Amplification of Vacuole-targeting Fungicidal Activity of Antibacterial Antibiotic Polymyxin B by Allicin, an Allyl Sulfur Compound from Garlic


A cationic antibacterial peptide, polymyxin B (PMB), was evaluated as an antifungal antibiotic against various yeasts and filamentous fungi when used in combination with allicin, an allyl sulfur compound from garlic. Allicin was not lethal but could markedly amplify the fungicidal activity of PMB, which was weakly detected with the increase in the plasma membrane permeability in Saccharomyces cerevisiae. Their combined actions caused a dynamic structural damage to the yeast vacuole as judged by the disappearance of its swollen spherical architecture. The vacuole-targeting activity of PMB was similarly amplified in medium with t-butyl hydroperoxide as a substitute for the action of allicin. These findings suggest that the allicin-mediated lipoperoxide production in fungal plasma membrane is the cause of the enhancement in the cellular uptake of PMB as well as its action against the vacuole.


  1. 1

    Storm DR, Rosenthal KS, Swanson PE . Polymyxin and related peptide antibiotics. Annu Rev Biochem 46: 723–763 ( 1977)

    CAS  Article  Google Scholar 

  2. 2

    Srimal S, Surolia N, Balasubramanian S, Surolia A . Titration calorimetric studies to elucidate the specificity of the interactions of polymyxin B with lipopolysaccharides and lipid A. Biochem J 315: 679–686 ( 1996)

    CAS  Article  Google Scholar 

  3. 3

    Tsang JC, Weber DA, Brown DA . Evidences for complex formation between polymyxin B and lipopolysaccharides from Serratia marcescens. J Antibiot 29: 735–742 ( 1976)

    CAS  Article  Google Scholar 

  4. 4

    Hancock REW, Chapple D . Peptide antibiotics. Antimicrob Agents Chemother 43: 1317–1323 ( 1999)

    CAS  Article  Google Scholar 

  5. 5

    Teuber M, Bader J . Action of polymyxin B on bacterial membranes. Binding capacities for polymyxin B of inner and outer membranes isolated from Salmonella typhimurium G30. Arch Microbiol 109: 51–58 ( 1976)

    CAS  Article  Google Scholar 

  6. 6

    Moneib NA . In-vitro activity of commonly used antifungal agents in the presence of rifampin, polymyxin B and norfloxacin against Candida albicans. J Chemother 7: 525–529 ( 1995)

    CAS  Article  Google Scholar 

  7. 7

    Schwartz SN, Medoff G, Kobayashi GS, Kwan CN, Schlessinger D . Antifungal properties of polymyxin B and its potentiation of tetracycline as an antifungal agent. Antimicrob Agents Chemother 2: 36–40 ( 1972)

    CAS  Article  Google Scholar 

  8. 8

    Boguslawski G . Effects of polymyxin B sulfate and polymyxin B nonapeptide on growth and permeability of the yeast Saccharomyces cerevisiae. Mol Gen Genet 199: 401–405 ( 1985)

    CAS  Article  Google Scholar 

  9. 9

    Nicholls MWN . Polymyxin sensitivity of Candida tropicalis. J Med Microbiol 3: 529–538 ( 1970)

    CAS  Article  Google Scholar 

  10. 10

    Miron T, Rabinkov A, Mirelman D, Wilchek M, Weiner L . The mode of action of allicin: its ready permeability through phospholipid membranes may contribute to its biological activity. Biochim Biophys Acta 1463: 20–30 ( 2000)

    CAS  Article  Google Scholar 

  11. 11

    Agarwal KC . Therapeutic actions of garlic constituents. Med Res Rev 16: 111–124 ( 1996)

    CAS  Article  Google Scholar 

  12. 12

    Ankri S, Mirelman D . Antimicrobial properties of allicin from garlic. Microbes Infect 1: 125–129 ( 1999)

    CAS  Article  Google Scholar 

  13. 13

    Chu TC, Ogidigben M, Han JC, Potter DE . Allicin-induced hypotension in rabbit eyes. J Ocul Pharmacol 9: 201–209 ( 1993)

    CAS  Article  Google Scholar 

  14. 14

    Ha MW, Yuan Y . Allicin induced cell cycle arrest in human gastric cancer cell lines. Zhonghua Zhong Liu Za Zhi 26: 585–589 ( 2004)

    CAS  PubMed  Google Scholar 

  15. 15

    Oommen S, Anto RJ, Srinivas G, Karunagaran D . Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells. Eur J Pharmacol 485: 97–103 ( 2004)

    CAS  Article  Google Scholar 

  16. 16

    Rabinkov A, Miron T, Mirelman D, Wilchek M, Glozman S, Yavin E, Weiner L . S-Allylmercaptoglutathione: the reaction product of allicin with glutathione possesses SH-modifying and antioxidant properties. Biochim Biophys Acta 1499: 144–153 ( 2000)

    CAS  Article  Google Scholar 

  17. 17

    Ankri S, Miron T, Rabinkov A, Wilchek M, Mirelman D . Allicin from garlic strongly inhibits cysteine proteinases and cytopathic effects of Entamoeba histolytica. Antimicrob Agents Chemother 41: 2286–2288 ( 1997)

    CAS  Article  Google Scholar 

  18. 18

    Ogita A, Fujita K, Taniguchi M, Tanaka T . Enhancement of the fungicidal activity of amphotericin B by allicin, an allyl-sulfur compound from garlic, against the yeast Saccharomyces cerevisiae as a model system. Planta Med 72: 1247–1250 ( 2006)

