Journal of Food Science and Technology

, Volume 51, Issue 9, pp 1847–1856 | Cite as

ACE inhibitory activity of pangasius catfish (Pangasius sutchi) skin and bone gelatin hydrolysate

  • Fatemeh Mahmoodani
  • Masomeh Ghassem
  • Abdul Salam Babji
  • Salma Mohamad Yusop
  • Roya Khosrokhavar
Original Article


Skin and bone gelatins of pangasius catfish (Pangasius sutchi) were hydrolyzed with alcalase to isolate Angiotensin Converting Enzyme (ACE) inhibitory peptides. Samples with the highest degree of hydrolysis (DH) were separated into different fractions with molecular weight cut-off (MWCO) sizes of 10, 3 and 1 kDa, respectively and assayed for ACE inhibitory activity. Skin and bone gelatins had highest DH of 64.87 and 68.48 % after 2 and 1 h incubation, respectively. Results from this study indicated that by decreasing the molecular weight of fractions, ACE inhibitory activity was increased. Therefore, F3 permeates (MWCO < 1 kDa) of skin (IC50 = 3.2 μg/ml) and bone (IC50 = 1.3 μg/ml) gelatins possessed higher ACE inhibitory activity compared to their untreated gelatins and corresponding hydrolyzed fractions. In this study, the major amino acids were Glycine followed by Proline with an increased amount of hydrophobic amino acid content in F3 permeates of skin (4.01 %) and bone (5.79 %) gelatin. Digestion stability against gastrointestinal proteases did not show any remarkable change on ACE inhibition potency of these permeates. It was concluded that alcalase hydrolysis of P. sutchi by-products could be utilized as a part of functional food or ingredients of a formulated drug in order to control high blood pressure.


