European Food Research and Technology

, Volume 229, Issue 5, pp 795–805 | Cite as

An approach to improve ACE-inhibitory activity of casein hydrolysates with plastein reaction catalyzed by Alcalase

Original Paper

Abstract

The preparation method of casein hydrolysates with high ACE-inhibitory activity was studied by Alcalase-catalyzed hydrolysis coupled with plastein reaction. Casein hydrolysates with an IC50 value of about 47 μg mL−1 were first prepared by hydrolysis of casein with Alcalase and then modified with plastein reaction catalyzed by the same enzyme. The impacts of four reaction conditions on plastein reaction of casein hydrolysates were studied, and then optimal conditions were determined using response surface methodology with the decrease of free amino groups in the reaction mixture as response. When the concentration of casein hydrolysates was fixed at 35% by weight, the maximum decrease of free amino groups in the reaction mixture of 181.8 μmol g−1 proteins was obtained. The optimum conditions for the above decrease were found to be an E/S ratio of 7.7 kU g−1 proteins, reaction temperature of 42.7 °C and reaction time of 6 h. Analysis results showed that ACE-inhibitory activity of casein hydrolysates prepared could be improved significantly by plastein reaction. When casein hydrolysates were modified by plastein reaction, with a decrease of free amino groups in the mixture of about 154.7 μmol g−1 proteins and 181.8 μmol g−1 proteins, their IC50 values could be decreased to 0.6 and 0.5 μg mL−1.

