• Lida Ebrahimi
  • K. A. Ayub Nawaz
  • K. G. Kiran
  • S. Durairaju Nisshanthini
  • Muthusamy Palaniswamy
  • Jayaraman Angayarkanni Department of Microbial Biotechnology, Bharathiar University, Coimbatore (T. N) 641046, India


Objective: The present study was planned to explore safer, innovative and economic Angiotensin-converting enzyme inhibitors (ACEi) from beef extract by the action of a proteolytic Micrococcus luteus. Cytotoxicity of the stable peptide was predicted using MCF-7 cell line in vitro.

Methods: ACEi was purified by sequential steps of ethanol precipitation, ion exchange column chromatography (MonoQ) and gel filtration column chromatography (Sephadex G25). The apparent molecular mass was determined by SDS-PAGE. The anticancer property was analyzed by studying the cytotoxicity effects of angiotensin converting enzyme inhibitor using Breast cancer MCF-7 cell lines

Results: The peptide was purified and molecular mass was determined as 4.5 kDa. The IC50 value of peptide was found to be 59.5 µg/ml. The DNA fragmentation was not observed in the treated cells. The purified peptide has demonstrated to induce apoptosis of cancer cell. The results proved that the peptide has the ability to be used for cancer therapy.

Conclusion: The presence of ACE inhibition activities in the fermentation of beef extract using Micrococcus luteus has been investigated. The Peptide has been determined as an active compound that inhibited the activity of ACE. These properties indicate the possibilities of the use of purified protein as a potent anticancer agent.

Keywords: Angiotensin-converting enzyme inhibitors, Micrococcus luteus, Anti-proliferative, Anti-metastatic, MCF-7 cell line, Anticancer activity.


Download data is not yet available.


Khurana R, Simons M, Martin JF, Zachary IC. Role of angiogenesis in cardiovascular disease: a critical appraisal. Circulation 2005;112:1813–24.

Noguchi R, Yoshiji H, Kuriyama S, Yoshii J, Ikenaka Y, Yanase K, et al. Combination of interferon-beta and the angiotensin-converting enzyme inhibitor, perindopril, attenuates murine hepatocellular carcinoma development and angiogenesis. Clin Cancer Res 2003;9:6038–45.

Yasumatsu R, Torahiko N, Muneyuki M, Aya I, Yuichiro K, Takashi N, et al. Effects of the angiotensin-I converting enzyme inhibitor perindopril on tumor growth and angiogenesis in head and neck squamous cell carcinoma cells. J Cancer Res Clin Oncol 2004;130:567–73.

Volpert O, Ward WF, Lingen MW, Chesler L, Solt DB, Johnson MD, et al. Captopril inhibits angiogenesis and slows the growth of experimental tumor in rats. J Clin Invest 1996;98:671-9.

Ryan JT, Ross RP, Bolton D, Fitzgerald GF, Stanton C. Bioactive peptides from muscle sources: meat and fish. Nutrients 2011;3:765-91.

Korhonen H, Pihlanto-Leppala A. Milk protein-derived bioactive peptides-novel opportunities for health promotion. IDF Bull 2001;363:17-26.

Matar C, Le Blanc JG, Martin L, Perdigon G. Biologically active peptides released in fermented milk; role and functions. In: Famworth ER. editor. Handbook of fermented functions foods. Functional foods and nutraceuticals series CRC Press: Boca Raton, USA; 2003. p. 177-201.

Korhonen H, Pihlanto-Leppala A. Milk-derived bioactive peptides: formation and prospects for health promotion. In C. short, JO Brien. ed. Handbook of Functional Dairy products. Functional Foods and Nutraceuticals Series 6.0, CRC press; Florida, USA: 2004. p. 109-24.

Demain AL, Somkuti GA, Hunter Levera JC, Rossmoore HW. Novel microbial products for medicine and Agriculture. Elsevier Sci Ltd; 1989. p. 161-9.

Valera HR, Gomes J, Lakshmi S, Gurujara Ra, Suryanarayan S, Kumar D. Lovastatin production by solid state fermentation using Aspergillus flavipes. Enzyme Microb Technol 2005;37:521-6.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265-75.

Cushman DW, Cheung HS. Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 1971;20:1637-48.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5.

Fomsgaard A, Conrad RS, Galanos C, Shand GH, Hoiby N. Comparative immunochemistry of Lipopolysaccharide from typable and poly agglutinable Pseudomonas aeruginosa strain isolated from patients with cystic fibrosis. J Clin Microbiol 1988;26:821-6.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.

Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980;284:555-6.

Mora L, Sentandreu MA, Koistinen KM, Fraser PD, Toldra F, Bramley PM. Naturally generated small peptides derived from myofibrillar proteins in Serrano dry-cured ham. J Agric Food Chem 2009a;57:3228-34.

Mora L, Sentandreu MA, Fraser PD, Toldra F, Bramley PM. Oligopeptides arising from the degradation of creatine kinasein spanish dry-cured ham. J Agric Food Chem 2009b;57:8982-8.