Enzymatic hydrolysis ACE inhibitory Pangasius sutchi Gelatin High blood pressure 


  1. Alaiz M, Navarro JL, Girón J, Vioque E (1992) Amino acid analysis by high-performance liquid chromatography after derivatization with diethyl ethoxymethylenemalonate. J Chromatogr 591:181–186CrossRefGoogle Scholar
  2. Alemán A, Giménez B, Pérez-Santín E, Gómez-Guillén MC, Montero P (2011) Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptides sequences isolated from squid gelatin hydrolysate. Food Chem 125:334–341CrossRefGoogle Scholar
  3. Alfaro AT, Costa CS, Fonseca GG, Prentice C (2009) Effect of extraction parameters on the properties of gelatin from King weakfish (Macrodon ancylodon) bones. Food Sci Tech Int 15:553–562CrossRefGoogle Scholar
  4. AOAC (2005) Official methods of the association of official agricultural Chemist’s International, 17th edn. Gaithersburg, AOAC InternationalGoogle Scholar
  5. Benjakul S, Morrissey MT (1997) Protein hydrolysates from pacific whiting solid wastes. J Agric Food Chem 45:3423–3430CrossRefGoogle Scholar
  6. Bougatef A, Nedjar-Arroume N, Ravallec-Plé R, Leroy Y, Guillochon D, Barkia A, Nasri M (2008) Angiotensin I-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases. Food Chem 111:350–356CrossRefGoogle Scholar
  7. Byun HG, Kim SK (2001) Purification and characterization of angiotensin I converting enzyme (ACE) inhibitory peptides from Alaska pollack (Theragra chalcogramma) skin. Process Biochem 36:1155–1162CrossRefGoogle Scholar
  8. Chen HM, Muramoto K, Yamauchi F (1995) Structural analysis of antioxidative peptides from soybean β-conglycinin. J Agric Food Chem 43:574–578CrossRefGoogle Scholar
  9. Church FC, Swaisgood HE, Porter DH, Catignani GL (1983) Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. J Dairy Sci 66:1219–1227CrossRefGoogle Scholar
  10. Cohen SA, Meys M, Tarvin TL (1988) The PicoTag Method. A manual of advanced techniques for amino acid analysis. Waters Chromatography Division, Millipore Corp, Milford, MAGoogle Scholar
  11. Diniz FM, Martin AM (1996) Use of response surface methodology to describe the combined effects of pH, temperature and E/S ratio on the hydrolysis of dogfish (Squalus acanthias) muscle. Int J Food Sci Tech 31:419–426CrossRefGoogle Scholar
  12. Fahmi A, Morimura S, Guo HC, Shigematsu T, Kida K, Uemura Y (2004) Production of angiotensin I converting enzyme inhibitory peptides from sea bream scales. Process Biochem 39:1195–1200CrossRefGoogle Scholar
  13. Fujita H, Yokoyama K, Yoshikawa M (2000) Classification and antihypertensive activity of angiotensin I-converting enzyme inhibitory peptides derived from food protein. J Food Sci 65:564–569CrossRefGoogle Scholar
  14. Ghassem M, Siau Fern S, Said M, Mohd Ali Z, Ibrahim S, Salam Babji A (2011a) Kinetic characterization of Channa striatus muscle sarcoplasmic and myofibrillar protein hydrolysates. J Food Sci Technol. doi:10.1007/s13197-011-0526-6
  15. Ghassem M, Arihara K, Babji AS, Said M, Ibrahim S (2011b) Purification and identification of ACE inhibitory peptides from Haruan (Channa striatus) myofibrillar protein hydrolysate using HPLC–ESI-TOF MS/MS. Food Chem 129:1770–1777CrossRefGoogle Scholar
  16. Giménez B, Alemán A, Montero P, Gómez-Guillén MC (2009) Antioxidant and functional properties of gelatin hydrolysates obtained from skin of sole and squid. Food Chem 114:976–983CrossRefGoogle Scholar
  17. Gómez-Guillén MC, Giménez B, López-Caballero ME, Montero MP (2011) Functional and bioactive properties of collagen and gelatin from alternative sources: a review. Food Hydrocoll 25:1813–1827CrossRefGoogle Scholar
  18. Gómez-Ruiz JÁ, Ramos M, Recio I (2004) Identification and formation of angiotensin-converting enzyme-inhibitory peptides in Manchego cheese by high-performance liquid chromatography–tandem mass spectrometry. J Chromatogr A 1054:269–277CrossRefGoogle Scholar
  19. Guérard F, Guimas L, Binet A (2002) Production of tuna waste hydrolysates by a commercial neutral protease preparation. J Mol Catal B: Enzym 19:489–498CrossRefGoogle Scholar
  20. Harnedy PA, Fitzgerald RJ (2012) Bioactive peptides from marine processing waste and shellfish: a review. J Funct Foods 4:6–24CrossRefGoogle Scholar
  21. Hernández-Ledesma B, Del Mar CM, Recio I (2011) Antihypertensive peptides: production, bioavailability and incorporation into foods. Adv Colloid Interface Sci 165:23–35CrossRefGoogle Scholar
  22. Hoyle NT, Merritt JH (1994) Quality of fish protein hydrolysate from Herring (Clupea harengus). J Food Sci 59:76–79CrossRefGoogle Scholar
