Kidney therapeutic potential of peptides derived from the bromelain hydrolysis of green peas protein

Document Type: Original Article


1 Faculty of Medicine, Universitas Kristen Maranatha, Bandung Indonesia

2 School of Pharmacy, Institute Teknologi Bandung Indonesia

3 Alphasains Centre, Serpong Indonesia

4 Faculty of Medicine, Universitas Jenderal Achmad Yani, Bandung Indonesia


Objective(s): Kidney disease is a global health problem that needs a solution to its therapy. In the previous study, we found that protein hydrolysate of green peas origin of Indonesia hydrolysed by bromelain (PHGPB) showed improve kidney function in cisplatin-induced nephropathy rats. In this study, we investigated the effect of PHGPB to obtain effective dose that exerts a therapeutic effect on chronic kidney disease (CKD) based on reducing urea and creatinine levels and to elucidate its mechanism of action.
Materials and Methods: Two sets of experiments were conducted: (1) characteristics and  proteomic profile of PHGPB, (2) in vivo test of PHGPB in gentamycin-induced Wistar rats, including urea and creatinine measurements, activities of antioxidant and kidney-related peptides (ANP, COX-1, and renin).
Results: PHGPB showed three bands under 10 kDa using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and contained 10 identified proteins using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Significant differences in urea and creatinine levels were found between all PHGPB treatments and positive controls (P<0.01). The lowest levels of urea and creatinine that were validated by high super oxide dismutase (SOD) activity and atrial natriuretic peptide (ANP) level were obtained in the 200 mg/day PHGPB treatment. However, the mean renin level was high and cyclooxygenase-1 (COX-1) level did not exceed positive and negative control levels.
Conclusion: PHGPB at dose 200 mg/kgBW shows a potential CKD therapeutic effect that is dose-dependent. Higher PHGPB dose corresponds to better effect on kidney function by increasing antioxidant activity and ANP levels in gentamycin-induced Wistar rats.


