VEGETOS: An International Journal of Plant Research & Biotechnology
(Society For Plant Research)

Research Articles


Volume: 32, Issue: 3, September 2019

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Page Visits: 33

Doi: 10.1007/s42535-019-00031-6
Doi Link:
First Page: 363
Last Page: 369
Published: 20 June, 2019

Enzymatic treatment improves ACE-I inhibiton and antiproliferative potential of chickpea


Chickpea seeds are the preferred source of proteins possessing health care functions in countries across the world. Study indicated the chickpea proteins as a promising center of bioactive peptides and open up new vista for food industry. Employing gastrointestinal enzyme alcalase, protein hydrolysates generated from 45 chickpea seed accessions were evaluated for angiotensin-I converting enzyme (ACE-I) inhibitory potential and antiproliferative influence. Alcalase at  1 h of optimum hydrolysis produced bioactive peptides inhibiting the ACE-I activity. The accession BDN-9-3 gave highest ACE-I inhibitory activity with IC50 value of 22.43 mg/ml. The protein hydrolysate of BDN-9-3 was further subjected to antiproliferative assessment against breast cancer cells MCF-7 and MDA-MB-231. The IC50 of BDN-9-3 alcalase hydrolysate was 0.60 mg/ml and 0.63 mg/ml in MCF-7 and MDA-MB-231 cells respectively, compared to non hydrolyzed chickpea protein (IC50 of 0.85 and 0.82 mg/ml). Present study ascertain that chickpea seed hydrolysate can be perceived as a valuable nutraceutical resource.



ACE-I inhibitory activity, Chickpea seed proteins, Alcalase


  1. Boschin G, Scigliuolo GM, Resta D, Arnoldi A (2014) ACE-inhibitory activity of enzymatic protein hydrolysates from lupin and other legumes. Food Chem 145:34–40.

  2. Boyer PD (ed) (1973) The enzyme. Academic Press, New York, p 650

  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254.

  4. Cancer (2014) I. A. f. R. o. World cancer report. WHO, Geneva

  5. Chitra U, Singh U, Rao PV (1996) Phytic acid, in vitro protein digestibility, dietary fiber, and minerals of pulses as influenced by processing methods. Plant Foods Hum Nutr 49:307–316

  6. Cushman DW, Cheung HS (1971) Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 20:1637–1648.

  7. Daskaya DC, Yucetepe A, Karbancioglu GF, Daskaya H, Ozcelik B (2017) Angiotensin-I-converting enzyme (ACE) inhibitory peptides from plants. Nutrients 23:9(4).

  8. De Castro RJS, Sato HH (2014) Antioxidant activities and functional properties of soy protein isolate hydrolysates obtained using microbial proteases. Int J Food Sci Tech 49:317–328.

  9. Doke S, Guha M (2015) Nutritional, physico-chemical and functional properties of ready-to-use chickpea and soybean flour. Int J Food Nutr Sci 4:2320–7876

  10. Erdmann K, Cheung BWY, Schroder H (2008) The possible roles of food-derived bioactive peptides in reducing the risk of cardiovascular disease. J Nutr Biochem 19:643–654.

  11. Gautam AK, Gupta N, Narvekar DT, Bhadkariya R, Bhagyawant SS (2018) Characterization of chickpea (Cicer arietinum L.) lectin for biological activity. Physiol Mol Biol Plants 24(3):389–397.

  12. Gupta N, Shrivastava N, Bhagyawant SS (2017) Multivariate analysis based on nutritional value, antinutritional profile and antioxidant capacity of forty chickpea genotypes grown in India. J Nutr Food Sci.

  13. Gupta N, Bisen PS, Bhagyawant SS (2018) Chickpea lectin inhibits human breast cancer cell proliferation and induces apoptosis through cell cycle arrest. Protein Pept Lett 25:492–499.

  14. Hong LG, Wei LG, Liu H, Hui SY (2005) Mung-bean protein hydrolysates obtained with alcalase exhibit angiotensin I-converting enzyme inhibitory activity. Food Sci Technol Inter 11:281–287.

  15. Hsu HW, Vavak DL, Satterlee LD, Miller GA (1977) A multienzyme technique for estimating protein digestibility. J Food Sci 42:1269–1273.