  23. Ichimura T, Yamanaka A, Otsuka T, Yamashita E, Maruyama S (2009) Antihypertensive effect of enzymatic hydrolysate of collagen and Gly-Pro in spontaneously hypertensive rats. Biosci Biotechnol Biochem 73:2317–2319CrossRefGoogle Scholar
  24. Jeon Y, Byun H, Kim S (1999) Improvement of functional properties of cod frame protein hydrolysates using ultrafiltration membranes. Process Biochem 35:471–478CrossRefGoogle Scholar
  25. Jimsheena VK, Gowda LR (2011) Angiotensin I-converting enzyme (ACE) inhibitory peptides derived from arachin by simulated gastric digestion. Food Chem 125:561–569CrossRefGoogle Scholar
  26. Karim A, Bhat R (2009) Fish gelatin: properties, challenges, and prospects as an alternative to mammalian gelatins. Food Hydrocoll 23:563–576CrossRefGoogle Scholar
  27. Kim SK, Byun HG, Park PJ, Shahidi F (2001) Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. J Agric Food Chem 49:2992–2997CrossRefGoogle Scholar
  28. Kim SY, Je JY, Kim SK (2007) Purification and characterization of antioxidant peptide from hoki (Johnius belengerii) frame protein by gastrointestinal digestion. J Nutr Biochem 18:31–38CrossRefGoogle Scholar
  29. Klompong V, Benjakul S, Kantachote D, Shahidi F (2007) Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (Selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type. Food Chem 102:1317–1327CrossRefGoogle Scholar
  30. Korhonen H, Pihlanto A (2003) Food-derived bioactive peptides-opportunities for designing future foods. Curr Pharm Des 9:1297–1308CrossRefGoogle Scholar
  31. Kristinsson HG, Rasco BA (2000a) Biochemical and functional properties of Atlantic salmon (Salmo salar) muscle hydrolyzed with various alkaline proteases. J Agric Food Chem 48:657–666CrossRefGoogle Scholar
  32. Kristinsson HG, Rasco BA (2000b) Fish protein hydrolysates: production, biochemical and functional properties. Food Sci Nutr 40:43–81Google Scholar
  33. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277:680–685CrossRefGoogle Scholar
  34. Lee JK, Hong S, Jeon JK, Kim SK, Byun HG (2009) Purification and characterization of angiotensin I converting enzyme inhibitory peptides from the rotifer, Brachionus rotundiformis. Bioresour Technol 100:5255–5259CrossRefGoogle Scholar
  35. Lee JK, Jeon JK, Byun HG (2011) Effect of angiotensin I converting enzyme inhibitory peptide purified from skate skin hydrolysate. Food Chem 125:495–499CrossRefGoogle Scholar
  36. Li H, Aluko RE (2010) Identification and inhibitory properties of multifunctional peptides from pea protein hydrolysate. J Agric Food Chem 58:11471–11476CrossRefGoogle Scholar
  37. Lin L, Li BF (2006) Radical scavenging properties of protein hydrolysates from jumbo flying squid (Dosidicus eschrichitii steenstrup) skin gelatin. J Sci Food Agric 86:2290–2295CrossRefGoogle Scholar
  38. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biolog Chem 193:265–275Google Scholar
  39. Majumder K, Wu J (2011) Purification and characterisation of angiotensin I converting enzyme (ACE) inhibitory peptides derived from enzymatic hydrolysate of ovotransferrin. Food Chem 126:1614–1619CrossRefGoogle Scholar
  40. Meisel H (2005) Biochemical properties of peptides encrypted in bovine milk proteins. Curr Med Chem 12:1905–1919CrossRefGoogle Scholar
  41. Mendis E, Rajapakse N, Byun H, Kim S (2005) Investigation of jumbo squid (Dosidicus gigas) skin gelatin peptides for their in vitro antioxidant effects. Life Sci 77:2166–2178CrossRefGoogle Scholar
  42. Montero P, Gómez-Guillén MC (2000) Extracting conditions for megrim (Lepidorhombus boscii) skin collagen affect functional properties of the resulting gelatin. J Food Sci 65:434–438CrossRefGoogle Scholar
  43. Mullally MM, Meisel H, FitzGerald RJ (1997) Identification of novel angiotensin-I converting enzyme inhibitory peptide corresponding to tryptic fragment of bovine beta lactoglobulin. FEBS Lett 402:99–101CrossRefGoogle Scholar
  44. Nagai T, Nagashima T, Abe A, Suzuki N (2006) Antioxidative activities and angiotensin I-converting enzyme inhibition of extracts prepared from chum salmon (Oncorhynchus keta) cartilage and skin. Int J Food Prop 9:813–822CrossRefGoogle Scholar
  45. Nam KA, You SG, Kim SM (2008) Molecular and physical characteristics of squid (Toradores pacificus) skin collagens and biological properties of their enzymatic hydrolysates. J Food Sci 73:249–255CrossRefGoogle Scholar
  46. Pan D, Cao J, Guo H, Zhao B (2012) Studies on purification and the molecular mechanism of a novel ACE inhibitory peptide from whey protein hydrolysate. Food Chem 130:121–126CrossRefGoogle Scholar
