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Sequence Determination of an Antioxidant Peptide Obtained by Enzymatic Hydrolysis of Oyster Crassostrea madrasensis (Preston)

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Abstract

Soft tissue from cultured farm fresh oysters (Crassostrea madrasensis) was subjected to two standard enzymatic peptide extraction procedures using pepsin and papain. The crude extracts obtained were partially purified by column chromatography and were freeze-dried. The hydrolysates obtained were compared with respect to their degree of hydrolysis (DH), antioxidant potential (AP) and total phenolic content (TPC). The hydrolysate showing better antioxidant property was further subjected to purification by high performance liquid chromatography and characterized by LC-MS/MS. Papain-digested oyster protein (OPHpap) hydrolysate showed higher DH, AP and TPC. OPHpap was further subjected to ultrafiltration and fractionated into 3 sizes namely, above 10, 3–10 and 1–3 kDa according to the molecular size. Antioxidant capacity of <3 kDa fraction OPHpap-3 evaluated by DPPH free radical scavenging assay, metal chelating activity, linoleic acid autoxidation assay showed maximum effectiveness. Of the seven fractions collected by purification of OPH-pap-3 on semi-preparative RP-HPLC, fraction 7 that showed the highest antioxidant activity was further characterized by LC-ESI-MS/MS and its sequence determined. An antioxidant peptide molecule with thirteen amino acids was identified in oyster protein hydrolysate obtained by papain digestion that may find application as a nutraceutical or may be utilized in food industry for prevention of rancidity in foods.

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Notes

  1. http://peptide2.com/N_peptide_hydrophobicity_hydrophilicity.php.

References

  • Anderson RS, Beaven AE (2001) Antibacterial activities of oyster (Crassostrea virginica) and mussel (Mytilus edulis and Geukensia demissa) plasma. Aquat Living Resour 14:343–349

    Article  Google Scholar 

  • Asha KK, Anandan R, Mathew S, Lakshmanan PT (2014) Biochemical profile of oyster Crassostrea madrasensis and its nutritional attributes. Egypt J Aquat Res 40:35–41

    Article  Google Scholar 

  • Bougatef A, Hajji M, Balti R, Lassoued I, Triki-Ellouz Y, Nasri M (2009) Antioxidant and free radical-scavenging activities of smooth hound (Mustelus mustelus) muscle protein hydrolysates obtained by gastrointestinal proteases. Food Chem 114:1198–1205

    Article  CAS  Google Scholar 

  • Bougatef A, Nedjar-Arroume N, Manni L, Ravallec R, Barkia A, Guillochon D, Nasri M (2010) Purification and identification of novel antioxidant peptides from enzymatic hydrolysates of sardinelle (Sardinella aurita) by-products proteins. Food Chem 118:559–565

    Article  CAS  Google Scholar 

  • Chay Pak Ting BP, Mine Y, Juneja LR, Okubo T, Gauthier SF, Pouliot Y (2001) Comparative composition and antioxidant activity of peptide fractions obtained by ultrafiltration of egg yolk protein enzymatic hydrolysates. Membranes 1:149–161

    Article  Google Scholar 

  • Cheng Z, Ren J, Li Y, Chang W, Chen Z (2003) Establishment of a quantitative structure-activity relationship model for evaluating and predicting the protective potentials of phenolic antioxidants on lipid peroxidation. J Pharm Sci 92:475–484

    Article  CAS  PubMed  Google Scholar 

  • Cinq-Mars CD, Hu C, Kitts DD, Li-Chan EC (2008) Investigations into inhibitor type and mode, simulated gastrointestinal digestion, and cell transport of the angiotensin I-converting enzyme-inhibitory peptides in Pacific hake (Merluccius productus) fillet hydrolysate. J Agric Food Chem 56:410–419

    Article  CAS  PubMed  Google Scholar 

  • Dinis TCP, Madeira VMC, Almeida LM (1994) Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch of Biochem Biophys 315:161–169

