 14: 600–605 (1999)

    Google Scholar 

  11. Lee SH, Song KB. Isolation of an angiotensin converting enzyme inhibitory peptide from irradiated bovine blood plasma protein hydrolysates. J. Food Sci. 68: 2469–2472 (2003)

    Article  CAS  Google Scholar 

  12. Elkund A. On the determination of available lysine in casein and rapeseed protein concentration using 2,4,6-trinitronbenzensulphonic acid (TNBS) as a reagent of free ɛ-amino group of lysine. Anal. Chem. 70: 434–439 (1976)

    Google Scholar 

  13. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970)

    Article  CAS  Google Scholar 

  14. Gitelman HJ. An improved automated procedure for the determination of calcium in biological specimens. Anal. Biochem. 18: 521–531 (1967)

    Article  CAS  Google Scholar 

  15. Korhonen H, Pihlanto A. Bioactive peptides: Production and functionality. Int. Dairy J. 16: 945–960 (2006)

    Article  CAS  Google Scholar 

  16. Korhonen H, Pihlanto A. Bioactive peptides: Novel applications for milk proteins. Appl. Biotech. Food Sci. Policy 1: 133–144 (2003)

    CAS  Google Scholar 

  17. Jung WK, Lee BJ, Kim SK. Fish-bone peptide increases calcium solubility and bio availability in ovariectomised rats. Brit. J. Nutr. 95: 124–128 (2006)

    Article  CAS  Google Scholar 

  18. Vegarud GE, Langsrud T, Svenning C. Mineral-binding milk proteins and peptides; occurrence, biochemical, and technological characteristics. Brit. J. Nutr. 84: 91–98 (2000)

    Article  Google Scholar 

  19. Sun H, Li H, Sadler PJ. Transferrin as a metal ion mediator. Chem. Rev. 99: 2817–2842 (1999)

    Article  CAS  Google Scholar 

  20. Korhonen H, Pihlanto A. Bioactive peptides and proteins. Vol. 47, pp. 175–276. In: Advances in Food and Nutrition Research. Taylor SL (ed). Academic Press, San Diego, CA, USA (2003)

    Google Scholar 

  21. Kim SB, Lim JW. Calcium-binding peptides derived from tryptic hydrolysates of cheese whey protein. Asian-Aust. J. Anim. Sci. 17: 1459–1464 (2004)

    CAS  Google Scholar 

  22. Cariolou MA, Morse DE. Purification and characterization of calcium-binding conchiolin shell peptides from the mollusk, Haliotes rufescens, as a function of development. J. Comp. Physiol. B. 157: 717–729 (1988)

    Article  CAS  Google Scholar 

  23. Nemirovskiy OV, Gross ML. Intrinsic Ca2+ affinities of peptides: Application of the kinetic method to analogs of calcium-binding site ø of rabbit skeletal troponin C. Am. Soc. Mass Spectrom. 11: 770–779 (2000)

    Article  CAS  Google Scholar 

  24. Tuan RS, Scott WA, Cohn ZA. Purification and characterization of calcium-binding protein from chick chorioallantoic membrane. J. Biol. Chem. 253: 1011–1016 (1978)

    CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Food Science and Technology, Chungnam National University, Daejeon, 305-764, Korea

    Dong-Won Choi, Ji-Hye Lee, Ho-Hyun Chun & Kyung Bin Song

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  1. Dong-Won Choi
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  2. Ji-Hye Lee
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  3. Ho-Hyun Chun
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  4. Kyung Bin Song
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Correspondence to Kyung Bin Song.

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Choi, DW., Lee, JH., Chun, HH. et al. Isolation of a calcium-binding peptide from bovine serum protein hydrolysates. Food Sci Biotechnol 21, 1663–1667 (2012). https://doi.org/10.1007/s10068-012-0221-z

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  • DOI: https://doi.org/10.1007/s10068-012-0221-z

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