Keywords

Casein Hydrolysis ACE-inhibitory activity Plastein reaction Alcalase 

References

  1. 1.
    Andrews AT, Alichanidis E (1990) The plastein reaction revisited: evidence for a purely aggregation reaction mechanism. Food Chem 35(4):243–261CrossRefGoogle Scholar
  2. 2.
    Ashley DVM, Temler R, Barclay D, Dormond CA, Jost R (1983) Amino acid-enriched plasteins: a source of limiting amino acids for the weanling rat. J Nutr 113(1):21–27Google Scholar
  3. 3.
    Cheung HS, Wang FL, Ondetti MA, Sabo EF, Cushman DW (1980) Binding of peptide substrates and inhibitors of the angiotensin-converting enzyme. J Biol Chem 255(25):401–407Google Scholar
  4. 4.
    Church FC, Swaisgood HE, Porter DH, Catignani GL (1983) Spectrophotometric assay using ο-Phthaldialdehyde for determination of proteolysis in milk and milk proteins. J Dairy Sc 66(6):1219–1277Google Scholar
  5. 5.
    Combes D, Lozano P (1993) α-Chymotrypsin in plastein synthesis. Influence of water activity. Annals of the New York Academy of Sciences 672 (Enzyme Engineering XI), pp 409–414Google Scholar
  6. 6.
    da Costa EL, da Rocha Gontijo JA, Netto FM (2007) Effect of heat and enzymatic treatment on the antihypertensive activity of whey protein hydrolysates. Int Dairy J 17(6):632–640CrossRefGoogle Scholar
  7. 7.
    Doucet D, Gauthier SF, Otter DE, Foegeding EA (2003) Enzyme-induced gelation of extensively hydrolyzed whey proteins by Alcalase: comparison with the plastein reaction and characterization of interactions. J Agric Food Chem 51(20):6036–6042CrossRefGoogle Scholar
  8. 8.
    FitzGerald RJ, Murray BA, Walsh DJ (2004) Hypotensive peptides from milk proteins. J Nutr 134(4):980S–988SGoogle Scholar
  9. 9.
    Guo Y, Pan D, Tanokura M (2009) Optimisation of hydrolysis conditions for the production of the angiotensin-I-converting enzyme (ACE) inhibitory peptides from whey protein using response surface methodology. Food Chem 114(1):328–333CrossRefGoogle Scholar
  10. 10.
    Haque E, Chand R (2008) Antihypertensive and antimicrobial bioactive peptides from milk proteins. Eur Food Res Technol 227(1):7–15CrossRefGoogle Scholar
  11. 11.
    IDF (1993) Determination of the nitrogen (Kjeldahl method) and calculation of the crude protein content. International Dairy Federation IDF Standard 20B, Brussels, BelgiumGoogle Scholar
  12. 12.
    Jiang J, Chen S, Ren F, Luo Z, Zeng SS (2007) Yak milk casein as a functional ingredient: preparation and identification of angiotensin-I-converting enzyme inhibitory peptides. J Dairy Res 74(1):18–25CrossRefGoogle Scholar
  13. 13.
    Korhonen H, Pihlanto A (2006) Bioactive peptides: production and functionality. Int Dairy J 16(9):945–960CrossRefGoogle Scholar
  14. 14.
    Li G-H, Le G-W, Shi Y-H, Shrestha S (2004) Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects. Nutr Res 24(7):469–486Google Scholar
  15. 15.
    Li GH, Shi YH, Liu H, Le GW (2006) Antihypertensive effect of Alcalase-generated mung bean protein hydrolysates in spontaneously hypertensive rats. Eur Food Res Technol 222(5–6):733–736CrossRefGoogle Scholar
  16. 16.
    López-Fandiño R, Otte J, van Camp J (2006) Physiological, chemical and technological aspects of milk-protein-derived peptides with antihypertensive and ACE-inhibitory activity. Int Dairy J 16(11):1277–1293CrossRefGoogle Scholar
  17. 17.
    Lozano P, Combes D (1991) α-Chymotrypsin in plastein synthesis: influence of substrate concentration on enzyme activity. Biotechnol Appl Biochem 14(2):212–221Google Scholar
  18. 18.
    Mao XY, Ni JR, Sun WL, Hao PP, Fan L (2007) Value-added utilization of yak milk casein for the production of angiotensin-I-converting enzyme inhibitory peptides. Food Chem 103(4):1282–1287CrossRefGoogle Scholar
  19. 19.
    Megías C, Yust MM, Pedroche J, Lquari H, Girón-Calle J, Alaiz M, Millán F, Vioque J (2004) Purification of an ACE-inhibitory peptide after hydrolysis of sunflower (Helianthus annuus L.) protein isolates. J Agric Food Chem 52(7):1928–1932CrossRefGoogle Scholar
  20. 20.
    Miguel M, Alonso MJ, Salaices M, Aleixandre A, López-Fandiño R (2007) Antihypertensive, ACE-inhibitory and vasodilator properties of an egg white hydrolysate: effect of a simulated intestinal digestion. Food Chem 104(1):163–168CrossRefGoogle Scholar
  21. 21.
    Miguel M, Contreras MM, Recio I, Aleixandre A (2009) ACE-inhibitory and antihypertensive properties of a bovine casein hydrolysate. Food Chem 112(1):211–214CrossRefGoogle Scholar
  22. 22.