    Article  CAS  Google Scholar 

  • Dong SY, Zeng MY, Wang DF, Liu ZY, Zhao YH, Yang HC (2008) Antioxidant and biochemical properties of protein hydrolysates prepared from Silver carp (Hypophthalmichthys molitrix). Food Chem 107:1485–1493

    Article  CAS  Google Scholar 

  • Fruton JS, Fujii S, Knappenberger MH (1961) The mechanism of pepsin action. Proc Natl Acad Sci USA 15(47):759–761

    Article  Google Scholar 

  • Halliwell B, Grootveld M, Gutteridge JM (1988) Methods for the measurement of hydroxyl radicals in biomedical systems: deoxyribose degradation and aromatic hydroxylation. Methods Biochem Anal 33:59–90

    Article  CAS  PubMed  Google Scholar 

  • Hsu KC (2010) Purification of antioxidative peptides prepared from enzymatic hydrolysates of tuna dark muscle by-product. Food Chem 122:42–48

    Article  CAS  Google Scholar 

  • Ichikawa K, Okabayashi T, Shima Y, Iiyama T, Takezaki Y, Munekage M, Namikawa T, Sugimoto T, Kobayashi M, Mimura T, Hanazaki K (2012) Branched-chain amino acid-enriched nutrients stimulate antioxidant DNA repair in a rat model of liver injury induced by carbon tetrachloride. Mol Biol Rep 39:10803–10810

    Article  CAS  PubMed  Google Scholar 

  • Iris F, Benzie F, Strain JJ (1999) Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. In: Packer L (ed) Methods in enzymology. Academic Press, New York, pp 15–27

    Google Scholar 

  • Ishida Y, Fujita T, Asai K (1981) New detection and separation method for amino acids by high-performance liquid chromatography. J Chromatogr 204:143–148

    Article  CAS  PubMed  Google Scholar 

  • Je JY, Park PJ, Kim SK (2005) Antioxidant activity of a peptide isolated from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate. Food Res Int 38:45–50

    Article  CAS  Google Scholar 

  • Kim SY, Je JY, Kim SK (2007) Purification and characterization of antioxidant peptide from hoki (Johnius belengerii) frame protein by gastrointestinal digestion. J Nutri Biochem 18:31–38

    Article  CAS  Google Scholar 

  • Klompong V, Benjakul S, Kantachote D, Hayes KD, Shahidi F (2008) Comparative study on antioxidative activity of yellow stripe trevally protein hydrolysate produced from Alcalase and Flavourzyme. Int J Food Sci Technol 43:1019–1026

    Article  CAS  Google Scholar 

  • Klompong V, Benjakul S, Yachai M, Visessanguan W, Shahidi F, Hayes K (2009) Amino acid composition and antioxidative peptides from protein hydrolysates of yellow stripe trevally (Selaroides leptolepis). J Food Sci 74:C126–C133

    Article  CAS  PubMed  Google Scholar 

  • Kumar NSS, Nazeer RA, Jaiganesh R (2011) Purification and biochemical characterization of antioxidant peptide from horse mackerel (Magalaspis cordyla) viscera protein. Peptides 32:1496–1501

    Article  Google Scholar 

  • Levine RL, Bertlet BS, Moskovitz J, Mosoni L (1999) Methionine residues may protect proteins from critical oxidative damage. Mech Ageing Dev 107:323–332

    Article  CAS  PubMed  Google Scholar 

  • Li Y-W, Li B (2013) Characterization of structure-antioxidant activity relationship of peptides in free radical systems using QSAR models: key sequence positions and their amino acid properties. J Theor Biol 318:29–43

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, Dong S, Xu J, Zeng M, Song H, Zhao Yuanhui (2008) Production of cysteine-rich antimicrobial peptide by digestion of oyster (Crassostrea gigas) with alcalase and bromelin. Food Control 19:231–235

    Article  Google Scholar 

  • Maralani MN, Movahedian A, Javanmard SH (2012) Antioxidant and cytoprotective effects of l-serine on human endothelial cells. Res Pharm Sci 7:209–215