  47. Park P, Jung W, Nam K, Shahidi F, Kim S (2001) Purification and characterization of antioxidative peptides from protein hydrolysate of lecithin-free egg yolk. JAOCS, J Am Oil Chem Soc 78:651–656CrossRefGoogle Scholar
  48. Park CH, Kim HJ, Kang KT, Park JW, Kim JS (2009) Fractionation and angiotensin I-converting enzyme (ACE) inhibitory activity of gelatin hydrolysates from by-products of Alaska pollock surimi. Fish Aquat Sci 12:79–85Google Scholar
  49. Phelan M, Aherne A, Fitzgerald RJ, O’Brien NM (2009) Casein-derived bioactive peptides: biological effects, industrial uses, safety aspects and regulatory status. Int Dairy J 19:643–654CrossRefGoogle Scholar
  50. Picot L, Ravallec R, Fouchereau-Peron M, Vandanjon L, Jaouen P, Chaplain-Derouiniot M, Guérard F, Chabeaud A, Legal Y, Martinez Alvarez O, Berge JP, Piot JM, Batista I, Pires C, Thorkelsson G, Delannoy C, Jakobsen G, Johansson I, Bourseau P (2010) Impact of ultrafiltration and nanofiltration of an industrial fish protein hydrolysate on its bioactive properties. J Sci Food Agric 90:1819–1826Google Scholar
  51. Radha C, Ramesh Kumar P, Prakash V (2008) Preparation and characterization of a protein hydrolysate from an oilseed flour mixture. Food Chem 106:1166–1174CrossRefGoogle Scholar
  52. Schagger H, von Jagow G (1987) Tricine-sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368–379CrossRefGoogle Scholar
  53. See SF, Hong PK, Ng KL, Wan Aida WM, Babji AS (2010) Physicochemical properties of gelatins extracted from skins of different freshwater fish species. Int Food Research J 17:809–816Google Scholar
  54. Segura-Campos MR, Chel-Guerrero LA, Betancur-Ancona DA (2011) Purification of angiotensin I-converting enzyme inhibitory peptides from a cowpea (Vigna unguiculata) enzymatic hydrolysate. Process Biochem 46:864–872CrossRefGoogle Scholar
  55. Thiansilakul Y, Benjakul S, Shahidi F (2007) Compositions, functional properties and antioxidative activity of protein hydrolysates prepared from round scad (Decapterus maruadsi). Food Chem 103:1385–1394CrossRefGoogle Scholar
  56. Tsai JS, Chen JL, Pan BS (2008) ACE-inhibitory peptides identified from the muscle protein hydrolysate of hard clam (Meretrix lusoria). Process Biochem 43:743–747CrossRefGoogle Scholar
  57. Wijesekara I, Qian ZJ, Ryu B, Ngo DH, Kim SK (2011) Purification and identification of antihypertensive peptides from seaweed pipefish (Syngnathus schlegeli) muscle protein hydrolysate. Food Res Int 44:703–707CrossRefGoogle Scholar
  58. Wu J, Aluko RE (2007) Quantitative structure–activity relationship study of bitter di-and tri-peptides including relationship with angiotensin I-converting enzyme inhibitory activity. J Pept Sci 13:63–69CrossRefGoogle Scholar
  59. Wu J, Ding X (2002) Characterization of inhibition and stability of soy protein derived angiotensin I-converting enzyme inhibitory peptides. Food Res Int 35:367–375CrossRefGoogle Scholar
  60. Wu J, Aluko RE, Muir AD (2002) Improved method for direct high performance liquid chromatography assay of angiotensin-converting enzyme catalysed reactions. J Chromatogr A 950:125–130CrossRefGoogle Scholar
  61. 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
  62. Wu H, He HL, Chen XL, Sun CY, Zhang YZ, Zhou BC (2008) Purification and identification of novel angiotensin-I-converting enzyme inhibitory peptides from shark meat hydrolysate. Process Biochem 43:457–461CrossRefGoogle Scholar
  63. Xu W, Kong BH, Zhao XH (2011) Optimization of some conditions of Neutrase-catalyzed plastein reaction to mediate ACE-inhibitory activity in vitro of casein hydrolysate prepared by Neutrase. J Food Sci Technol. doi:10.1007/s13197-011-0503-0
  64. Yang JI, Ho HY, Chu YJ, Chow CJ (2008) Characteristic and antioxidant activity of retorted gelatin hydrolysates from cobia (Rachycentron canadum) skin. Food Chem 110:128–136CrossRefGoogle Scholar
  65. Zhao Y, Li B, Liu Z, Dong S, Zhao X, Zeng M (2007) Antihypertensive effect and purification of an ACE inhibitory peptide from sea cucumber gelatin hydrolysate. Process Biochem 42:1586–1591CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2012

Authors and Affiliations

  • Fatemeh Mahmoodani
    • 1
    • 2
  • Masomeh Ghassem
    • 1
  • Abdul Salam Babji
    • 1
  • Salma Mohamad Yusop
    • 1
  • Roya Khosrokhavar
    • 2
  1. 1.School of Chemical Sciences and Food TechnologyUniversiti Kebangsaan MalaysiaBangiMalaysia
  2. 2.Food and Drug Laboratory Research CenterFood and Drug Organization, MOH & METehranIran

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