Applied Biochemistry and Biotechnology

, Volume 177, Issue 1, pp 137–147 | Cite as

Characterization of an Antibacterial Compound, 2-Hydroxyl Indole-3-Propanamide, Produced by Lactic Acid Bacteria Isolated from Fermented Batter

  • Kadirvelu JeevaratnamEmail author
  • Venkatasubramanian Vidhyasagar
  • Perumal Jayaprabha Agaliya
  • Appukuttan Saraniya
  • Muthukandan Umaiyaparvathy


Lactic acid bacteria are known to produce numerous antimicrobial compounds that are active against various pathogens. Here, we have purified and characterized a novel low-molecular-weight (LMW) antimicrobial compound produced by Lactobacillus and Pediococcus isolated from fermented idly and uttapam batter. The LMW compound was extracted from cell-free supernatant using ice-cold acetone, purified by gel permeation and hydrophobic interaction chromatography. It exhibited antimicrobial activity against Gram-positive and Gram-negative pathogenic bacteria sparing the probiotic strains like Lactobacillus rhamnosus. The molecular weight of the LMW compound was identified as 204 Da using LC-MS-ESI. In addition, the structure of the compound was predicted using spectroscopic methods like FTIR and NMR and identified as 2-hydroxyl indole-3-propanamide. The LMW compound was differentiated from its related compound, tryptophan, by Salkowski reaction and thin-layer chromatography. This novel LMW compound, 2-hydroxyl indole-3-propanamide, may have an effective application as an antibiotic which can spare prevailing probiotic organisms but target only the pathogenic strains.


Lactobacillus Pediococcus Antibacterial NMR FTIR 2-Hydroxyl indole-3-propanamide 



The authors are thankful to ER&IPR, Defense Research & Development Organization, New Delhi, for funding this project and Defence R&D Establishment, Gwalior, for providing facilities for structural elucidation of this compound. Ms. Jayaprabha Agaliya is thankful for providing fellowship in this project, while Ms. Saraniya and Mr. Vidhyasagar are thankful to University Grants Commission, New Delhi, for providing fellowship.

Supplementary material

12010_2015_1733_MOESM1_ESM.docx (278 kb)
ESM 1 (DOCX 277 kb)