    Minervini F, Algaron F, Rizzello CG, Fox PF, Monnet V, Gobbetti M (2003) Angiotensin I-converting-enzyme-inhibitory and antibacterial peptides from Lactobacillus helveticus PR4 proteinase-hydrolyzed caseins of milk from six species. Appl Environ Microbiol 69(9):5297–5305CrossRefGoogle Scholar
  23. 23.
    Murray BA, Walsh DJ, FitzGerald RJ (2004) Modification of the furanacryloyl- l-phenylalanyl-glycylglycine assay for determination of angiotensin-I-converting enzyme inhibitory activity. J Biochem Biophys Methods 59(2):127–137CrossRefGoogle Scholar
  24. 24.
    Nakamura Y, Yamamoto N, Sakai K, Okubo A, Yamazaki S, Takano T (1995) Purification and characterization of angiotensin-I converting enzyme inhibitors from a sour milk. J Dairy Sci 78(4):777–783CrossRefGoogle Scholar
  25. 25.
    Ortiz-Chao P, Gómez-Ruiz JA, Rastall RA, Mills D, Cramer R, Pihlanto A, Korhonen H, Jauregi P (2009) Production of novel ACE-inhibitory peptides from β-lactoglobulin using Protease N Amano. Int Dairy J 19(2):69–76CrossRefGoogle Scholar
  26. 26.
    Otte J, Shalaby SM, Zakora M, Pripp AH, El-Shabrawy SA (2007) Angiotensin-converting enzyme inhibitory activity of milk protein hydrolysates: effect of substrate, enzyme and time of hydrolysis. Int Dairy J 17(5):488–503CrossRefGoogle Scholar
  27. 27.
    Otte J, Shalaby SMA, Zakora M, Nielsen MS (2007) Fractionation and identification of ACE-inhibitory peptides from α-lactalbumin and β-casein produced by thermolysin-catalysed hydrolysis. Int Dairy J 17(12):1460–1472CrossRefGoogle Scholar
  28. 28.
    Pallavicini C, Finley JW, Stanley WL, Watters GG (1980) Plastein synthesis with α-chymotrypsin immobilised on chitin. J Sci Food Agric 31(3):273–278CrossRefGoogle Scholar
  29. 29.
    Pihlanto A, Akkanen S, Korhonen HJ (2008) ACE-inhibitory and antioxidant properties of potato (Solanum tuberosum). Food Chem 109(1):104–112CrossRefGoogle Scholar
  30. 30.
    Pihlanto-Leppälä A (2000) Bioactive peptides derived from bovine whey proteins: opioid and ace-inhibitory peptides. Trends Food Sci Technol 11(9–10):347–356CrossRefGoogle Scholar
  31. 31.
    Robert MC, Razaname A, Mutter M, Juillerat MA (2004) Identification of angiotensin-I-converting enzyme inhibitory peptides derived from sodium caseinate hydrolysates produced by Lactobacillus helveticus NCC 2765. J Agric Food Chem 52(23):6923–6931CrossRefGoogle Scholar
  32. 32.
    Shalaby SM, Zakora M, Otte J (2006) Performance of two commonly used angiotensin-converting enzyme inhibition assays using FA-PGG and HHL as substrates. J Dairy Res 73(2):178–186CrossRefGoogle Scholar
  33. 33.
    Spellman D, McEvoy E, O’Cuinn G, FitzGerald RJ (2003) Proteinase and exopeptidase hydrolysis of whey protein: comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. Int Dairy J 13(6):447–453CrossRefGoogle Scholar
  34. 34.
    Stevenson DE, Morgan KR, Fenton GA, Moraes G (1999) Use of NMR and mass spectrometry to detect and quantify protease-catalyzed peptide bond formation in complex mixtures. Enzyme Microb Technol 25(3–5):357–363CrossRefGoogle Scholar
  35. 35.
    Stevenson DE, Ofman DJ, Morgan KR, Stanley RA (1998) Protease-catalyzed condensation of peptides as a potential means to reduce the bitter taste of hydrophobic peptides found in protein hydrolysates. Enzyme Microb Technol 22(2):100–110CrossRefGoogle Scholar
  36. 36.
    Suh HJ, Kim KM, Noh DO, Yu KW, Ahn SW (2000) Isolation of angiotensin I converting enzyme inhibitory peptide from soybean hydrolysate. Food Sci Biotechnol 9(6):378–381Google Scholar
  37. 37.
    Sukan G, Andrews AT (1982a) Application of the plastein reaction to caseins and to skim milk powder I. Protein hydrolysis and plastein formation. J Dairy Res 49:265–278CrossRefGoogle Scholar
  38. 38.
    Sukan G, Andrews AT (1982b) Application of the plastein reaction to caseins and to skim milk powder II. Chemical and physical properties of the plasteins. J Dairy Res 49:279–293CrossRefGoogle Scholar
  39. 39.
    Williams RJH, Brownsell VL, Andrews AT (2001) Application of the plastein reaction to mycoprotein: I. Plastein synthesis. Food Chem 72(3):329–335CrossRefGoogle Scholar
  40. 40.
    Yamashita M, Arai S, Fujimaki M (1976a) Plastein reaction for food protein improvement. J Agric Food Chem 24(6):1100–1104CrossRefGoogle Scholar
  41. 41.
    Yamashita M, Arai S, Fujimaki M (1976b) A low-phenylalenine, high-tyrosine plastein as an acceptable dietetic food. J Food Sci 41(5):1029–1032CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Key Laboratory of Dairy Science, Ministry of EducationNortheast Agricultural UniversityHarbinPeople’s Republic of China

Personalised recommendations