Food and Bioprocess Technology

, Volume 12, Issue 5, pp 799–808 | Cite as

Production of Whey-Derived DPP-IV Inhibitory Peptides Using an Enzymatic Membrane Reactor

  • John O’Halloran
  • Michael O’Sullivan
  • Eoin CaseyEmail author
Original Paper


Continuous processing in the production of peptides is an area of increased interest. In this study, an enzymatic membrane reactor (EMR) was developed whereby whey protein isolate was used as a substrate to prepare DPP-IV inhibitory and radical scavenging peptides via enzymatic hydrolysis. Two separate enzymes were tested: Corolase 2TS and Protamex in conventional batch processes and the EMR. Neither enzyme was considered effective at producing peptides with radical scavenging activity when measured using a DPPH assay. However, both enzymes were capable of producing DPP-IV inhibitory peptides. Corolase and Protamex both produced similar DPP-IV inhibition levels upon completion of batch experiments. In the EMR process, permeate in the Protamex run showed 33.7% lower IC50 value compared to the continuous Corolase run. Protamex was a better enzyme at producing the DPP-IV inhibitory effect. The continuous (EMR) production method showed an increased productivity over batch for both enzymes.


Enzymatic membrane reactor DPP-IV inhibitory peptides Peptides with specific bioactivity Whey protein 



Enterprise Ireland is acknowledged for financial support of this research.