    CAS  Article  Google Scholar 

  19. 19

    Davis LE, Shen J, Royer RE . In vitro synergism of concentrated Allium sativum extract and amphotericin B against Cryptococcus neoformans. Planta Med 60: 546–549 ( 1994)

    CAS  Article  Google Scholar 

  20. 20

    Tanaka T, Nakayama K, Machida K, Taniguchi M . Long-chain alkyl ester of AMP acts as an antagonist of glucose-induced signal transduction that mediates activation of plasma membrane proton pump in Saccharomyces cerevisiae. Microbiology 146: 377–384 ( 2000)

    CAS  Article  Google Scholar 

  21. 21

    Ramotowski S, Szczesniak M . Determination of potassium salt content in pharmaceutical preparations by means of sodium tetraphenylborate. Acta Pol Pharm 24: 605–613 ( 1967)

    CAS  PubMed  Google Scholar 

  22. 22

    Vida TA, Emr SD . A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol 128: 779–792 ( 1995)

    CAS  Article  Google Scholar 

  23. 23

    Danner RL, Joiner KA, Rubin M, Patterson WH, Johnson N, Ayers KM, Parrillo JE . Purification, toxicity, and antiendotoxin activity of polymyxin B nonapeptide. Antimicrob Agents Chemother 33: 1428–1434 ( 1989)

    CAS  Article  Google Scholar 

  24. 24

    Galizzi A, Cacco G, Siccardi AG . Mazza G. Mode of action of polymyxin: Physiological studies with Bacillus subtilis-resistant mutant. Antimicrob Agents Chemother 8: 366–369 ( 1975)

    CAS  Article  Google Scholar 

  25. 25

    Amagase H, Petesch BL, Matsuura H, Kasuga S, Itakura Y . Intake of garlic and its bioactive components. J Nutr 131: 955–962 ( 2001)

    Article  Google Scholar 

  26. 26

    Daugelavicius R, Bakiene E, Bamford DH . Stages of polymyxin B interaction with the Escherichia coli cell envelope. Antimicrob Agents Chemother 44: 2969–2678 ( 2000)

    CAS  Article  Google Scholar 

  27. 27

    Avery SV, Howlett NG, Radice S . Copper toxicity towards Saccharomyces cerevisiae: dependence on plasma membrane fatty acid composition. Appl Environ Microbiol 62: 3960–3966 ( 1996)

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28

    Nakayama K, Yamaguchi T, Doi T, Usuki Y, Taniguchi M, Tanaka T . Synergistic combination of direct plasma membrane damage and oxidative stress as a cause of antifungal activity of polyol macrolide antibiotic niphimycin. J Biosci Bioeng 94: 207–211 ( 2002)

    CAS  Article  Google Scholar 

  29. 29

    Obara K, Kuriyama H, Fukuda H . Direct evidence of active and rapid nuclear degradation triggered by vacuole rupture during programmed cell death in Zinnia. Plant Physiol 125: 615–626 ( 2001)

    CAS  Article  Google Scholar 

  30. 30

    Ogita A, Hirooka K, Yamamoto Y, Tsutsui N, Fujita K, Taniguchi M, Tanaka T . Synergistic fungicidal activity of Cu2+ and allicin, an allyl sulfur compound from garlic, and its relation to the role of alkyl hydroperoxide reductase 1 as a cell surface defense in Saccharomyces cerevisiae. Toxicology 215: 205–213 ( 2005)

    CAS  Article  Google Scholar 

  31. 31

    Ogita A, Fujita K, Taniguchi M, Tanaka T . Dependence of synergistic fungicidal activity of Cu2+ and allicin, an allyl sulfur compound from garlic, on selective accumulation of the ion in the plasma membrane fraction via allicin-mediated phospholipid peroxidation. Planta Med 72: 875–880 ( 2006)

    CAS  Article  Google Scholar 

  32. 32

    Ogita A, Matsumoto K, Fujita K, Usuki Y, Hatanaka Y, Tanaka T . Synergistic fungicidal activities of amphotericin B and N-methyl-N″-dodecylguanidine: a constituent of polyol macrolide antibiotic niphimycin. J Antibiot 60: 27–35 ( 2007)

    CAS  Article  Google Scholar 

  33. 33

    Arnold TM, Forrest GN, Messmer KJ . Polymyxin antibiotics for Gram-negative infections. Am J Health Syst Pharm 64: 819–826 ( 2007)

    CAS  Article  Google Scholar 

  34. 34

    Carrillo-Muñoz AJ, Giusiano G, Ezkurra PA, Quindós G . Antifungal agents: Mode of action in yeast cells. Rev Esp Quimioter 19: 130–139 ( 2006)

    PubMed  Google Scholar 

  35. 35

    Banta LM, Robinson JS, Klionsky DJ, Emr SD . Organelle assembly in yeast: Characterization of yeast mutants defective in vacuolar biogenesis and protein sorting. J Cell Biol 107: 1369–1383 ( 1988)

    CAS  Article  Google Scholar 

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Correspondence to Toshio Tanaka.

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Ogita, A., Nagao, Y., Fujita, Ki. et al. Amplification of Vacuole-targeting Fungicidal Activity of Antibacterial Antibiotic Polymyxin B by Allicin, an Allyl Sulfur Compound from Garlic. J Antibiot 60, 511–518 (2007).

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  • polymyxin B
  • allicin
  • Saccharomyces cerevisiae
  • vacuole
  • plasma membrane

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