Sentandreu MA, Stoeva S, Aristoy MC, Laib K, Voelter W, Toldra E. Identification of small peptides generated in Spanish dry-cured ham. J Food Sci 2003;68:64-9.

Toldra F, Aristoy MC, Flores M. Contribution of muscle aminopeptidases to flavor development in dry-cured ham. Food Res Int 2000;33:181-5.

Sforza S, Pigazzani A, Motti M, Porta C, Virgili R, Galaverna G, et al. Oligopeptides and free amino acids in parmahams of known cathepsin B activity. Food Chem 2001;75:267-73.

Arihara K, Nakashima Y. Peptide inhibitors for angiotensin I converting enzyme from enzymatic hydrolyzates of porcine skeletal muscle proteins. Meat Sci 2001;57:319-24.

Katayama K, Tomatsu M, Kawahara S, Yamauchi K, Fuchu H, Kodama Y, et al. Inhibitory profile of nonapeptide derived from porcine troponin C against angiotensin I-converting enzyme. J Agric Food Chem 2004;52:771–5.

Escudero E, Sentandreu MA, Arihara K, Toldra F. Angiotensin I-converting enzyme inhibitory peptides generated from in vitro gastrointestinal digestion of pork meat. J Agric Food Chem 2010;58:2895−901.

Escudero E, Toldra F, Sentandreu MA, Nishimura H, Arihara K. Antihypertensive activity of peptides identified in the in vitro gastrointestinal digest of pork meat. Meat Sci 2012;91:382-4.

Ahmed H. Principle and reaction of protein extraction, purification, and characterization. USA: CRC press; 1959. p. 140-89.

Jung WK, Mendis E, Je JY, Park PJ, Son BW, Kim HC, et al. Angiotensin I converting enzyme inhibitory peptide from yellowfin sole (Limanda aspera) frame protein and its antihypertensive effect in spontaneously hypertensive rats. Food Chem 2006;94:26-32.

Jae JY, Lee KH, Lee MH, Ahn CB. Antioxidant and antihypertensive protein hydrolyzates produced from tuna liver by enzymatic hydrolysis. Food Res Int 2009;42:1266-72.

Wijesekara N. Dihydrofolate reductase mutations-associated megaloblastic anemia and cerebral folate deficiency. Clin Genet 2011;79:507-8.

Ko SC, Kang N, Kim EA, Kang CM, Lee SH, Kang SM, et al. A novel angiotensin I-converting enzyme (ACE) inhibitory peptide from a marine Chlorella ellipsoidea and its antihypertensive effect in spontaneously hypertensive rats. Process Biochem 2012;47:2005-2011.

Jumeri, Kim SM. Antioxidant and anticancer activities of enzymatic hydrolyzates of solitary tunicate (Styela clava). Food Sci Biotechnol 2011;20:1075–85.

Huang Y, Wang X, Wang H, Liu Y, Chen Y. Studies on the mechanism of action of anticancer peptides by modulation of hydrophobicity within a defined structural framework. Mol Cancer Ther 2011;10:416–26.

Cheung HS, Feng-Lai W, Ondetti MA, Sabo EF, Cushman DE. Binding of peptide substrates and inhibitors of angiotensin-converting-enzyme. J Biol Chem 1980;255:401-7.

Murray BA, FitzGerald RJ. Angiotensin converting enzyme inhibitory peptides derived from food proteins: biochemistry, bioactivity and production. Curr Pharm Des 2007;13:773-91.

Roberts PR, Burney JD, Black KW, Zaloga GP. Effect of chain length on the absorption of biologically active peptides from the gastrointestinal tract. Digestion 1999;60:332-7.

Castellano P, Aristoy MC, Sentandreu MA, Vignolo G, Toldra F. Peptides with angiotensin converting enzyme (ACE) inhibitory activity generated from porcine skeletal muscle proteins by the action of meat-borne Lactobacillus. J Proteomics 2013;89:183-90.

Parfrey PS. Inhibitors of the rennin-angiotensin system: proven benefits, unproven safety. Ann Intern Med 2008;148:76-7.

Lee SH, Qian ZJ, Kim SK. A novel angiotensin I converting enzyme inhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect in spontaneously hypertensive rats. Food Chem 2010;118:96–102.

Jang A, Jo C, Kang KS, Lee M. Antimicrobial and human cancer cell cytotoxic effect of the synthetic angiotensin-converting enzyme (ACE) inhibitory peptides. Food Chem 2008;107:327–36.

Wijesekara I, Kim SK. Angiotensin-I-converting enzyme (ACE) inhibitors from marine resources: prospects in the pharmaceutical industry. Mar Drugs 2010;8:1080–93.

Sharfi M, Schiffrin EL. Apoptosis in the vasculature of spontaneously hypertensic rats–effect, of an angiotensin converting enzyme inhibitor and calcium channel antagonist. Am J Hypertens 1998;11:1108-16.