  1. Aart, V., Catharina M., Zeeland-Wolbers V., Maria L., Gilst V., Hendrikus W., Nelissen B. and Maria J. (2009). "Egg protein hydrolysates." Patent, WO 128713: 2009.Google Scholar
  2. Adler-Nissen, J. (1986). Enzymic hydrolysis of food proteins. New York: Elsevier Applied Science Publishers.Google Scholar
  3. Brandelli, A., Daroit, D. J., & Corrêa, A. P. F. (2015). Whey as a source of peptides with remarkable biological activities. Food Research International, 73, 149–161.CrossRefGoogle Scholar
  4. Cabrera-Padilla, R. Y., Pinto, G. A., Giordano, R. L., & Giordano, R. C. (2009). A new conception of enzymatic membrane reactor for the production of whey hydrolysates with low contents of phenylalanine. Process Biochemistry, 44(3), 269–276.CrossRefGoogle Scholar
  5. Cheison, S. C., Wang, Z., & Xu, S.-Y. (2006). Hydrolysis of whey protein isolate in a tangential flow filter membrane reactor: I. Characterisation of permeate flux and product recovery by multivariate data analysis. Journal of Membrane Science, 283(1), 45–56.CrossRefGoogle Scholar
  6. Corrêa, A. P. F., Daroit, D. J., Fontoura, R., Meira, S. M. M., Segalin, J., & Brandelli, A. (2014). Hydrolysates of sheep cheese whey as a source of bioactive peptides with antioxidant and angiotensin-converting enzyme inhibitory activities. Peptides, 61, 48–55.CrossRefGoogle Scholar
  7. Eisele, T., Stressler, T., Kranz, B., & Fischer, L. (2013). Bioactive peptides generated in an enzyme membrane reactor using Bacillus lentus alkaline peptidase. European Food Research and Technology, 236(3), 483–490.CrossRefGoogle Scholar
  8. Erdős, B., Grachten, M., Czermak, P., & Kovács, Z. (2018). Artificial neural network-assisted spectrophotometric method for monitoring Fructo-oligosaccharides production. Food and Bioprocess Technology, 11(2), 305–313.CrossRefGoogle Scholar
  9. Ha, G. E., Chang, O. K., Jo, S. M., Han, G. S., Park, B. Y., Ham, J. S., & Jeong, S. G. (2015). Identification of antihypertensive peptides derived from low molecular weight casein hydrolysates generated during fermentation by Bifidobacterium longum KACC 91563. Korean Journal for Food Science of Animal Resources, 35(6), 738–747.CrossRefGoogle Scholar
  10. Haug, A., Hostmark, A. T., & Harstad, O. M. (2007). Bovine milk in human nutrition – A review. Lipids in Health and Disease, 6(1), 25.CrossRefGoogle Scholar
  11. Hernández-Ledesma, B., Recio, I., & Amigo, L. (2008). β-Lactoglobulin as source of bioactive peptides. Amino Acids, 35(2), 257–265.CrossRefGoogle Scholar
  12. ISO F. (2009). Feed products–general guidelines for the determination of nitrogen by the Kjeldahl method. International Organization for Standardiza-tion, Geneva, Switzerland, 1871, 2009.Google Scholar
  13. Jakovetić, S., Luković, N., Jugović, B., Gvozdenović, M., Grbavčić, S., Jovanović, J., & Knežević-Jugović, Z. (2015). Production of antioxidant egg white hydrolysates in a continuous stirred tank enzyme reactor coupled with membrane separation unit. Food and Bioprocess Technology, 8(2), 287–300.CrossRefGoogle Scholar
  14. Janson, J.-C. (2012). Protein purification: Principles, high resolution methods, and applications, John Wiley & Sons.Google Scholar
  15. Jemil, I., Jridi, M., Nasri, R., Ktari, N., Salem, R. B. S.-B., Mehiri, M., Hajji, M., & Nasri, M. (2014). Functional, antioxidant and antibacterial properties of protein hydrolysates prepared from fish meat fermented by Bacillus subtilis A26. Process Biochemistry, 49(6), 963–972.CrossRefGoogle Scholar
  16. Juillerat-Jeanneret, L. (2013). Dipeptidyl peptidase IV and its inhibitors: Therapeutics for type 2 diabetes and what else? Journal of Medicinal Chemistry, 57(6), 2197–2212.CrossRefGoogle Scholar
  17. Kamau, S. M., & Lu, R.-R. (2011). The effect of enzymes and hydrolysis conditions on degree of hydrolysis and DPPH radical scavenging activity of whey protein hydrolysates. Current Research in Dairy Sciences, 3, 25–35.CrossRefGoogle Scholar
  18. Lacroix, I. M., Meng, G., Cheung, I. W., & Li-Chan, E. C. (2016). Do whey protein-derived peptides have dual dipeptidyl-peptidase IV and angiotensin I-converting enzyme inhibitory activities? Journal of Functional Foods, 21, 87–96.CrossRefGoogle Scholar
  19. Le Maux, S., Nongonierma, A. B., Barre, C., & FitzGerald, R. J. (2016). Enzymatic generation of whey protein hydrolysates under pH-controlled and non pH-controlled conditions: Impact on physicochemical and bioactive properties. Food Chemistry, 199, 246–251.CrossRefGoogle Scholar
  20. Le Maux, S., Nongonierma, A. B., & FitzGerald, R. J. (2015a). Improved short peptide identification using HILIC–MS/MS: Retention time prediction model based on the impact of amino acid position in the peptide sequence. Food Chemistry, 173, 847–854.CrossRefGoogle Scholar
