Advertisement

Purification and Characterization of Antioxidant Peptides from Oyster (Saccostrea cucullata) Hydrolysate and the Anticancer Activity of Hydrolysate on Human Colon Cancer Cell Lines

  • S. Umayaparvathi
  • M. Arumugam
  • S. Meenakshi
  • Gerald Dräger
  • Andreas Kirschning
  • T. Balasubramanian
Article

Abstract

The focus of the study was to investigate antioxidant activity and characterize antioxidant peptides from oyster (Saccostrea cucullata) protein hydrolysate. The protease hydrolysate of oyster exhibited strong potential to donate hydrogen and was able to scavenge Hydrogen peroxide, Hydroxyl and DPPH radicals. Due to the high antioxidant potential, hydrolysate was purified in Sephadex G-25 gel filtration chromatography. The active peptide fraction was further purified by UPLC-MS. Totally seven antioxidant peptides were collected. Among seven peptides (SCAP 1–7), three peptides (SCAP 1, 3 and 7) had highest scavenging ability on DPPH radicals. The amino acid sequence and molecular mass of purified antioxidant peptides (SCAP1, SCAP3 and SCAP7) were determined by Q-TOF ESI mass spectroscopy and structures of the peptides were Leu-Ala-Asn-Ala-Lys (MW = 515.29 Da), Pro-Ser-Leu-Val-Gly-Arg-Pro–Pro-Val-Gly-Lys-Leu-Thr-Leu (MW = 1,432.89 Da) and Val-Lys-Val-Leu-Leu-Glu-His-Pro-Val-Leu (MW = 1,145.75 Da), respectively. The oyster hydrolysate was tested for cell cytotoxicity on Vero (kidney epithelial cells of the African Green Monkey) and HT-29 (human colon carcinoma) cell lines. It was found that the hydrolysate did not show any cytotoxic effect for Vero cell lines and exerted a significant cytotoxic effect on HT-29 cell lines. We thus conclude that the anticancer and antioxidative hydrolysate from oyster (S. cucullata) may be useful ingredients in food and nutraceutical applications.

Keywords

Oyster Saccostrea cucullata Enzymatic hydrolysis Protein hydrolysate Antioxidant peptide DPPH Electrospray ionization mass spectrometry 

Notes

Acknowledgments

The authors would like to thank Ministry Of Earth Sciences for financial support, under the “Drugs from sea” programme and the authors are grateful to the authorities of Annamalai University for providing the necessary facilities.

Conflict of interest

S. Umayaparvathi, M. Arumugam, S. Meenakshi, G. Draeger, A. Kirschning and T. Balasubramanian declare that they have no conflict of interest.

Statement of informed consent/Human and animal rights

This article does not contain any studies with human or animal subjects performed by the any of the authors.

