Characteristics of Sophorolipid as an Antimicrobial Agent

  • KIM, KAPJUNG (Department of Biological Engineering, Inha University) ;
  • DALSOO YOO (Department of Biological Engineering, Inha University) ;
  • YOUNGBUM KIM (Department of Biological Engineering, Inha University) ;
  • BAEKSEOK LEE (Department of Biological Engineering, Inha University) ;
  • DOONHOON SHIN (Graduate School of Biotechnology, Korea University) ;
  • EUN-KI KIM (Department of Biological Engineering, Inha University)
  • Published : 2002.04.01


Sophorolipid, a biosurfactant produced from Candida bombicola ATCC 22214, showed antimicrobial activity against Bacillus subtilis, Staphylococcus xylosus, Streptococcus mutans, and Propionibacterium acne at 4, 1, 1, 0.5 ppm, respectively. Also, 100 ppm of sophorolipid inhibited $50\%$ of cell growth of plant pathogenic fungus, Botrytis cineria. However, sophorolipid showed no effect on Escherichia coli, indicating that its selective antimicrobial activity depended on the cell wall structure. Treatment of B. subtilis with sophorolipid increased leakage of intracellular enzyme, malate dehydrogenase, indicating a possible interaction of sophorolipid with a cellular membrane. Comparing lactone-type and acid-type sophorolipids, the former showed a higher antimicrobial activity. Supplementing other surfactants showed no significant effects on the antimicrobial activity. Animal study showed that 5 g of sophorolipid per kg body weight by oral administration caused no toxicity, and sophorolipid induced no irritation on the skin. These results show potential use of sophorolipid as an active ingredient in healthcare products.