  16. Hua ZY, Jun GJ, Hua LZ, Jing T (2011) Research progress of ACE inhibitory peptide. Cereals Oils 25:44–46

  17. Hyun CK, Shin HK (2000) Utilization of bovine blood plasma proteins for the production of angiotensin I-converting enzyme inhibitory peptides. Process Biochem 36:65–71.

  18. Jakubczyk A, Baraniak B (2013) Activities and sequences of the angiotensin I- converting enzyme (ACE) inhibitory peptides obtained from the digested lentil (Lens culinaris) globulins. Inter J Food Sci Tech 44:2363–2369.

  19. 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 49:2992–2997.

  20. Kou X, Gao J, Xue Z, Zhang Z, Wang H, Wang X (2013) Purification and identification of antioxidant peptides from chickpea (Cicer arietinum L.) albumin hydrolysates. LWT Food Sci Tech 50:591–598.

  21. 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 Tech 222:733–736.

  22. Markland FS, Smith EL (1971) Subtilisins: primary structure, chemical and physical properties. Enzymes 3:561–608.

  23. Matsui T, Matsufuj H, Seki E, Osajima K, Nakashima M, Osajima Y (1993) Inhibition of angiotens in I-converting enzyme by Bacillus licheniformis alkaline protease hydrolyzate derived from sardine muscle. Biosci Biotech Biochem 57:922–925.

  24. Mohammad Z, Bita F, Afshin E, Azizah AH, Farooq A, Nazamid S (2015) In vitro and in vivo antihypertensive activity of palm kernel cake protein hydrolysates: sequencing and characterization of potent bioactive peptides. Ind Crops Prod 76:112–120.

  25. Pedroche J, Yust MM, Girón-Calle J, Alaiz M, Millán F, Vioque J (2002) Utilisation of chickpea protein isolates for production of peptides with angiotensin I-converting enzyme (ACE)-inhibitory activity. J Sci Food Agric 82:960–965.

  26. Singh U, Jambunathan R (1981) Studies on desi and kabuli chickpea (Cicer arietinum L.) cultivars: levels of protease inhibitors, levels of polyphenolic compounds and in vitro protein digestibility. J Food Sci 46:1364–1367.

  27. Skehan P et al (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Institute 82:1107–1112.

  28. Vermeirssen V, Van Camp J, Devos L, Verstraete W (2003) Release of angiotensin I converting enzyme (ACE) inhibitory activity during in vitro gastrointestinal digestion: from batch experiment to semi continuous model. J Agric Food Chem 51:5680–5687.

  29. Wang FJ, Yin XY, Regenstein JM, Wang JZ (2016) Separation and purification of angiotensin-I-converting enzyme (ACE) inhibitory peptides from walnuts (Juglans regia L.) meal. Eur Food Res Tech 242:911–918.

  30. Wu WL, Wu GJ, Liang DS, Yang F (2006) The physiological function and research progress of angiotensin-I-converting enzyme inhibitory peptides. Mod Food Sci Tech 22:251–254

  31. Yust MM, Pedroche J, Girón-Calle J, Alaiz M, Millán F, Vioque J (2003) Production of ace inhibitory peptides by digestion of chickpea legumin with alcalase. Food Chem 81:363–369.

  32. Zhang T, Li YH, Miao M, Jiang B (2011) Purification and characterization of a new antioxidant peptide from chickpea (Cicer arietinum L.) protein hydrolysates. Food Chem 128:28–33.

  33. Zheng Y, Li Y, Zhang Y, Ruan X, Zhang R (2017) Purification, characterization, synthesis, in vitro ACE inhibition and in vivo antihypertensive activity of bioactive peptides derived from oil palm kernel glutelin-2 hydrolysate. J Funct Foods 28:48–58.



Acknowledgements :

The authors are thankful to the Hon’ble Vice-chancellor Prof. Sangeeta Shukla, Jiwaji University, Gwalior, India for providing research grant.

Author Information:

Neha Gupta
School of Studies in Biotechnology, Jiwaji University, Gwalior, India

Sameer Suresh Bhagyawant
School of Studies in Biotechnology, Jiwaji University, Gwalior, India

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