    PubMed  PubMed Central  Google Scholar 

  • Marques C, Licks F, Zattoni I, Borges B, de Souza LER, Marroni CA, Marroni NP (2013) Antioxidant properties of glutamine and its role in VEGF-Akt pathways in portal hypertension gastropathy. World J Gastroenterol 19:4464–4474

    Article  PubMed  PubMed Central  Google Scholar 

  • Mendis E, Rajapakse N, Kim SK (2004) Antioxidant properties of a radical-scavenging peptide purified from enzymatically prepared fish skin gelatin hydrolysate. J Agric Food Chem 53:581–587

    Article  Google Scholar 

  • Mitsuda H, Yasumoto K, Iwami I (1966) Antioxidative action of indole compounds during the autoxidation of linoleic acid. Eiyoto Shokuryo 19:210–214

    Article  CAS  Google Scholar 

  • Nalinanon S, Benjakul S, Kishimura H, Shahidi F (2001) Functionalities and antioxidant properties of protein hydrolysates from the muscle of ornate threadfin bream treated with pepsin from skipjack tuna. Food Chem 124:1354–1362

    Article  Google Scholar 

  • Nielsen PM, Petersen D, Dambmann C (2001) Improved method for determining food protein degree of hydrolysis. J Food Sci 66:642–646

    Article  CAS  Google Scholar 

  • Palashoff MH (2008) Determining the specificity of pepsin for proteolytic digestion. Department of Chemistry and Chemical Biology, Northeastern University, Boston, MS thesis, p. 78

  • Pham-Huy LA, He H, Pham-Huy C (2008) Free radicals, antioxidants in disease and health. Inter J Biomed Sci 4:89–96

    CAS  Google Scholar 

  • Picot L, Bordenave S, Didelot S, Fruitier-Arnaudin I, Sannier F, Thorkelsson G et al (2006) Antiproliferative activity of fish protein hydrolysates on human breast cancer cell lines. Process Biochem 41:1217–1222

    Article  CAS  Google Scholar 

  • Qian ZJ, Jung WK, Byun HG, Kim SK (2008) Protective effect of an antioxidative peptide purified from gastrointestinal digests of oyster. Crassostrea gigas against free radical induced DNA damage. Bioresour Technol 99:3365–3371

    Article  CAS  PubMed  Google Scholar 

  • Raghavan S, Kristinsson HG, Leeuwenburgh C (2008) Radical scavenging and reducing ability of tilapia (Oreochromis niloticus) protein hydrolysates. J Agric Food Chem 56:10359–10367

    Article  CAS  PubMed  Google Scholar 

  • Rajapakse N, Mendis E, Byun HG, Kim SK (2005) Purification and in vitro antioxidative effects of giant squid muscle peptides on free radical-mediated oxidative systems. J Nutr Biochem 16:562–569

    Article  CAS  PubMed  Google Scholar 

  • Ramarathnam N, Osawa T, Namiki M, Kawakishi S (1988) Chemical studies on novel rice hull antioxidants. I. Isolation, fractionation partial characterization. J Agric Food Chem 36:732–737

    Article  CAS  Google Scholar 

  • Sastry CSP, Tammuru MK (1985) Spectrophotometric determination of tryptophan in proteins. J Food Sci Tech 22:146–147

    CAS  Google Scholar 

  • Sathivel S, Smiley S, Prinyawiwatkul W, Bechtel PJ (2005) Functional and nutritional properties of red salmon (Oncorhynchus nerka) enzymatic hydrolysates. J Food Sci 70:401–406

    Article  Google Scholar 

  • Shahidi F, Zhong Y (2008) Bioactive peptides. J AOAC Int 91:914–931

    CAS  PubMed  Google Scholar 

  • Shan L, Wang B, Gao G, Cao W, Zhang Y (2013) l-Arginine supplementation improves antioxidant defenses through l-arginine/nitric oxide pathways in exercised rats. J Appl Physiol 115:1146–1155

