Amino Acids

, Volume 43, Issue 1, pp 337–345 | Cite as

Optimization of enzymatic hydrolysis conditions for the production of antioxidant peptides from muscles of Nemipterus japonicus and Exocoetus volitans using response surface methodology

  • Shabeena Yousuf Naqash
  • R. A. NazeerEmail author
Original Article


In the present study, protein of muscles of commercially important marine fishes Nemipterus japonicus and Exocoetus volitans were extracted by trypsin and their hydrolysis conditions viz., temperature, time, and enzyme to substrate concentration on degree of hydrolysis were studied by response surface methodology. The optimum values for N. japonicus was found as temperature, 30°C, hydrolysis time of 100 min an enzyme/substrate concentration of 1.59% whereas, for E. volitans muscle protein, optimum hydrolysis conditions were temperature, 30°C, hydrolysis time of 115 min and enzyme/substrate concentration of 1.67%. Furthermore, amino acid sequence of antioxidant peptides derived after chromatographic purification was identified by ESI–MS/MS. The analysis of peptides showed sequences as Glu-Ser-Asp-Arg-Pro (620.3 Da) and Gly-Trp-Met-Gly-Cys-Trp (747.3) for N. japonicus and E. volitans muscle, respectively. The peptides contained important antioxidant amino acids and acted as good antioxidant peptides to scavenge free radicals.


Nemipterus japonicus Exocoetus volitans Antioxidant peptides Amino acids 



We gratefully acknowledge the management of SRM University for providing the facilities to carry out this project.

Conflict of interest

No competing financial interests exist.


  1. Alder-Nissen J (1986) Enzymic hydrolysis of food proteins. Elsevier Applied Science Publisher, New YorkGoogle Scholar
  2. Ames BN (1983) Dietary carcinogens and anticarcinogens: oxygen radicals and degenerative disease. Science 221:1256–1264PubMedCrossRefGoogle Scholar
  3. Amiza MA, Nurul Ashikin S, Faazaz AL (2011) Optimization of enzymatic protein hydrolysis from silver catfish (Pangasius sp.) frame. Intern Food Res J 18:751–757Google Scholar
  4. Bhaskar N, Benila T, Radha C, Lalitha RG (2007) Optimization of enzymatic hydrolysis of visceral waste proteins of Catla (Catla catla) for preparing protein hydrolysate using commercial protease. Bioresour Technol 99:335–343PubMedCrossRefGoogle Scholar
  5. Erdmann K, Cheung WY, Schröder H (2008) The possible roles of food derived bioactive peptides in reducing the risk of cardiovascular diseases. J Nutr Biochem 19:643–654PubMedCrossRefGoogle Scholar
  6. Giovanni M (1983) Response surface methodology and product optimization. Food Technol 80:41–45Google Scholar
  7. Gulçin I (2009) Antioxidant activity of l-adrenaline: a structure-activity insight. Chem Biol Interact 179:71–80PubMedCrossRefGoogle Scholar
  8. Guo Q, Zhao B, Shen S, Hou J, Hu J, Xin W (1999) ESR study on the structure-antioxidant activity relationship of tea catechins and their epimers. Biochem Biophys Acta 1427:13–23PubMedCrossRefGoogle Scholar
  9. Hernandez-Ledesma B, Davalos A, Bartolome B, Amigo L (2005) Preparation of antioxidant enzymatic hydrolysates from α-lactalbumin and β-lactoglobulin, identification of active peptides by HPLC–MS/MS. J Agric Food Chem 53:588–593PubMedCrossRefGoogle Scholar
  10. Hook VYH, Burton D, Yasothornsrikul S, Hastings RH, Deftos LJ (2001) Proteolysis of ProPTHrP (1–141) by “Prohormone Thiol Protease” at multibasic residues generates PTHrP-related peptides: implications for PTHrP peptide production in lung cancer cells. Biochem Biophys Res Commun 285:932–938PubMedCrossRefGoogle Scholar
  11. Kaur C, Kapoor HC (2001) Antioxidant in fruits and vegetables—the millennium’s health. Int J Food Sci Technol 36:703–725CrossRefGoogle Scholar
  12. Li B, Chen F, Wang X, Ji BP, Wu Y (2007) Isolation and identification of antioxidative peptides from porcine collagen hydrolysate by consecutive chromatography and electrospray ionization mass spectrometry. Food Chem 102:1135–1143CrossRefGoogle Scholar
  13. Liaset B, Lied E, Espe M (2000) Enzymatic hydrolysis of by-products from the fish-filleting industry, chemical characterization and nutritional evaluation. J Sci Food Agric 80:581–589CrossRefGoogle Scholar
  14. Lin CC, Liang JH (2002) Effect of antioxidants on the oxidative stability of chicken breast meat in a dispersion system. J Food Sci 67:530–533CrossRefGoogle Scholar
  15. Madamba PS (2002) The response surface methodology: an application to optimize operations of selected agricultural crops. Food Sci Technol 35:584–592Google Scholar
  16. Nanjo F, Goto K, Seto R, Suzuki M, Sakai M, Hara Y (1996) Scavenging effects of tea catechins and their derivatives on 1,1,-diphenyl-2-picrylhydrazyl radical. Free Radic Biol Med 21:895–902PubMedCrossRefGoogle Scholar
  17. Pihlanto-Leppala A (2000) Bioactive peptides derived from bovine whey proteins: opioid and ACE-inhibitory peptides. Trends Food Sci Technol 11:347–356CrossRefGoogle Scholar
  18. 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 9:562–569CrossRefGoogle Scholar
  19. Ren J, Zhao M, Shi J, Wang J, Jiang Y, Cui C, Kakuda Y, Xue SJ (2008) Optimization of antioxidant peptide production from grass carp sarcoplasmic protein using response surface methodology. Food Sci Technol 41:1624–1632Google Scholar
  20. Rosen GM, Rauckman EJ (1984) Spin trapping of superoxide and hydroxyl radicals. Method Enzymol 105:198–209CrossRefGoogle Scholar
  21. Shabeena YN, Nazeer RA (2010) Antioxidant activity of hydrolysates and peptide fractions of Nemipterus japonicus and Exocoetus volitans muscle. J Aquat Food Prod Technol 19:180–192CrossRefGoogle Scholar
  22. Suetsuna K, Ukeda H, Ochi H (2000) Isolation and characterization of free radical scavenging activity peptides derived from casein. J Nutr Biochem 11:128–131PubMedCrossRefGoogle Scholar
  23. Tsuge N, Eikawa Y, Nomura Y, Yamamoto M, Sugisawa K (1991) Antioxidant activity of peptides prepared by enzymatic hydrolysis of egg-white albumin. Nippon Nogeikagaku Kaishi—J Jpn Soc Biosci Biotechnol Agrochem 65:1635–1641CrossRefGoogle Scholar
  24. Vercruysse L, Van Camp J, Smagghie G (2005) ACE inhibitory peptides derived from enzymatic hydrolysates of animal muscle protein: a review. J Agric Food Chem 53:8106–8115PubMedCrossRefGoogle Scholar
  25. Viera GHF, Martin AM, Sampaiao SS, Omar S, Gonsalves RCF (1995) Studies on the enzymatic hydrolysis of Brazilian lobster (Panulirus spp.) processing wastes. J Sci Food Agric 69:61–65CrossRefGoogle Scholar
  26. Wasswa J, Tang J, Xiao HG (2008) Optimization of the production of hydrolysates from Grass carp (Ctenopharyngodon idella) skin using Alcalase®. J Food Biochem 32:460–473CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Biotechnology, School of BioengineeringSRM UniversityChennaiIndia

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