Current Microbiology

, Volume 73, Issue 6, pp 885–896 | Cite as

Antibacterial Activity of Polyphenolic Fraction of Kombucha Against Enteric Bacterial Pathogens

  • Debanjana Bhattacharya
  • Semantee Bhattacharya
  • Madhu Manti Patra
  • Somnath Chakravorty
  • Soumyadev Sarkar
  • Writachit Chakraborty
  • Hemanta Koley
  • Ratan GachhuiEmail author


The emergence of multi-drug-resistant enteric pathogens has prompted the scientist community to explore the therapeutic potentials of traditional foods and beverages. The present study was undertaken to investigate the efficacy of Kombucha, a fermented beverage of sugared black tea, against enterotoxigenic Escherichia coli, Vibrio cholerae, Shigella flexneri and Salmonella Typhimurium followed by the identification of the antibacterial components present in Kombucha. The antibacterial activity was evaluated by determining the inhibition zone diameter, minimal inhibitory concentration and minimal bactericidal concentration. Kombucha fermented for 14 days showed maximum activity against the bacterial strains. Its ethyl acetate extract was found to be the most effective upon sequential solvent extraction of the 14-day Kombucha. This potent ethyl acetate extract was then subjected to thin layer chromatography for further purification of antibacterial ingredients which led to the isolation of an active polyphenolic fraction. Catechin and isorhamnetin were detected as the major antibacterial compounds present in this polyphenolic fraction of Kombucha by High Performance Liquid Chromatography. Catechin, one of the primary antibacterial polyphenols in tea was also found to be present in Kombucha. But isorhamnetin is not reported to be present in tea, which may thereby suggest the role of fermentation process of black tea for its production in Kombucha. To the best of our knowledge, this is the first report on the presence of isorhamnetin in Kombucha. The overall study suggests that Kombucha can be used as a potent antibacterial agent against entero-pathogenic bacterial infections, which mainly is attributed to its polyphenolic content.