  21. Le Maux, S., Nongonierma, A. B., Murray, B., Kelly, P. M., & FitzGerald, R. J. (2015b). Identification of short peptide sequences in the nanofiltration permeate of a bioactive whey protein hydrolysate. Food Research International, 77, 534–539.CrossRefGoogle Scholar
  22. Manders, R. J., Hansen, D., Zorenc, A. H., Dendale, P., Kloek, J., Saris, W. H., & van Loon, L. J. (2014). Protein co-ingestion strongly increases postprandial insulin secretion in type 2 diabetes patients. Journal of Medicinal Food, 17(7), 758–763.CrossRefGoogle Scholar
  23. Mateo, C., Palomo, J. M., Fernandez-Lorente, G., Guisan, J. M., & Fernandez-Lafuente, R. (2007). Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme and Microbial Technology, 40(6), 1451–1463.CrossRefGoogle Scholar
  24. Méric, E., Lemieux, S., Turgeon, S. L., & Bazinet, L. (2014). Insulin and glucose responses after ingestion of different loads and forms of vegetable or animal proteins in protein enriched fruit beverages. Journal of Functional Foods, 10, 95–103.CrossRefGoogle Scholar
  25. Nath, A., Verasztó, B., Basak, S., Koris, A., Kovács, Z., & Vatai, G. (2016). Synthesis of lactose-derived nutraceuticals from dairy waste whey—A review. Food and Bioprocess Technology, 9(1), 16–48.CrossRefGoogle Scholar
  26. Noble, R. D. and S. A. Stern (1995). Membrane separations technology: Principles and applications, Elsevier.Google Scholar
  27. Nongonierma, A. B., & FitzGerald, R. J. (2013a). Dipeptidyl peptidase IV inhibitory and antioxidative properties of milk protein-derived dipeptides and hydrolysates. Peptides, 39, 157–163.CrossRefGoogle Scholar
  28. Nongonierma, A. B., & FitzGerald, R. J. (2013b). Inhibition of dipeptidyl peptidase IV (DPP-IV) by proline containing casein-derived peptides. Journal of Functional Foods, 5(4), 1909–1917.CrossRefGoogle Scholar
  29. Nongonierma, A. B., & FitzGerald, R. J. (2013c). Inhibition of dipeptidyl peptidase IV (DPP-IV) by tryptophan containing dipeptides. Food & Function, 4(12), 1843–1849.CrossRefGoogle Scholar
  30. Nongonierma, A. B., & FitzGerald, R. J. (2016). Prospects for the management of type 2 diabetes using food protein-derived peptides with dipeptidyl peptidase IV (DPP-IV) inhibitory activity. Current Opinion in Food Science, 8, 19–24.CrossRefGoogle Scholar
  31. Peng, X., Kong, B., Xia, X., & Liu, Q. (2010). Reducing and radical-scavenging activities of whey protein hydrolysates prepared with Alcalase. International Dairy Journal, 20(5), 360–365.CrossRefGoogle Scholar
  32. Peng, X., Xiong, Y. L., & Kong, B. (2009). Antioxidant activity of peptide fractions from whey protein hydrolysates as measured by electron spin resonance. Food Chemistry, 113(1), 196–201.CrossRefGoogle Scholar
  33. Rodrigues, R. C., Ortiz, C., Berenguer-Murcia, Á., Torres, R., & Fernández-Lafuente, R. (2013). Modifying enzyme activity and selectivity by immobilization. Chemical Society Reviews, 42(15), 6290–6307.CrossRefGoogle Scholar
  34. Seader, J. D. and E. J. Henley (2011). "Separation process principles." John Wiley and Sons, Inc.Google Scholar
  35. Solieri, L., Rutella, G. S., & Tagliazucchi, D. (2015). Impact of non-starter lactobacilli on release of peptides with angiotensin-converting enzyme inhibitory and antioxidant activities during bovine milk fermentation. Food Microbiology, 51, 108–116.CrossRefGoogle Scholar
  36. Spellman, D., McEvoy, E., O’cuinn, G., & FitzGerald, R. (2003). Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. International Dairy Journal, 13(6), 447–453.CrossRefGoogle Scholar
  37. Tong, L. M., Sasaki, S., McClements, D. J., & Decker, E. A. (2000). Mechanisms of the antioxidant activity of a high molecular weight fraction of whey. Journal of Agricultural and Food Chemistry, 48(5), 1473–1478.CrossRefGoogle Scholar
  38. Van Reis, R., & Zydney, A. (2001). Membrane separations in biotechnology. Current Opinion in Biotechnology, 12(2), 208–211.CrossRefGoogle Scholar
  39. Zambrowicz, A., Polanowski, T. A., Lubec, G., & Trziszka, T. (2013). Manufacturing of peptides exhibiting biological activity. Amino Acids, 44(2), 315–320.CrossRefGoogle Scholar
  40. Zhu, C.-F., Li, G.-Z., Peng, H.-B., Zhang, F., Chen, Y., & Li, Y. (2010). Treatment with marine collagen peptides modulates glucose and lipid metabolism in Chinese patients with type 2 diabetes mellitus. Applied Physiology Nutrition and Metabolism, 35(6), 797–804.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Chemical and Bioprocess EngineeringUniversity College Dublin (UCD)Dublin 4Ireland
  2. 2.UCD Institute of Food & Health, School of Agriculture and Food ScienceUniversity College DublinDublin 4Ireland

Personalised recommendations