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Isolation of a Calcium-binding Peptide from Chlorella Protein Hydrolysates

  • Jeon, So-Jeong (Department of Food Science and Technology, Chungnam National University) ;
  • Lee, Ji-Hye (Department of Food Science and Technology, Chungnam National University) ;
  • Song, Kyung-Bin (Department of Food Science and Technology, Chungnam National University)
  • Received : 2010.10.08
  • Accepted : 2010.11.26
  • Published : 2010.12.31

Abstract

To isolate a calcium-binding peptide from chlorella protein hydrolysates, chlorella protein was extracted and hydrolyzed using Flavourzyme, a commercial protease. The degree of hydrolysis and calcium-binding capacity were determined using trinitrobenzenesulfonic acid and orthophenanthroline methods, respectively. The enzymatic hydrolysis of chlorella protein for 6 hr was sufficient for the preparation of chlorella protein hydrolysates. The hydrolysates of chlorella protein were then ultra-filtered under 5 kDa as molecular weight. The membrane-filtered solution was fractionated using ion exchange, reverse phase, normal phase chromatography, and fast protein liquid chromatography to identify a calcium-binding peptide. The purified calcium-binding peptide had a calcium binding activity of 0.166 mM and was determined to be 700.48 Da as molecular weight, and partially identified as a peptide containing Asn-Ser-Gly-Cys based on liquid chromatography/electrospray ionization tandem mass spectrum.