Main Subjects

1. Badan penelitian dan pengembangan kesehatan indonesia. Riset Kesehatan Dasar (RISKESDAS). Health Res Dev Agency Indonesia 2013; 2013:1-384.
2. Hoskins I. Pea protein may prevent kidney disease [Internet]. 2017. Available from:
3. Korhonen H, Pihlanto  A. Food-derived bioactive peptides-opportunities for designing future foods. Curr Pharm Des 2003; 9:1297-1308.
4. Malomo SA, Aluko RE. A comparative study of the structural and functional properties of isolated hemp seed (Cannabis sativa L.) albumin and globulin fractions. Food Hydrocoll 2015; 43:743-752.
5.  Tavano OL. Protein hydrolysis using proteases: An important tool for food biotechnology. J Mol Catal B Enzym 2013; 90:1-11.
6.  Pownall TL, 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.
7. Shahidi F, Zhong Y. Bioactive peptides. J AOAC Int 2008; 91:914-931.
8. Kim SK, Wijesekara I. Development and biological activities of marine-derived bioactive peptides: A review. J Funct Foods 2010; 2:1-9.
9. Li H, Aluko RE. Identification and inhibitory properties of multifunctional peptides from pea protein hydrolysate. J Agric Food Chem 2010; 58:11471–11476.
10. Pownall TL, Udenigwe CC, Aluko RE. Effects of cationic property on the in vitro antioxidant activities of pea protein hydrolysate fractions. Food Res Int 2011; 44:1069-1074.
11. Gajanan PG, Elavarasan K, Shamasundar BA. Bioactive and functional properties of protein hydrolysates from fish frame processing waste using plant proteases. Environ Sci Pollut Res 2016; 23:24901-24911.
12. Sherwood L. Human Physiology: From Cells to Systems. 7th ed. Human Physiology. Yolanda Cossio; 2010. p 766.
13. Santos-Araújo, C., Leite-Moreirab,A., Pestana M. Clinical value of natriuretic peptides in chronic kidney disease. Nefrologia 2015; 35:227-233.
14. Aluko RE. Structure and function of plant protein-derived antihypertensive peptides. Curr Opin Food Sci 2015; 4:44-50.
15. Levin ER, Gardner DG, Samson WK. Natriuretic peptides. N Engl J Med 1998;  339:321-328.
16. Saito Y. Roles of atrial natriuretic peptide and its therapeutic use. J Cardiol 2010; 56:262-270.
17. Ogawa N, Komura H, Kuwasako K, Kitamura K, Kato J. Plasma levels of natriuretic peptides and development of chronic kidney disease. BMC Nephrol 2015; 16:171-177.
18. de Chatel R, Mako J, Toth M, Barna I, Lang RE. Atrial natriuretic peptide (ANP) in patients with chronic renal failure on maintenance haemodialysis. Int Urol Nephrol  1991; 23:177-183.
19. Hidayat M. Preparation and Examination of Hydrolysate Protein of Green Peas by bromelain for Improvement Kidney Function. Indonesia; EC00201810615, 2018.
20. Kunitz M. Crystalline Deoxyribonuclease I. Isolation and general properties spectrophotometric method fo the measurement of desoxyribonuclease activity. J Genet Physiol 1950; 33:349-362.
21. Harlow E, Lane D. Bradford Assay. Cold Spring Harb Protoc [Internet]. 2006;2006(6):pdb.prot4644-pdb.prot4644. Available from:
22. Alu’datt MH, Rababah T, Alhamad MN, Alodat M, Al-Mahasneh MA GS. Molecular characterization and bio-functional property determination using SDS-PAGE and RP-HPLC of protein fractions from two Nigella species. Food Chem 2017; 230:125-134.
23. Laemmli UK. Cleavage of structural proteins during the assembly of bacteriophage T4. Nature 1970; 227:680-685.
24. Li H, Prairie N, Udenigwe CC, Adebiyi AP, Tappia PS, Aukema HM, et al. Blood pressure lowering effect of a pea protein hydrolysate in hypertensive rats and humans. J Agric Food Chem 2011; 59:9854-9860.
25. Restriani R. Hidrolisis secara enzimatis protein bungkil biji ryamplung (Calophyllum inophyllum) menggunakan bromelain. Biota 2015; 1:86-91.
26. Hale LP, Greer PK, Trinh CT. Proteinase activity and stability of natural bromelain preparations. Int Immunopharmacol 2005; 5:783-793.
27. Poh SS and Abdul Majid F. Thermal stability of free bromelain and bromelain-plyphenol complex in pineapple juice. Int Food Res J 2011; 18:1051-1060.
28. Carrillo E, Rubiales D, Castillejo MA. Proteomic Analysis of Pea (Pisum sativum L.) Response During Compatible and Incompatible Interactions with the Pea Aphid (Acyrthosiphon pisum H.). Plant Mol Biol Rep 2014; 32:697-718.
29. Giknis MLA CC. Clinical Laboratory Parameters for Crl: WI (Han). Montreal: Charles River Accelerating drug development; 2008. p 9.
30. Pavan R, Jain S, Shraddha, Kumar A. Properties and therapeutic application of bromelain: a review. Biotechnol Res Int 2012; 2012:976203-976209.  
31. Girgih AT, Nwachukwu ID, Onuh JO, Malomo SA, Aluko RE. Antihypertensive properties of a pea protein hydrolysate during short- and long-term oral administration to spontaneously hypertensive rats. J Food Sci 2016; 81:1281-1287.
32. Sanchmonge R, Lopez-Torrejon G, Pascual CY, Varela J, Martin-Esteban M SG. Vicilin and convicilin are potential major allergens from pea. Clin Exp Allergy 2004; 34:1747-1783.
33. Gilman DC, Peck HT CF. Legumin. In: Rines GE, editor. The Encyclopedia Americana . New York: Dodd, Mead; 1920.
34. Leterme P. Recommendations by health organizations for legume consumption. Br J Nutr 2002; 88:239-242.
35. Rubio LA, Pérez A, Ruiz R, Guzmán MÁ, Aranda-Olmedo I, Clemente A. Characterization of pea (Pisum sativum) seed protein fractions. J Sci Food Agric 2014; 94:280-287.
36. Maphosa, Y., Jideani V. Chapter 6: The role of legumes in
human nutrition. In: Functional food – improve health through adequate food. I 2017. Intechopen Publisher, pp 103-121.
37. Lopez-Novoa JM, Quiros Y, Vicente L, Morales AI, Lopez-Hernandez FJ. New insights into the mechanism of aminoglycoside nephrotoxicity: An integrative point of view. Kidney Int 2011; 79:33-45.
38. Stanisavljević NS, Goran N. Vukotić GN , Pasto FT3, Sužnjević D JZ. antioxidant activity of pea protein hydrolysates produced by batch fermentation with lactic acid bacteria. Arch Biol Sci 2015; 67:1033-1042.
39. Badr G, Ramadan NK, Sayed LH, Badr BM, Omar HM, Selamoglu Z. Camel whey protein as a new dietary approach to the management of free radicals and for the treatment of different health disorders. Iran J Basic Med Sci 2017; 20:338-349.
40. Douglas C. Eaton JPP. Vanders Renal Physiology. 8th ed. Lange -Mc Graw Hill; Cited: October 30, 2018.
41. Kasama S, Furuya M, Toyama T, Ichikawa S, Kurabayashi M. Effect of atrial natriuretic peptide on left ventricular remodelling in patients with acute myocardial infarction. Eur Hear J 2008; 29:1485-1494.
42. Gutkowska J, Jankowski M, Antunes-Rodrigues J. The role of oxytocin in cardiovascular regulation. Am J Physiol Regul Integr Comp Physiol 2007; 293:267-275.