Buemi M, Allegra A, Marino D, Marino MT, Medici MA, De Pasquale G, et al. Does captopril have a direct proapoptotic effect? Nephron 1999;81:99-101.

Daniel G, Larry B, Agnes V, Vivian T, Stuart G, Richard S, et al. Angiotensin-converting enzyme inhibition induces apoptosis in erythroid precursors and affects insulin like growth factor–I in post-transplantation erythrocytosis. J Am Soc Nephrol 2001;12:1958-64.

Wang Y, He H, Wang G, Wu H, Zhou B, Chen X, et al. Oyster (Crassostrea gigas) hydrolyzates produced on a plant scale has antitumor activity and immunostimulating effects in BALB/c Mice. Mar Drugs 2010;8:255–68.

Aleman A, Perez-Santin E, Bordenave-Juchereau S, Arnaudin I, Gomez-Guillen M, Montero P. Squid gelatin hydrolyzates with antihypertensive, anticancer and antioxidant activity. Food Res Int 2011;44:1044–51.

Ma A, Koka R, Burkett P. Diverse functions of IL-2, IL-15, and IL-7 in lymphoid homeostasis. Annu Rev Immunol 2006;24:657-79.

Medina RA, Goeger DE, Hills P, Mooberry SL, Huang N, Romero LI, et al. Coibamide A, a potent antiproliferative cyclic depsipeptide from the panamanian marine cyanobacterium Leptolyngbya sp. J Am Chem Soc 2008;130:6324–5.

Marquez BL, Watts KS, Yokochi A, Roberts MA, Verdier-Pinard P, Jimenez JI, et al. Structure and absolute stereochemistry of hectochlorin, a potent stimulator of actin assembly. J Nat Prod 2002;65:866–71.

Kalemkerian GP, Ou XL, Adil MR, Rosati R, Khoulani MM, Madan SK, et al. Activity of dolastatin 10 against small-cell lung cancer in vitro and in vivo: Induction of apoptosis and bcl-2 modification. Cancer Chemother Pharmacol 1999;43:507–15.

Mooberry SL, Leal RM, Tinley TL, Luesch H, Moore RE, Corbett TH. The molecular pharmacology of somatostatin 1: A new antimitotic dolastatin 10 analog. Int J Cancer 2003;104:512–21.

Costa DP, Huckstadt LA, Crocker DE, McDonald BI, Goebel ME, Fedak MA. Approaches to studying climatic change and its role on the habitat selection of Antarctic pinnipeds. Integr Comp Biol 2010;50:1018−30.

Costa M, Joao CR, Fernandes MH, Barros P, Vascibcekis V, Martins R. Marine cyanobacteria compound with anticancer properties: a review on the implication of apoptosis. Mar Drugs 2012;10:2181–207.

Landowski TH, Megli CJ, Nullmeyer KD, Lynch RM, Dorr RT. Mitochondrial-mediated disregulation of Ca2+is a critical determinant of Velcade (PS-341/bortezomib) cytotoxicity in myeloma cell lines. Cancer Res 2005;65:3828–36.

Mitsuoka K, Kato Y, Miyoshi S, Murakami Y, Hiraiwa M, Kubo Y, et al. Inhibition of oligopeptide transporter suppresses growth of human pancreatic cancer cells. Eur J Pharm Sci 2010;40:202-8.

Brandsch M. Transport of drugs by proton-coupled peptide transporters: pearls and pitfalls. Expert Opin Drug Metab Toxicol 2009;5:887-905.

Ahuja D, Geiger A, Ramanjulu J, Vera M, Sir Deshpande B, Pfizenmayer A, et al. Inhibition of protein synthesis by didemnins: Cell potency and SAR. J Med Chem 2000;43:4212–8.

Mayer AM, Gustafson KR. Marine pharmacology in 2000: antitumor and cytotoxic compounds. Int J Cancer 2003;105:291–9.

Nakazawa H, Kitano K, Cioca D, Ishikawa M, Ueno M, Ishida F, et al. Induction of polyploidization by jaspamide in HL-60 cells. Acta Haematol 2000;104:65–71.

Panda D, Ananthnarayan V, Larson G, Shih C, Jordan M, Wilson L. Interaction of the antitumor compound cryptophycin-52 with tubulin. Biochemistry 2000;39:14121–7.

Prasanna PG, Stone HB, Wong RS, Capala J, Bernhard EJ, Vikram B, et al. Normal tissue protection for improving radiotherapy: where are the Gaps? Transl Cancer Res 2012;1:35-48.



How to Cite

Ebrahimi, L., K. A. A. Nawaz, K. G. Kiran, S. D. Nisshanthini, M. Palaniswamy, and J. Angayarkanni. “PRODUCTION AND PURIFICATION OF ANGIOTENSIN-CONVERTING ENZYME INHIBITOR BY SELECTED BACTERIAL STRAIN FOR CANCER THERAPY”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 8, no. 2, Feb. 2016, pp. 214-20,



Original Article(s)

Most read articles by the same author(s)