References

  1. Aleman A, Perez-Santin E, Bordenave-Juchereau S, Arnaudin I, Gomez-Guillen MC, Montero P (2011) Squid gelatin hydrolysates with antihypertensive, anticancer and antioxidant activity. Food Res Int 44:1044–1051CrossRefGoogle Scholar
  2. AOAC (1990) Official methods of analysis. Association of Official Analytical Chemists, Washington, DCGoogle Scholar
  3. Blunt JW, Copp BR, Munro MHG, Northcote PT, Prinsep MR, Rep NP (2005) Marine natural products. Nat prod Rep 22:15–61PubMedCrossRefGoogle Scholar
  4. 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 114:1198–1205CrossRefGoogle Scholar
  5. 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 118:559–565CrossRefGoogle Scholar
  6. Byun HG, Lee JK, Park HG, Jeon JK, Kim SK (2009) Antioxidant peptides isolated from the marine rotifer, Brachionus rotundiformis. Process Biochem 44:842–846CrossRefGoogle Scholar
  7. Chen HM, Muramoto K, Yamaguchi F (1995a) Structural analysis of antioxidative peptides from soybean beta-conglycinin. J Agric Food Chem 43(3):574–578CrossRefGoogle Scholar
  8. Chen J, Suetsuna K, Yamauchi F (1995b) Isolation and characterization of immunostimulative peptides from soybean. J Nutr Biochem 6:310–313CrossRefGoogle Scholar
  9. Chen HM, Muramoto K, Yamauchi F, Nokihara K (1996) Antioxidant activity of designed peptides based on the antioxidative peptide isolated from digests of a soybean protein. J Agric Food Chem 44:2619–2623CrossRefGoogle Scholar
  10. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric methods for determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  11. Folch J, Lees M, Solam-Stanley GH (1957) A simple method for the isolation and purification of claot lipid from animal tissue. J Biol Chem 226:497–509PubMedGoogle Scholar
  12. Halliwell B, Gutteridge JMC (1990) Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol 186:1–85PubMedCrossRefGoogle Scholar
  13. Hancock JT, Desikan R, Neil SJ (2001) Role of reactive oxygen species in cell signaling pathways. Biochem Soc Trans 29:345–350PubMedCrossRefGoogle Scholar
  14. He H, Chen X, Sun C, Zhang Y, Gao P (2006) Preparation and functional evaluation of oligopeptide-enriched hydrolysate from shrimp (Acetes chinensis) treated with crude protease from Bacillus sp. SM98011. Bioresour Technol 97:385–390PubMedCrossRefGoogle Scholar
  15. Hettiarachchy NS, Glenn KC, Gnanasambandam R, Johnson MG (1996) Natural antioxidant extract from fenugreek (Trigonella foenumgraecum) for ground beef patties. J Food Sci 61:516–519CrossRefGoogle Scholar
  16. Huang DC, Li QF, Li P, Li XQ, Song YZ (2002) Effects of oyster low molecular weight bioactive substance on the human lung adenocarcinoma A549 cells. J Xiamen Univ 41:614–617Google Scholar
  17. Huang F, Yang Z, Yu D, Wang J, Li R, Ding G (2012) Sepia Ink oligopeptide induces apoptosis in prostate cancer cell lines via caspase-3 activation and elevation of Bax/Bcl-2 ratio. Mar Drugs 10:2153–2165PubMedCentralPubMedCrossRefGoogle Scholar
  18. Hunt DA, Miescier J, Redman J, Salinger A, Lucas JP (1984) Molluscan shellfish, fresh or fresh frozen oysters, mussels or clams. In: Speck ML (ed) Compendium of methods for the microbiological examination of foods, 2nd edn. American Public Health Association, Washington, DCGoogle Scholar
  19. Jumeri, Kim SM (2011) Antioxidant and anticancer activities of enzymatic hydrolysates of solitary tunicate (Styela clava). Food Sci Biotechnol 20(4):1075–1085CrossRefGoogle Scholar
  20. Kaufman SH, Earnshaw WC (2000) Induction of apoptosis by cancer chemotherapy. Cell Res 256:42–49CrossRefGoogle Scholar
  21. Kembhavi AA, Kulkarni A, Pant A (1993) Salt-tolerant and thermostable alkaline protease from Bacillus subtilis NCIM No. 64. Appl Biochem Biotechnol 38(8):3–92Google Scholar
  22. Leanderson P, Faresjo AO, Tagesson C (1997) Green tea polyphenols inhibits oxidant-induced DNA strand breakage in cultured lung cells. Free Radical Biol Med 23:235–242CrossRefGoogle Scholar
  23. Leng B, Liu XD, Chen QX (2005) Inhibitory effects of anticancer peptide from Mercenaria on the BGC-823 cells and several enzymes. FEBS 579:1187–1190CrossRefGoogle Scholar
  24. Lind DS, Hochwald SN, Malaty J, Rekkas S, Hebig P, Mishra G et al (2001) Nuclear factor-kB is upregulated in colorectal cancer. Surgery 130:363–369PubMedCrossRefGoogle Scholar
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  26. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63PubMedCrossRefGoogle Scholar
  27. Oyaizu M (1986) Studies on products of browning reaction: antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 44:307–315CrossRefGoogle Scholar
  28. Pihlanto-Leppala A (2001) Bioactive peptides derived from whey proteins: opioid and ACE inhibitory peptides. Trends Food Sci Technol 11:347–356CrossRefGoogle Scholar
  29. Prieto P, Pineda M, Aguilar M (1999) Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 269:337–341PubMedCrossRefGoogle Scholar
  30. 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 99:3365–3371PubMedCrossRefGoogle Scholar
  31. Ruch RJ, Cheng SJ, Klauning JE (1989) Prevention of cytotoxity and inhibition of intercellular communication by antioxidant catechin isolated from Chinese green tea. Carcinogenesis 10:1003–1008PubMedCrossRefGoogle Scholar
  32. Rustad T (2003) Utilisation of marine by-products. Electron J Environ Agric Food Chem 2:458–463Google Scholar
  33. Sakanaka S, Tachibana Y (2006) Active oxygen scavenging activity of egg-yolk protein hydrolysates and their effects on lipid oxidation in beef and tuna homogenates. Food Chem 95:243–249CrossRefGoogle Scholar
  34. Seril DN, Liao J, Yang GY, Yang CS (2003) Oxidative stress and ulcerative colitis asso-ciated carcinogenesis: studies in humans and animal models. Carcinogenesis 24:353–362PubMedCrossRefGoogle Scholar
  35. Sikorski ZE, Naczk M (1981) Modification of technological properties of fish protein concentrate. CRC Crit Rev Food Sci Nutr 14:201–230CrossRefGoogle Scholar
  36. Simpson BK, Nayeri G, Yaylayan V, Ashie INA (1998) Enzymatic hydrolysis of shrimp meat. Food Chem 61:131–138CrossRefGoogle Scholar
  37. Song L, Li T, Yu R, Yan C, Ren S, Zhao Y (2008a) Antioxidant activities of hydrolysates of Arca Subcrenata prepared with three proteases. Mar Drugs 6:607–619PubMedCentralPubMedCrossRefGoogle Scholar
  38. Song L, Ren S, Yu R, Yan C, Li T, Zhao Y (2008b) Purification, characterization and in vitro anti-tumor activity of proteins from Arca subcrenata lischke. Mar Drugs 6:418–430PubMedCentralPubMedCrossRefGoogle Scholar
  39. Wang Y, Ma AL, Zhang HZ, Xue BH, Zhao ZJ, Fu FH, Zhou GY (1997) Experimental studies on the antitumor effect of oyster extract. Chin J Mar Drug 16:18–22Google Scholar
  40. 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 36:949–957CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • S. Umayaparvathi
    • 1
  • M. Arumugam
    • 1
  • S. Meenakshi
    • 1
  • Gerald Dräger
    • 2
  • Andreas Kirschning
    • 2
  • T. Balasubramanian
    • 1
  1. 1.Centre of Advanced Study in Marine Biology, Faculty of Marine SciencesAnnamalai UniversityParangipettaiIndia
  2. 2.Institute of Organic ChemistryUniversity of HannoverHannoverGermany

Personalised recommendations