High Performance Liquid Chromatography Minimal Inhibitory Concentration Catechin Ethyl Acetate Extract Minimal Bactericidal Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Dr. Joydeep Mukherjee, UGC Associate Professor and Dr. Sayani Mitra, Mr. Kaushik Biswas and Mr. Sayak Bhattacharya of School of Environmental Studies, Jadavpur University, Kolkata for HPLC analysis and Prof. Mahammad Ali, Department of Chemistry, Jadavpur University, Kolkata for ESI–MS study. We also thank Dr. Saubhik Haldar of Department of Chemistry, Jadavpur University, Kolkata for analysis of the sample and Dr. Dhrubajyoti Nag, Dr. Ritam Sinha and Ms. Priyadarshini Mukherjee of National Institute of Cholera and Enteric Diseases, Kolkata for efficient assistance in the study. This work has been financially assisted by the Department of Science and Technology, Government of India.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Arakawa H, Maeda M, Okubo S, Shimamura T (2004) Role of hydrogen peroxide in bactericidal action of catechin. Biol Pharm Bull 27:277–281CrossRefPubMedGoogle Scholar
  2. 2.
    Bag A, Bhattacharyya SK, Pal NK, Chattopadhyay RR (2012) In vitro antibacterial potential of Eugenia jambolana seed extracts against multi-drug-resistant human bacterial pathogens. Microbiol Res 167:352–357CrossRefPubMedGoogle Scholar
  3. 3.
    Barman S, Chatterjee S, Chowdhury G, Ramamurthy T, Niyogi SK, Kumar R, Koley H (2010) Plasmid-mediated streptomycin and sulfamethoxazole resistance in Shigella flexneri 3a. Int J Antimicrob Ag 36:348–351CrossRefGoogle Scholar
  4. 4.
    Battikh H, Chaieb K, Bakhrouf A, Ammar E (2013) Antibacterial and antifungal activities of black and green Kombucha teas. J Food Biochem 27:231–236CrossRefGoogle Scholar
  5. 5.
    Bhattacharya S, Gachhui R, Sil PC (2011) Hepatoprotective properties of Kombucha tea against TBHP-induced oxidative stress via suppression of mitochondria dependent apoptosis. Pathophysiology 18:221–234CrossRefPubMedGoogle Scholar
  6. 6.
    Bhattacharya S, Gachhui R, Sil PC (2013) Effect of Kombucha, a fermented black tea in attenuating oxidative stress mediated tissue damage in alloxan induced diabetic rats. Food Chem Toxicol 60:328–340CrossRefPubMedGoogle Scholar
  7. 7.
    Bhattacharya S, Manna P, Gachhui R, Sil PC (2011) Protective effect of Kombucha tea against tertiary butyl hydroperoxide induced cytotoxicity and cell death in murine hepatocytes. Indian J Exp Biol 49:511–524PubMedGoogle Scholar
  8. 8.
    Chakravorty S, Bhattacharya S, Chatzinotas A, Chakraborty W, Bhattacharya D, Gachhui R (2016) Kombucha tea fermentation: microbial and biochemical dynamics. Int J Food Microbiol 220:63–72CrossRefPubMedGoogle Scholar
  9. 9.
    Chang TKH, Chen J, Yeung EYH (2006) Effect of Ginkgo biloba extract on procarcinogen-bioactivating human CYP1 enzymes: identification of isorhamnetin, kaempferol, and quercetin as potent inhibitors of CYP1B1. Toxicol Appl Pharm 213:18–26CrossRefGoogle Scholar
  10. 10.
    Chattopadhyay RR, Bhattacharyya SK, Medda C, Chanda S, Bag A (2009) A comparative evaluation of antibacterial potential of some plants used in Indian traditional medicine for the treatment of microbial infections. Braz Arch Biol Techn 52:1123–1128CrossRefGoogle Scholar
  11. 11.
    Cushnie TPT, Lamb AJ (2011) Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Ag 38:99–107CrossRefGoogle Scholar
  12. 12.
    Daglia M (2012) Polyphenols as antimicrobial agents. Curr Opin Biotech 23:174–181CrossRefPubMedGoogle Scholar
  13. 13.
    Dufresne C, Farnworth E (2000) Tea, Kombucha, and health: a review. Food Res Int 33:409–421CrossRefGoogle Scholar
  14. 14.
    Dutta D, Gachhui R (2006) Novel nitrogen-fixing Acetobacter nitrogenifigens sp. nov., isolated from Kombucha tea. Int J Syst Evol Microbiol 56:1899–1903CrossRefPubMedGoogle Scholar
  15. 15.
    Dutta D, Gachhui R (2007) Nitrogen-fixing and cellulose-producing Gluconacetobacter kombuchae sp. nov., isolated from Kombucha tea. Int J Syst Evol Microbiol 57:353–357CrossRefPubMedGoogle Scholar
  16. 16.
    Greenwalt CJ, Ledford RA, Steinkraus KH (1998) Determination and characterization of the antimicrobial activity of the fermented tea Kombucha. Food Sci Technol-LEB 31:291–296CrossRefGoogle Scholar
