Enzymatic Activity of Bromelain Isolated Pineapple (Ananas comosus) Hump and Its Antibacterial Effect on Enterococcus faecalis

Dewi Liliany, Armelia Sari Widyarman, Erni Erfan, Janti Sudiono, Melanie Sadono Djamil


Background: Enterococcus faecalis is the frequent cause of oral infections, such as periodontitis, infected root canals, and peri-radicular abscesses. Pineapple (Ananas comosus) fruit contains bromelain, one of proteolytic enzymes associated with several health benefits. Bromelain has been shown to promote healthy digestion, stimulate the immune system, improve cardiovascular conditions, and accelerate wound healing. Bromelain compounds possess anti-inflammatory and anticancer properties and exhibit antibacterial activity. Objective: To analyze the enzymatic activity of bromelain extracted from pineapple hump and investigate the antibacterial effect of bromelain against E. faecalis. Methods: Pineapple hump was dried and extracted with maceration technique. Further purification was obtained by ammonium sulfate fractionation, dialysis and ion exchange chromatography. Minimum inhibitory concentration (MIC) tests using diffusion and dilution techniques tested the antibacterial activity of the bromelain extract on E. faecalis. A one-way analysis of variance (ANOVA) test analyzed the significance of the differences in the E. faecalis inhibition zones after treatment with a range of bromelain extract concentrations. Differences were considered statistically significant if p < 0.05. Results: The specific activity of bromelain in the crude extract was 62.89 U/mg. Furthermore, bromelain activity using ammonium sulfate fractionation was 50.99 U/mg, dialysis was 54.59 U/mg, and ion exchange chromatography was 152.38 U/mg. The bromelain extract showed effective inhibitory and bactericidal activity against E. faecalis. The results of the inhibition test using a bromelain extract purified by ion exchange  chromatography demonstrated that a concentration as small as 12.5% was effective in inhibiting the growth of E. faecalis (p < 0.05). Conclusion: The highest enzymatic activity of bromelain was found after purification with ion exchange chromatography. Bromelain exerted an antibacterial effect against a potent endodontic pathogen, but further studies are needed to explore this effect.

Full Text:


Article Metrics

Abstract views : 557| PDF views : 0


Walton RE, Rivera EM. Cleaning and Shaping. In: Walton RE, Torabinejad M, editors Principles and Practice of Endodontic. 3rd ed. Philadelphia: WB Saunders: 2002. p.206-38.

Evan M, Davies JK, Sundqvist G, Fidgor D. Mechanisms involved in the resistance of the Enterococcus faecalis to calcium hydroxide. Int Endod. 2002: 35(3) : 221-8.

Portenier I, Waltimo T M, Haapsalo M. Enterococcus faecalis the root canal survival and star in post treatment disease. Endod Topics. 2003: 6(1) : 135-39. DOI: https://doi.org/10.1111/j.1601-1546.2003.00040.x

Fisher K, Philips C. The ecology, epidemiology and virulence of Enterococcus. Microbiology. 2009: 155(Pt 6) : 1749-55. DOI: 10.1099/mic.0.026385-0

Love RM. Enterococcus faecalis a mechanism for its role in endodontic failure. Int Endod J. 2001; 34(5) : 399-405.

Sundqvist G, Fidgor D. Life as an endodontic pathogen: ecological difference between untreated and filled root canal. Endod Topics. 2003; 6(1): 3-28. DOI: https://doi.org/10.1111/j.1601-1546.2003.00054.x

Madigan MT, Martinko JM, Bender KS, Buckley DH, Stahl DA, Brock TD. Brock Biology of Micro organisms. 11th ed. New Jersey: Pearson Prentice Hall; 2006. p. 75-77

de la Maza LM, Pezzlo MT, Shigei JT, Tan GL, Peterson EM. Color Atlas of Medical Bacteriology. 2nd ed. Washington DC: American Society for Microbiology Press, 2013.

Signoretto C, Lleò MM, Tafi MC, Canepari P. Cell wall chemical composition of Enterococcus faecalis in the viable but nonculturable state. Appl Environ Microbiol.2000; 66(5):1953-9.

Suchitra U, Kundabala M. Enterococcus faecalis: An endodontic pathogen. Endodontology. 2002;18(3) 11-3.

Mundy LM, Sahm DF, Gilmore M. Relationships between Enterococcal virulence and antimicrobial resistance. Clin Microbiol Rev. 2000; 13(4): 513 – 522.

Kandaswamy D, Venkateshbabu N. Root canal irrigants. J. Conserv Dent. 2010; 13(4): 256-264. DOI: 10.4103/0972-0707.73378

Nanas Subang, Diminati Dunia. 15 April 2010. Diakses 15 Oktober 2013. Available from: http://bisnisukm.com/nanas-subang-diminati-dunia-html/.

Kumaunang. Aktivitas Enzim Bromelain dan Ekstrak Kulit Nanas (Ananas comosus). Universitas Sam Ratulangi. Manado. 2010.

Gautam SS, Mishra SK, Dash V, Goyal AK, Rath G. Cooperative study or extraction, purification and Estimation of bromelain from stem and fruit from pineapple plant. Thai J. Pharm Sci. 2010; 34: 67-76.

Kelly GS. Bromelain : a literature review and discussion of its therapeutic application. Alt. Med. Rev. 1996; 1(4): 243-5.