  1. 1.
    Smid, E. J., & Kleerebezem, M. (2014). Production of aroma compounds in lactic fermentations. Annual Review of Food Science and Technology, 5, 313–326.CrossRefGoogle Scholar
  2. 2.
    Broberg, A., Jacobsson, K., Strom, K., & Schnurer, J. (2007). Metabolite profiles of lactic acid bacteria in grass silage. Applied and Environmental Microbiology, 73, 5547–5552.CrossRefGoogle Scholar
  3. 3.
    Duckstein, S. M., Lorenz, P., & Stintzing, F. C. (2012). Conversion of phenolic constituents in aqueous Hamamelis virginiana leaf extracts during fermentation. Phytochemical Analysis, 23, 588–597.CrossRefGoogle Scholar
  4. 4.
    Yang, Z., Suomalainen, T., Mayra-Makinen, A., & Huttunen, E. (1997). Antimicrobial activity of 2-pyrrolidone-5-carboxylic acid produced by lactic acid bacteria. Journal of Food Protection, 60, 786–790.Google Scholar
  5. 5.
    Niku-Paavalo, M. L., Laitila, A., Mattila-Sandholm, T., & Haikara, A. (1999). New type of antimicrobial compounds produced by Lactobacillus plantarum. Journal of Applied Microbiology, 86, 29–35.CrossRefGoogle Scholar
  6. 6.
    Knockaert, D., Raes, K., Wille, C., Struijsa, K., & Van Camp, J. (2012). Metabolism of ferulic acid during growth of Lactobacillus plantarum and Lactobacillus collinoides. Journal of the Science of Food and Agriculture, 92, 2291–2296.CrossRefGoogle Scholar
  7. 7.
    Rodriguez, N., Salgado, J. M., Cortes, S., & Domínguez, J. M. (2012). Antimicrobial activity of D-3-phenyllactic acid produced by fed-batch process against Salmonella enterica. Food Control, 25, 274–284.CrossRefGoogle Scholar
  8. 8.
    Sjogren, J., Magnusson, J., Broberg, A., Schnurer, J., & Kenne, L. (2003). Antifungal 3-hydroxy fatty acids from Lactobacillus plantarum MiLAB 14. Applied and Environmental Microbiology, 69, 7554–7557.CrossRefGoogle Scholar
  9. 9.
    Chung, T. C., Axelsson, L., Lindgren, S. E., & Dobrogosz, W. J. (1989). In vitro studies on reuterin synthesis by Lactobacillus reuteri. Microbial Ecology in Health and Disease, 2, 137–144.CrossRefGoogle Scholar
  10. 10.
    El-Ziney, M. G., Debevere, J. M., & Jakobsen, M. (2000). In A. S. Naidu (Ed.), Natural food antimicrobial systems (pp. 567–588). Florida: CRC Press LLC.Google Scholar
  11. 11.
    Ganzle, M. G., Zhang, C., Monang, B. S., Lee, V., & Schwab, C. (2009). Novel metabolites from cereal-associated lactobacilli—novel functionalities for cereal products? Food Microbiology, 26, 712–719.CrossRefGoogle Scholar
  12. 12.
    Jeevaratnam, K., Jamuna, M., & Bawa, A. S. (2005). Biological preservation of foods-bacteriocins of lactic acid bacteria. Indian Journal of Biotechnology, 4, 446–454.Google Scholar
  13. 13.
    Vidhyasagar, V., & Jeevaratnam, K. (2012). Isolation and characterization of Pediococcus pentosaceus from idly batter: a traditional South Indian fermented food source. Biosciences, Biotechnology Research Asia, 9, 427–431.CrossRefGoogle Scholar
  14. 14.
    Agaliya, P. J., & Jeevaratnam, K. (2013). Molecular characterization of lactobacilli isolated from fermented idli batter. Brazilian Journal of Microbiology, 44, 1199–1206.CrossRefGoogle Scholar
  15. 15.
    Saraniya, A., & Jeevaratnam, K. (2012). Molecular characterization of bacteriocinogenic Lactobacillus species isolated from fermented Uttapam batter. Biosciences, Biotechnology Research Asia, 9, 417–421.CrossRefGoogle Scholar
  16. 16.
    Parente, E., Brienza, C., Moles, M., & Ricciardi, A. A. (1995). A comparison of methods for the measurement of bacteriocin activity. Journal of Microbiological Methods, 22, 95–108.CrossRefGoogle Scholar
  17. 17.
    Ehmann, A. (1977). The van Urk-Salkowski reagent—a sensitive and specific chromogenic reagent for silica gel thin-layer chromatographic detection and identification of indole derivatives. Journal of Chromatography A, 1977, 267–276.CrossRefGoogle Scholar
  18. 18.
    Stoyanova, L. G., Ustyugova, E. A., & Netrusov, A. I. (2012). Antibacterial metabolites of lactic acid bacteria: their diversity and properties. Applied Biochemistry and Microbiology, 48, 229–243.CrossRefGoogle Scholar
  19. 19.
    Agaliya, P. J., & Jeevaratnam, K. (2012). Screening of Lactobacillus plantarum isolated from fermented idli batter for probiotic properties. African Journal of Biotechnology, 11, 12856–12864.Google Scholar
  20. 20.
    Saraniya, A. and Jeevaratnam, K. (2014) In vitro probiotic evaluation of phytase producing Lactobacillus species isolated from Uttapam batter and their application in soy milk fermentation. J Food Sci Technol, 1-10.Google Scholar
  21. 21.
    Vidhyasagar, V., & Jeevaratnam, K. (2013). Evaluation of Pediococcus pentosaceus strains isolated from idly batter for probiotic properties in vitro. Journal of Functional Foods, 5, 235–243.CrossRefGoogle Scholar
  22. 22.
    Agaliya, P. J., & Jeevaratnam, K. (2013). Characterisation of the bacteriocins produced by two probiotic Lactobacillus isolates from idli batter. Annals of Microbiology, 63, 1525–1535.CrossRefGoogle Scholar
  23. 23.
    Saraniya, A., & Jeevaratnam, K. (2014). Purification and mode of action of antilisterial bacteriocins produced by Lactobacillus pentosus SJ65 isolated from Uttapam batter. Journal of Food Biochemistry, 38, 612–619.CrossRefGoogle Scholar
  24. 24.
    Vidhyasagar, V., & Jeevaratnam, K. (2013). Bacteriocin activity against various pathogens produced by Pediococcus pentosaceus VJ13 isolated from Idly batter. Biomedical Chromatography, 27, 1497–1502.CrossRefGoogle Scholar
  25. 25.
    Silva, M., Jacobus, N. V., Deneke, C., & Gorbachl, S. L. (1987). Antimicrobial substance from a human Lactobacillus strain. Antimicrobial Agents and Chemotherapy, 31, 1231–1233.CrossRefGoogle Scholar
  26. 26.
    Breitmaier, E. (2002). Structure elucidation by NMR in inorganic chemistry: a practical guide (3rd ed.). England: Wiley Ltd.CrossRefGoogle Scholar
  27. 27.
    Olgen, S., Altanlar, N., Karatayli, E., & Bozdayi, M. (2008). Antimicrobial and antiviral screening of novel indole carboxamide and propanamide derivatives. Zeitschrift Naturforschung C, 63, 189–195.Google Scholar
  28. 28.
    Rohini, R., Reddy, P. M., Shanker, K., Kanthaiah, K., Ravinder, V., & Hu, A. (2011). Synthesis of mono, bis-2-(2-arylideneaminophenyl) indole azomethines as potential antimicrobial agents. Archives of Pharmacal Research, 34, 1077–1084.CrossRefGoogle Scholar
  29. 29.
    Lee, J. H., & Lee, J. (2010). Indole as an intercellular signal in microbial community. FEMS Microbiology Review, 34, 426–444.CrossRefGoogle Scholar
  30. 30.
    Sajid, I., Shaaban, K. A., & Hasnain, S. (2011). Identification, isolation and optimization of antifungal metabolites from the Streptomyces malachitofuscus ctf9. Brazilian Journal of Microbiology, 42, 592–604.CrossRefGoogle Scholar
  31. 31.
    Carbonnelle, D., Duflos, M., Marchand, P., Chauvet, C., Petit, J.-Y., & Lang, F. (2009). A novel indole-3-propanamide exerts its immunosuppressive activity by inhibiting JAK3 in T Cells. Journal of Pharmacol and Experimental Therapeutics, 331, 710–716.CrossRefGoogle Scholar
  32. 32.
    Wang, H., Yan, Y., Wang, J., Zhang, H., & Qi, W. (2012). Production and characterization of antifungal compounds produced by Lactobacillus plantarum IMAU10014. PLoS ONE, 7, e29452. doi: 10.1371/journal.pone.0029452.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Kadirvelu Jeevaratnam
    • 1
    Email author
  • Venkatasubramanian Vidhyasagar
    • 1
  • Perumal Jayaprabha Agaliya
    • 1
  • Appukuttan Saraniya
    • 1
  • Muthukandan Umaiyaparvathy
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyPondicherry UniversityKalapetIndia

Personalised recommendations