Antioxidant Activity of Hydrolysates Prepared from Flaxseed Cake Proteins Using Pancreatin


Proteins were isolated from defatted flaxseed cake and hydrolysed with pancreatin. The hydrolysis process was conducted at a stable temperature of 50°C and pH 7.5, and monitored with the pH-stat method. The obtained hydrolysates with a degree of hydrolysis (DH) of 5, 10, 15, 20, 25% were investigated in terms of antioxidant properties. The radical scavenging activity was assayed against DPPH· and ABTS·+, the reducing ability - with FRAP assay, and the capability to bind Fe(II) - by reaction with ferrozine. SE-HPLC analysis was used to determine molecular weight distribution of hydrolysis products.

The antiradical activity of pancreatin hydrolysates of flaxseed proteins was increasing along with an increasing DH and for the hydrolysate with DH 25% the EC50 value determined with the DPPH assay accounted for 0.083 mg/assay, and the ABTS·+ scavenging activity - for 0.218 mmol Trolox/g. This hydrolysate was constituted mainly by peptides with low molecular weights (MW) of 0.238-0.556 kDa. In turn, the Fe(II) binding capability increased from 44.5% to 64.9% in the case of hydrolysates with DH 5-20% and decreased in the case of the hydrolysate with DH 25%. A similar de-pendency was observed in the ability of pancreatin hydrolysates of flaxseed proteins to reduce Fe(III). The maximum value of reducing ability reached 0.25 mmol Fe(II)/g for the hydrolysate with DH 20% that was predominated by polypeptides and peptides with MW of 0.238-1.046 Da.

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  • 1. Adler-Nissen J., Enzymatic Hydrolysis of Food Proteins. 1986, 1st ed., Elsevier Applied Science, London, pp. 57-68; 132-188.

  • 2. Ajibola C.F., Fashakin J.B., Fagbemi T.N., Aluko R.E., Effect of peptide size on antioxidant properties of African yam bean seed (Sphenostylis stenocarpa) protein hydrolysate fractions. Int. J. Mol. Sci., 2011, 12, 6685-6702.

  • 3. Alashi A.M., Blanchard CL., Mailer R.J., Agboola S.O., Maw-son AJ., He R, Girgih A., Aluko RE., Antioxidant properties of Australian canola meal protein hydrolysates. Food Chem., 2014, 146, 500-506.

  • 4. Andriamihaja M., Guillot A., Svendsen A., Hagedorn J., Rako-tondratohanina S., Tome D., Blachier F., Comparative effciency of microbial enzyme preparations versus pancreatin for in vitro alimentary protein digestion. Amino Acids, 2013, 44, 563-572.

  • 5. AOAC, Offcial Methods of Analysis, 1990, 15th ed., Arling-ton Virginia, USA.

  • 6. Benzie I.F.F., Strain J.J., The ferric reducing ability of plasma (FRAP) as a measure of ‘’antioxidant power’’: The FRAP assay Anal. Biochem., 1996, 239, 70-76.

  • 7. Betancur-Ancona D., Sosa-Espinoza T., Ruiz-Ruiz J., Segura–Campos M., Chel-Guerrero L., Enzymatic hydrolysis of hard–to-cook bean (Phaseolus vulgaris L.) protein concentrates and its effects on biological and functional properties. Int. J. Food Sci. Tech., 2014, 49, 2-8.

  • 8. Borg D.C., Oxygen free radicals and tissue injury. 1993, in: Oxy-gen Free Radicals in Tissue Damage (eds. M. Tarr, F. Samson). Birkhäuser, Boston, MA, USA, pp. 12-53.

  • 9. Brand-Williams W, Cuvelier M.E., Berset C, Use of a free–radical method to evaluate antioxidant activity. Lebensm. Wiss. Technol., 1995, 28, 25-30.

  • 10. Dev D.K., Quensel E., Preparation and functional-properties of linseed protein products containing differing levels of muci-lage. J. Food Sci., 1988, 53, 1834-1837.

  • 11. Hartmann R, Meisel H., Food-derived peptides with biological activity: from research to food applications. Curr. Opin. Biotech., 2007, 18, 163-169.

  • 12. Karamać M., Pegg RB., Limitations of the tetramethylmurexide assay for investigating the Fe(II) chelation activity of phenolic compounds. J. Agric. Food Chem., 2009, 57, 6425-6431.

  • 13. Karamać M., Amarowicz R, Kostyra H., Effect of temperature and enzyme/substrate ratio on the hydrolysis of pea protein iso-lates by trypsin. Czech J. Food Sci., 2002, 20, 1-6.

  • 14. Karamać M., Flaczyk E., Wanasundara P.K.J.P.D., Amarowicz R., Angiotensin I-converting enzyme (ACE) inhibitory activity of hydrolysates obtained from muscle food industry by-prod-ucts - a short report. Pol. J. Food Nutr. Sci., 2005, 55, 133-138.

  • 15. Korhonen H., Pihlanto A., Bioactive peptides: Production and functionality. Int. Dairy J., 2006, 16, 945-960.

  • 16. Li Y.H., Jiang B., Zhang T., Mu W.M., Liu J., Antioxidant and free radical-scavenging activities of chickpea protein hydro-lysate (CPH). Food Chem., 2008, 106, 44-450.

  • 17. Ma Y.Y., Xiong, Y.L.L., Zhai J.J., Zhu H.N., Dziubla T., Fractionation and evaluation of radical scavenging peptides from in vitro digests of buckwheat protein. Food Chem., 2010, 118, 582-588.