Keywords

References

  1. Grigorova S. 2005. Dry biomass of fresh water algae ofchlorella genus in the combined forages for laying hens.J Cent Eur Agric 6: 625-630.
  2. Huang Z, Li L, Huang G, Yan Q, Shi B, Xu X. 2009.Growth-inhibitory and metal-binding proteins in Chlorellavulgaris exposed to cadmium or zinc. Aquat Toxicol 91:54-61. https://doi.org/10.1016/j.aquatox.2008.10.003
  3. Hasegawa T, Kimura Y, Hiromatsu K, Kobayashi N,Yamada A, Makino M, Okuda M, Sano T, Nomoto K,Yoshikai Y. 1997. Effect of hot water extract of Chlorellavulgaris on cytokine expression patterns in mice with murineacquired immunodeficiency syndrome after infectionwith Listeria monocytogenes. Immunopharm 35: 273-282. https://doi.org/10.1016/S0162-3109(96)00150-6
  4. Hasegawa T, Ito K, Kumamoto S, Ando Y, Yamada A,Nomoto K, Yasunobu Y. 1999. Oral administration of hotwater extracts of Chlorella vulgaris reduces IgE productionagainst milk casein in mice. Int J Immunopharmacol21: 311-323. https://doi.org/10.1016/S0192-0561(99)00013-2
  5. Janczyk P, Franke H, Souffrant WB. 2007. Nutritional valueof Chlorella vulgaris: Effects of ultrasonication andelectroporation on digestibility in rats. Anim Feed Sci Tech132: 163-169. https://doi.org/10.1016/j.anifeedsci.2006.03.007
  6. Rollemberg MC, Simoes Goncalves MLS, Correia dosSantos MM, Botelho MJ. 1999. Thermodynamics of uptakeof cadmium by Chlorella marina. BioelectrochemBioenerg 48: 61-68. https://doi.org/10.1016/S0302-4598(98)00220-7
  7. Suetsuna K, Chen JR. 2001. Identification of antihypertensivepeptides from peptic digest of two microalgae,Chlorella vulgaris and Spirulina platensis. Mar Biotechnol3: 305-309. https://doi.org/10.1007/s10126-001-0012-7
  8. Kang MS, Chae HJ. 2003. Biological efficacy assay ofChlorella hydrolysates. J Korean Acad Ind Coop Soc 4:366-371.
  9. Sheih IC, Fang TJ, Wu TK, Lin PH. 2010. Anticancerand antioxidant activities of the peptide fraction from algaeprotein waste. J Agric Food Chem 58: 1202-1207. https://doi.org/10.1021/jf903089m
  10. Cha JY, Kim JW, Park BK, Jin HJ, Kim SY, Cho YS.2008. Isolation and identification of Chlorella sp. CMS-1and the chemical composition of its hot water extract. JLife Sci 18: 1723-1727. https://doi.org/10.5352/JLS.2008.18.12.1723
  11. Iwamoto H. 2003. Industrial production of microalgal cell -mass and secondary products- major industrial species:Chlorella. In Handbook of Microalgal Culture: Biotechnologyand Applied Phycology. Richmond A, ed.Blackwell Publishing, Oxford, England. p 255-263.
  12. Morris HJ, Almarales A, Carrillo O, Bermudez RC. 2008.Utilization of Chlorella vulgaris cell biomass for the productionof enzymatic protein hydrolysates. BioresourceTechnol 99: 7723-7729. https://doi.org/10.1016/j.biortech.2008.01.080
  13. Sander K, Murthy GS. 2009. Enzymatic degradation ofmicroalgal cell walls. An ASABE Meeting Presentation.Paper No. 1035636.
  14. Morris HJ, Carrillo O, Almarales A, Bermudez RC,Lebeque Y, Fontaine F, Llaurado G, Beltran Y. 2007.Immunostimulant activity of an enzymatic protein hydrolysatefrom green microalgae Chlorella vulgaris on undernourishedmice. Enzyme Microb Tech 40: 456-460. https://doi.org/10.1016/j.enzmictec.2006.07.021
  15. Rui XU. 2009. Calcium binding of peptides derived fromenzymatic hydrolysates of whey protein concentrate. IntJ Dairy Technol 62: 170-173. https://doi.org/10.1111/j.1471-0307.2009.00477.x
  16. Hutyra T, Iwanczak B. 2009. Determination of milk anddairy products consumption and their connection with lactosemalabsorption or lactose intolerance in selected disordersof the alimentary tract in children. Pol Merkur Lekarski 26: 110-116.
  17. Bass J, Chan G. 2006. Calcium nutrition and metabolismduring infancy. Nutrition 22: 1057-1066. https://doi.org/10.1016/j.nut.2006.05.014
  18. Lee SH, Song KB. 2009. Isolation of a calcium-bindingpeptide from enzymatic hydrolysates of porcine bloodplasma protein. J Korean Soc Appl Biol Chem 52: 290-294. https://doi.org/10.3839/jksabc.2009.051
  19. Silva SV, Malcata FX. 2005. Casein as source of bioactivepeptides. Int Dairy J 15: 1-15. https://doi.org/10.1016/j.idairyj.2004.04.009
  20. Lee SH, Song KB. 2009. Purification of an iron-bindingnona-peptide from hydrolysates of porcine blood plasmaprotein. Process Biochem 44: 378-381. https://doi.org/10.1016/j.procbio.2008.12.001
  21. Eklund A. 1976. On the determination of available lysinein casein and rapeseed protein concentration using 2,4,6-trinitrobenzene-sulphonic acid (TNBS) as a reagent of free${\alpha}-amino$ group of lysine. Anal Chem 70: 434-439.
  22. Han XQ, Parkin KL, Lincourt RH, Gao S. 2002. Isolatedantioxidant peptides. US Patent 006465432B1.
  23. Gitelman HJ. 1967. An improved automated procedure forthe determination of calcium in biological specimens. AnalBiochem 18: 521-531. https://doi.org/10.1016/0003-2697(67)90110-8
  24. SAS Institute. 2001. SAS system for windows. Version 8.2,SAS Institute, Inc., Cary, NC, USA.
  25. Korhonen H, Pihlanto A. 2006. Bioactive peptides:Production and functionality. Int Dairy J 16: 945-960. https://doi.org/10.1016/j.idairyj.2005.10.012
  26. Bourseau P, Vandanjon L, Jaouen P, Chaplain-DerouiniotM, Masse A, Guerard F, Chabeaud A, Fouchereau-PeronM, Le Gal Y, Ravallec-Ple R, Berge JP, Picot L, PiotJM, Batista I, Thorkelsson G, Delannoy C, Jakobsen G,Johansson I. 2009. Fractionation of fish protein hydrolysatesby ultrafiltration and nanofiltration: impact on peptidicpopulations. Desalination 244: 303-320. https://doi.org/10.1016/j.desal.2008.05.026
  27. Hara H, Funabiki R, Iwata M, Yamazaki KI. 1984. Portalabsorption of small peptides in rats under unrestrainedconditions. J Nutr 114: 1122-1129.
  28. Shimizu M. 2004. Food-derived peptides and intestinalfunctions. BioFactors 21: 43-47. https://doi.org/10.1002/biof.552210109
  29. Hartmann R, Meisel H. 2007. Food-derived peptides withbiological activity: from research to food applications.Curr Opin Biotech 18: 163-169. https://doi.org/10.1016/j.copbio.2007.01.013
  30. Miquel E, Alegria A, Barbera R, Farre R. 2005. Speciationanalysis of calcium, iron, and zinc in casein phosphopeptidefractions from toddler milk-based formula by anionexchange and reversed-phased high-performance liquidchromatography-mass spectrometry/frame atomic-absorptionspectroscopy. Anal Bioanal Chem 381: 1082-1088. https://doi.org/10.1007/s00216-004-3002-6
  31. Jung WK, Karawita R, Heo SJ, Lee BJ, Kim SK, JeonYJ. 2006. Recovery of a novel Ca-binding peptide fromAlaska Pollack (Theragra chalcogramma) backbone bypepsinolytic hydrolysis. Process Biochem 41: 2097-2100. https://doi.org/10.1016/j.procbio.2006.05.008
  32. Maliarik MJ, Roberts DD, Goldstein IJ. 1991. Antigenicand calcium binding properties of a peptide containing theessential cysteine in lima bean lectin. Plant Physiol 95:286-290. https://doi.org/10.1104/pp.95.1.286

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