  17. 17.
    Jayabalan R, Chen P-N, Hsieh Y-S, Prabhakaran K, Pitchai P, Marimuthu S, Thangaraj P, Swaminathan K, Yun SE (2011) Effect of solvent fractions of Kombucha tea on viability and invasiveness of cancer cells—characterization of dimethyl 2-(2-hydroxy-2 methoxypropylidine) malonate and vitexin. Indian J Biotechnol 10:75–82Google Scholar
  18. 18.
    Jayabalan R, Malbaša RV, Lončar ES, Vitas JS, Sathishkumar M (2014) A review on Kombucha tea—microbiology, composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr Rev Food Sci Food Saf 13:538–550CrossRefGoogle Scholar
  19. 19.
    Mehri D, Monsef-Esfahani HR, Gharibzadeh S, Jafari K, Faghihi M (2008) Effects of black tea extract and its thearubigins on whole gut transit time in mice: involvement of 5-HT3 receptors. Jundishapur J Nat Pharm Prod 3:39–44Google Scholar
  20. 20.
    NCCLS: National Committee for Clinical Laboratory Standards (1997) Performance standards for antimicrobial disk susceptibility test, 6th edn. NCCLS, WayneGoogle Scholar
  21. 21.
    Nenaah G (2013) Antimicrobial activity of Calotropis procera Ait. (Asclepiadaceae) and isolation of four flavonoid glycosides as the active constituents. World J Microbiol Biotechnol 29:1255–1262CrossRefPubMedGoogle Scholar
  22. 22.
    Rojas A, Hernandez L, Pereda-Miranda R, Mata R (1992) Screening for antimicrobial activity of crude drug extracts and pure natural products from Mexican medicinal plants. J Ethnopharmacol 35:275–283CrossRefPubMedGoogle Scholar
  23. 23.
    Ross ZM, OʼGara EA, Hill DJ, Sleightholme HV, Maslin DJ (2001) Antimicrobial properties of garlic oil against human enteric bacteria: evaluation of methodologies and comparisons with garlic oil sulfides and garlic powder. Appl Environ Microbiol 67:475–480CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Sánchez-Rabaneda F, Jáuregui O, Casals I, Andrés-Lacueva C, Izquierdo-Pulido M, Lamuela-Raventós RM (2003) Liquid chromatographic/electrospray ionization tandem mass spectrometric study of the phenolic composition of cocoa (Theobroma cacao). J Mass Spectrom 38:35–42CrossRefPubMedGoogle Scholar
  25. 25.
    Sreeramulu G, Zhu Y, Knol W (2000) Kombucha fermentation and its antimicrobial activity. J Agric Food Chem 48:2589–2594CrossRefPubMedGoogle Scholar
  26. 26.
    Srihari T, Karthikesan K, Ashokkumar N, Satyanarayana U (2013) Antihyperglycaemic efficacy of kombucha in streptozotocin-induced rats. J Funct Foods 5:1794–1802CrossRefGoogle Scholar
  27. 27.
    Taguri T, Tanaka T, Kouno I (2006) Antibacterial spectrum of plant polyphenols and extracts depending upon hydroxyphenyl structure. Biol Pharm Bull 29:2226–2235CrossRefPubMedGoogle Scholar
  28. 28.
    Teng B-S, Lua Y-H, Wang Z-T, Tao X-Y, Wei D-Z (2006) In vitro anti-tumor activity of isorhamnetin isolated from Hippophae rhamnoides L. against BEL-7402 cells. Pharmacol Res 54:186–194CrossRefGoogle Scholar
  29. 29.
    Van der Watt E, Pretorius JC (2001) Purification and identification of active antibacterial components in Carpobrotus edulis L. J Ethnopharmacol 76:87–91CrossRefPubMedGoogle Scholar
  30. 30.
    Veluri R, Weir TL, Bais HP, Stermitz FR, Vivanco JM (2004) Phytotoxic and antimicrobial activities of catechin derivatives. J Agric Food Chem 52:1077–1082CrossRefPubMedGoogle Scholar
  31. 31.
    Yin H, Deng Y, Wang H, Liu W, Zhuang X, Chu W (2015) Tea polyphenols as an antivirulence compound disrupt quorum-sensing regulated pathogenicity of Pseudomonas aeruginosa. Sci Rep. doi: 10.1038/srep16158 Google Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Debanjana Bhattacharya
    • 1
  • Semantee Bhattacharya
    • 1
  • Madhu Manti Patra
    • 1
    • 3
  • Somnath Chakravorty
    • 1
    • 4
  • Soumyadev Sarkar
    • 1
  • Writachit Chakraborty
    • 1
  • Hemanta Koley
    • 2
  • Ratan Gachhui
    • 1
    Email author
  1. 1.Department of Life Science & BiotechnologyJadavpur UniversityKolkataIndia
  2. 2.Division of BacteriologyNational Institute of Cholera and Enteric Diseases (NICED)KolkataIndia
  3. 3.Department of MicrobiologyBose InstituteKolkataIndia
  4. 4.Department of Biochemistry and Molecular BiophysicsKansas State UniversityManhattanUSA

Personalised recommendations