  • 18. Marambe PWM.L.H.K., Shand P.J., Wanasundara J.P.D., An in-vitro investigation of selected biological activities of hydro-lysed faxseed (Linum usitatissimum L.) proteins. J. Am. Oil Chem. Soc, 2008, 85, 1155-1164.

  • 19. Marambe H.K., Shand P.J., Wanasundara J.P.D., Release of an-giotensin I-converting enzyme inhibitory peptides from faxseed (Linum usitatissimum L.) protein under simulated gastrointesti-nal digestion. J. Agric. Food Chem., 2011, 59, 9596-9604.

  • 20. Moller N.P, Scholz-Ahrens K.E., Roos N, Schrezenmeir J., Bio-active peptides and proteins from foods: indication for health ef-fects. Eur. J. Nutr., 2008, 47, 171-182.

  • 21. Mueller K., Eisner P, Yoshie-Stark Y, Nakada R., Kirchhoff E., Functional properties and chemical composition of fractionated brown and yellow linseed meal (Linum usitatissimum L.). J. Food Eng., 2010, 98, 453-460.

  • 22. Ng K.L., Ayob M.K., Said M., Osman M.A., Ismail A., Optimization of enzymatic hydrolysis of palm kernel cake protein (PKCP) for producing hydrolysates with antiradical capacity. Ind. Crop Prod., 2013, 43, 725-731.

  • 23. Oomah B.D., Mazza G., Flaxseed proteins - a review. Food Chem., 1993, 48, 109-114.

  • 24. Panasiuk R, Amarowicz R, Kostyra H., Sijtsma L., Determination of a-amino nitrogen in pea protein hydrolysates: a compari-son of three analytical methods. Food Chem., 1998, 62, 363-367.

  • 25. Popovic L., Pericin D., Vastag Z., Popovic S., Krimer V, Torbica A., Antioxidative and functional properties of pumpkin oil cake globulin hydrolysates. J. Am. Oil Chem. Soc, 2013, 90, 1157-1165.

  • 26. Pownall T.L., Udenigwe CC, Aluko RE., Amino acid composition and antioxidant properties of pea seed (Pisum sativum L.) enzymatic protein hydrolysate fractions. J. Agric. Food Chem., 2010, 58, 4712-4718.

  • 27. Rabetafka H.N., Van Remoortel V, Danthine S., Paquot M., Blecker C, Flaxseed proteins: food uses and health benefts. Int. J. Food Sci. Tech., 2011, 46, 221-228.

  • 28. Re R, Pellegrini N, Proteggente A., Pannala A., Yang M., Rice–Evans C, Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med., 1999, 26, 1231-1237.

  • 29. Samaranayaka A.G.P, Li-Chan, E.C.Y, Food-derived peptidic antioxidants: A review of their production, assessment, and po-tential applications. J. Funct. Food, 2011, 3, 229-254.

  • 30. Silva F.G.D., O'Callagahan Y., O'Brien N.M., Netto FM., Anti-oxidant capacity of faxseed products: the effect of in vitro digestion. Plant Food Hum. Nutr., 2013, 68, 24-30.

  • 31. Udenigwe CC, Aluko R.E., Antioxidant and angiotensin con-verting enzyme-inhibitory properties of a faxseed protein-de-rived high fscher ratio peptide mixture. J. Agric. Food Chem., 2010, 58, 4762-4768.

  • 32. Udenigwe CC, Lin Y.-S., Hou W.-C, Aluko R.E., Kinetics of the inhibition of renin and angiotensin I-converting enzyme by faxseed protein hydrolysate fractions. J. Funct. Foods, 2009a, 1, 199-207.

  • 33. Udenigwe CC, Lu Y.L., Han C.H., Hou W.C, Aluko R.E., Flaxseed protein-derived peptide fractions: Antioxidant properties and inhibition of lipopolysaccharide-induced nitric oxide production in murine macrophages. Food Chem., 2009b, 116, 277-284.

  • 34. Valdez-Ortiz A., Fuentes-Gutierrez CI., German-Baez L.J. Gutierrez-Dorado R, Medina-Godoy S., Protein hydrolysates obtained from Azufrado (sulphur yellow) beans (Phaseo-lus vulgaris): Nutritional, ACE-inhibitory and antioxidative characterization. LWT-Food Sci. Technol., 2012, 46, 91-96.

  • 35. Zambrowicz A., Timmer M., Eckert E., Trziszka T, Evaluation of the ACE-inhibitory activity of egg-white proteins degraded with pepsin. Pol. J. Food Nutr. Sci., 2013, 63, 103-108.

  • 36. Zhang M.-N, Huang G.-R, Jiang J.-X., Iron binding capacity of dephytinised soy protein isolate hydrolysate as infuenced by the degree of hydrolysis and enzyme type. J. Food Sci. Tech., 2014, 51, 994-999.

  • 37. Zheng L., Ren J., Su G, Yang B., Zhao M., Comparison of in vitro digestion characteristics and antioxidant activity of hot- and cold–pressed peanut meals. Food Chem., 2013, 141, 4246-4252.

  • 38. Zilic S., Akillioglu G., Serpen A., Barac M., Gokmen V, Ef-fects of isolation, enzymatic hydrolysis, heating, hydratation and Maillard reaction on the antioxidant capacity of cereal and legume proteins. Food Res. Int., 2012, 49, 1-6.


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