Drauz K, Waldmann H. Enzym Catalysis in Organic Synthesis : A Comprehensive Handbook Vol.1. Weinheim: Wiley-VCH; p.346. DOI: https://doi.org/10.1002/9783527619429.fmatter

Taussig SJ, Batkin S. Bromelain, the enzyme complex of pineapple (Ananas comosus) and its clinical applications, an update. J. Ethnopharmacol. 1988, 22(2): 191-203. DOI: https://doi.org/10.1016/0378-8741(88)90127-4

Lotz-Winter H. On the pharmacology of Bromelain: an update with special regard to animal studies on dose-dependent effects. Planta Med. 1990; 56(3): 249-53. DOI: 10.1055/s-2006-960949

Maurer H.R, Eckert K. Bromelain in der komplemention Tumor terapy. Onkol/J Oncol. 1999; 31: 66-7.

Brakebusch M, Wintergerst U, Petropoulou T, et al. Bromelain is an accelerator of phagocytosis, respiratory burst and killing of Candida albicans by human granulocytes and monocytes. Eur J Med Res. 2001; 6(5):193-200.

Praveen NC, Rajesh A, Madan M, Chaurasia VR, Hiremath NV, Sharma AM. In vitro evaluation of antibacterial efficacy of pineapple extract (bromelain) on periodontal pathogens. J Int Oral Health. 2014; 6(5):96-8.

Scopes R.K. Protein Purification Principles and Practice. 2nd ed. New York: Springer Verlag; 1987. p.44.

Md. Farid Hossain, Shaheen Akhtar, Mustafa Anwar. Nutritional value and medicinal benefits of pineapple. International Journal of Nutrition and Food Sciences. 2015; 4(1): p. 84-88. DOI: 10.11648/j.ijnfs.20150401.22

Belton PS, Sorensen H, Sorensen S, Bjergegaard C, Michaelson S. High-performance capillary electrophoresis. In: Sorensen H, Sorensen S, Bjergegaard C, Michelson S, editors. Chromatography and Capillary Electrophoresis in Food Analysis. Cambridge: RSC; 1999. p. 223-37

Kaur T, Kaur A, Grewal RK. Kinetics studies with fruit bromelain (Ananas comosus) in the presence of cysteine and divalent ions. J Food Sci Technol. 2015; 52(9): 5954–60.

Herdyastuti N. Isolasi dan karakteristik ekstrak kasar enzim bromelain dari batang nanas. Journal of Biological Researches. 2006; 12: 75-7.

Caesarita DP. Pengaruh ekstrak buah nanas (Ananascosmosus) 100% terhadap S. aureus penyakit pioderma. [Unpublished Undergraduate Thesis]. Semarang: Faculty of Medicine Diponegoro University; 2011.

Stepek G, Lowe AE, Buttle DJ, et al. In vitro and in vivo anthelmintic efficacy of plantcysteine proteinases against the rodentgastrointestinal nematode, Trichurismuris. Parasitology. 2006; 132(Pt5): 681-9. DOI : 10.1017/S003118200500973X

Stepek G, Buttle DJ, Duce IR, Lowe A, Behnke JM. Assessment of the anthelmintic effect of natural plant cysteine proteinases against the gastrointestinal nematode, Heligmosomoidespolygyrus, in vitro. Parasitology 2005; 130(Pt 2): 203-211.

Mynott TL, Guandalini S, Raimondi F, Fasano A. Bromelain prevents secretion caused by Vibrio cholerae and Escherichia coli enterotoxins in rabbit ileum in vitro. Gastroenterology 1997;113 (1):175-184.

Winarsih S, Asmaningsih E, Nina A. Efektivitas Ekstrak Etanol Kulit Anggur Bali (vitisvinifera L. var. Alphonse lavallee) Sebagai Antibakteri Terhadap Escherichia coli Secara in vitro. [Unpulished Thesis] Malang: Faculty of Medicine Brawijaya University; 2014. Available from : http:/repository.ub.ac.id/id/eprint/26994

Widyarman AS, Widjaja SB, Idrus E. Strawberry extract's effects on Enterococcus faecalis and Porphyromonas gingivalis biofilms in vitro. Sci Dent J. 2017; 1(1):1–5. DOI: http://dx.doi.org/10.26912/sdj.v1i1

Ali AA, Milala MA, Gulani IA. Antimicrobial effects of crude bromelain extracted from pineapple fruit (Ananas comosus (Linn.) Merr.). Advances in Biochemistry. 2015; 3(1) :1-4. DOI: 10.11648/j.ab.20150301.11

Zainol, N.A., Voo S.C, Sarmidi, M.R., and Aziz, R.A. Profilling of Centella asiatica (L) urban extract. MJAS. 2008; 12(2): 322-27.

Heinicke R. Stem Bromelain a New Protease Preparation from Pineapple Plants. Econ Bot 2007. p.225-34.

Tussig SJ, Batkin S. Bromelain, the enzyme complex of pineapple (Ananas cosmosus) and its clinical applications, an update. J Ethnopharmacol. 1998; 27: 191-3.

Lotz-Winter H. On the pharmacology of Bromelain: an update with special regard to animal studies on dose-dependent effects. Planta Med. 1990; 56(3): 249-51.DOI: 10.1055/s-2006-960949

Kelly GS. Bromelain : a literature review and discussion of its therapeutic application. Alt. Med. Rev 1996, 1(4): 243-45.

Maurer HR, Eckert K. Bromelain in der komplemention Tumor terapy. Onkol/J Oncol. 1999; 31: 66-7.

DOI: http://dx.doi.org/10.26912/sdj.v2i2.2540


  • There are currently no refbacks.

Copyright (c) 2018

Copyright of Scientific Dental Journal. (e-ISSN: 2541–321X, p-ISSN: 2580–6548)

Scientific Dental Journal is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Creative Commons License