    Article  CAS  PubMed  Google Scholar 

  • Sharp JS, Becker JM, Hettich RL (2003) Protein surface mapping by chemical oxidation: structural analysis by mass spectrometry. Anal Biochem 313:216–225

    Article  CAS  PubMed  Google Scholar 

  • Singleton VL, Rossi JA Jr (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

    CAS  Google Scholar 

  • Souissi N, Bougatef A, Triki-Ellouz Y, Nasri M (2007) Biochemical and functional properties of Sardinella (Sardinella aurita) by-product hydrolysates. Food Technol Biotechnol 45:187–194

    CAS  Google Scholar 

  • Stawikowski M, Fields GB (2002) Introduction to peptide synthesis. In: Coligan JE et al (eds) Current protocols in protein science. Wiley, New York

    Google Scholar 

  • Tanaka H, Fukahori S, Baba S, Ueno T, Sivakumar R, Yagi M, Asagiri K, Ishii S, Tanaka Y (2014) Branched-chain amino acid-rich supplements containing microelements have antioxidant effects on nonalcoholic steatohepatitis in mice. J Parenter Enteral Nutr

  • Udenigwe CC, Aluko RE (2011) Chemometric analysis of the amino acid requirements of antioxidant food protein hydrolysates. Int J Molecular Sci 12:3148–3161

    Article  CAS  Google Scholar 

  • von Gadow A, Joubert E, Hansmann CF (1997) Comparison of the antioxidant activity of aspalathin with that of other plant phenols of rooibos tea (Aspalathus linearis), alpha-tocopherol, BHT, and BHA. J Agric Food Chem 45:632–638

    Article  Google Scholar 

  • Wallner S, Hermetter A, Mayer B, Wascher TC (2001) The alpha-amino group of l-arginine mediates its antioxidant effect. Eur J Clin Invest 31:98–102

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Zhu F, Han F, Wang H (2008) Purification and characterization of antioxidative peptides from salmon protamine hydrolysate. J Food Biochem 32:654–671

    Article  CAS  Google Scholar 

  • Wu HC, Chen HM, Shiau CY (2003) Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). Food Res Int 36:949–957

    Article  CAS  Google Scholar 

  • Zhang Z, Smith DL (1993) Determination of amide hydrogen exchange by mass spectrometry: a new tool for protein structure elucidation. Protein Sci 2:522–553

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhu K, Zhou H, Qian H (2006) Antioxidant and free radical scavenging activities of wheat germ protein hydrolysates WGPH prepared with Alcalase. Process Biochem 41:1296–1302

    Article  CAS  Google Scholar 

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Acknowledgment

Funding

This study was funded by Indian Council of Agricultural research (ICAR), New Delhi.

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Authors and Affiliations

  1. Biochemistry and Nutrition Division, Central Institute of Fisheries Technology, Cochin, 682029, India

    K. K. Asha, K. R. Remya Kumari, K. Ashok Kumar, Niladri S. Chatterjee, R. Anandan & Suseela Mathew

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  1. K. K. Asha
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  2. K. R. Remya Kumari
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  3. K. Ashok Kumar
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  4. Niladri S. Chatterjee
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  5. R. Anandan
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  6. Suseela Mathew
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Correspondence to K. K. Asha.

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Each of the authors, namely, Asha K. K., Remyakumari K. Raghavan, Ashok Kumar K., Niladri S. Chatterjee, R. Anandan and Seseela Mathew declare that he/she has no conflict of interest.

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This article does not contain any studies with human participants performed by any of the authors. This article also does not contain any studies with animals performed by any of the authors.

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Asha, K.K., Remya Kumari, K.R., Ashok Kumar, K. et al. Sequence Determination of an Antioxidant Peptide Obtained by Enzymatic Hydrolysis of Oyster Crassostrea madrasensis (Preston). Int J Pept Res Ther 22, 421–433 (2016). https://doi.org/10.1007/s10989-016-9521-0

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