  1. Arima, K., A. Kakinuma, and G. Tamura. 1968. Surfactin, a crystaline peptide-lipid surfactant produced by Bacillus subtilis: Isolation, characterization and its inhibition of fibrin clot formation. Biochem. Biophys. Res. Commun. 31: 488- 494
  2. Asmer, H.-J., S. Lang, F. Wagner, and V. Wray. 1988. Microbial production, structure and elucidation and bioconversion of sophorose lipids. J. Am. Oil Chem. Soc. 65: 1460-1466
  3. Berbheimer, A. W. and L. S. Avigad. 1970. Nature and properties of a cytological agent produced by Bacillus subtilis. J. Gen. Microbiol. 61: 361-369
  4. Bezkorovainy, A. 1981. Antimicrobial properties of ironbinding proteins. Adv. Exp. Med. Biol. 135: 139-154
  5. Bhushan, B. and G. S. Hoondal. 1999. Effect of fungicides, insecticides and allosamidin on a thermostable chitinase from Bacillus sp. BG-11. World J. Microbiol. Biotechnol. 15: 403-404
  6. Blanc, P. and G. Goma. 1987. Kinetics of inhibition in propionic acid fermentation. Bioprocess Eng. 2: 175-179
  7. Brakemeier, A., D. Wullbrandt, and S. Lang. 1998. Candida bombicola: Production of novel alkyl-glycosides based on glucose/2-dodecanol. Appl. Microbiol. Biotechnol. 50: 161-166
  8. Brakemeier, A., D. Wullbrandt, and S. Lang. 1998. Microbial alkyl-sophorosides based on 1-dodecanol or 2-, 3-, and 4- dodecanones. Biotechnol. Lett. 20: 215-218
  9. Brakemeier, A., S. Lang, D. Wullbrandt, L. Merschel, A. Benninghoven, N. Buschmann, and F. Wagner. 1995. Novel sophorose lipids from microbial conversion of 2-alkanols. Biotechnol. Lett. 17: 1183-1188
  10. Ching. J. U., W. Sheu, and E. Freese. 1972. Effects of fatty acids on growth and envelope proteins of Bacillus subtilis. J. Bacteriol. 111: 516-524
  11. Cho, K.-J., Y. B. Kim, and E. K. Kim. 1999. Production and application of sophorolipid, a microbial surfactant. Kor. J. Biotechnol. Bioeng. 14: 747-753
  12. Choi, K.-S., S. H. Kim, and T. H. Lee. 1999. Purification and characterization of biosurfactant from Tsukamurella sp. 26A. J. Microbiol. Biotechnol. 9: 32-38
  13. Cooper, A. J. 1998. Systematic review of Propionibacterium acne resistance to systemic antibiotics. Med. J. Australia 169: 259-261
  14. Daniel, H. J. and R. T. Otto. 1999. Production of sophorolipids from whey: Development of a two-stage process with Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214 using deproteinized whey concentrates as substrates. Appl. Microbiol. Technol. 25: 613-621
  15. Davila, A. M., R. Marchal, and J. P. Vandecasteele. 1997. Sophorose lipid fermentation with differentiated substrate supply for growth and production phases. Appl. Microbiol. Biotechnol. 47: 496-501
  16. Desai, J. P. and I. M. Banat. 1997. Microbial production of surfactants and their commercial potential. Microbiol. Mol. Rev. 61: 47-64
  17. Finnerty, W. R. 1994. Biosurfactant in environmental biotechnology. Curr. Opin. Biotechnol. 5: 291-295
  18. Food and Drug Association, 1999. Standard Methods for Toxicity Test of Pharmaceuticals; 1999-61, Korea
  19. Gang, J.-G., H. C. Yun, J. C. Son, and S. Y. Hwong. 1998. Effect of 1-[(2-hydroxyethoxy) methyl]-6-(phenylthio) thymine (HEPT) derivatives on bacterial growth. J. Microbiol. Biotechnol. 8: 229-236.
  20. Gribbon, E. M., W. J. Cunliffe, and K. T. Holland. 1993. Interaction of Propionibacterium acnes with skin lipids in vitro. J. Gen. Microbiol. 139: 1745-1751
  21. Han, C.-S., H. S. Yun, H. J. Seo, and E. K. Kim. 1999. Biosurfactant production from phenanthrene degrading bacteria. Kor. J. Biotechnol. Bioeng. 14: 737-741
  22. Hommel, R. K., O. Stuwer, W. Stuber, D. Haferberg, and H. P. Kleber. 1987. Production of water-soluble surface active exolipids by Torulopsis apicola. Appl. Microbiol. Biotechnol. 26: 199-205
  23. Ito, S. and S. Inoue. 1982. Sophorolipids from Torulopsis bombicola: Possible relation to alkane uptake. Appl. Environ. Microbiol. 43: 1278-1283
  24. Itokawa, H., T. Miyashita, H. Morita, K. Takeya, T. Hirano, M. Homma, and O. Kitaro. 1994. Structural and conformational studies of Ile7 and Leu7 surfactant from Bacillus subtilis. Chem. Pharm. Bull. 42: 604-607
  25. Jung, H.-K., J. B. Lee, G. B. Yim, and E. K. Kim. 1995. Properties of microbial surfactant, S-acid. Kor. J. Biotechnol. Bioeng. 10: 71-77
  26. Jung, H.-K., J. B. Lee, G. B. Yim, and E. K. Kim. 1995. Properties of microbial surfactant, S-acid. Kor. J. Biotechnol. Bioeng. 10: 71-77
  27. Kim, D.-H., J. B. Lee, G. B. Yim, and E. K. Kim. 1995. Optimized production of microbial surfactant, S-acid, from Penicillium spiculisporum. Kor. J. Biotechnol. Bioeng. 10: 113-118
  28. Kim, J.-S., M. Powalla, S. Lang, F. Wagner, H. Lünsdorf, and V. Wray. 1990. Microbial glycolipid production under nitrogen limitation and resting cell conditions. J. Biotechnol. 13: 257-266
  29. Kim, S.-H., J. D. Lee, B. C. Lee, B. C. Kim, and T. H. Lee. 1996. Purification and characterization of bioemulsifier produced by Acinetobacter sp. BE-254. J. Microbiol. Biotechnol. 6: 184-188
  30. Koh, K. H. 1995. Antifungal activity of caproic acid against Botrytis cinerea. Kor. J. Appl. Microbiol. Biotechnol. 23: 378-383
  31. Kosaric, N., W. L. Cairns, N. C. C. Gray, D. Stechey, and J. Wood. 1984. The role of nitrogen in multiorganism strategies for biosurfactant production. J. Am. Oil Chem. Soc. 61: 1735-1743
  32. Lahov, E. and W. Regelson. 1996. Antibacterial and immunostimulating casein-derived substances from milk: Casecidin, isracidin peptides. Food Chem. Toxicol. 34: 131- 145.
  33. Lang, S., A. Brakemeier, A. Schlotterbeck, and F. Wagner. 1996. Biotechnological synthesis of surface-active glycolipids. Euro Cosmetics 4: 41-45
  34. Lang, S., E. Katsiwela, and F. Wagner. 1989. Antimicrobial effects of biosurfactants. Fat Sci. Technol. 91: 363-366
  35. Lee, B.-Y., I. H. Yoon, Y. B. Kim, P. J. Han, and C. M. Lee. 1985. Studies on storing chest-nut (Castanea crenata var. dulcis Nakai) sealing with polyethylene film. Korean J. Food Sci. Technol. 17: 331-335
  36. Lee, H.-Y., C. K. Kim, T. K. Sung, T. K. Mun, and C. J. Lim. 1992. Antibacterial activity of Ulmus pumila L. extract. Kor. J. Appl. Microbiol. Biotechnol. 20: 1-5
  37. Lin, S. C. 1996. Biosurfactants: Recent advances. J. Chem. Tech. Biotechnol. 66: 109-120<109::AID-JCTB477>3.0.CO;2-2
  38. Miller, S. S., B. T. Driscoll, R. G. Gregerson, J. S. Gantt, and C. P. Vance. 1998. Alfalfa malate dehydrogenase MDH molecular cloning and characterization of five different forms reveals a unique nodule-enhanced MDH. Plant J. 15: 173-184
  39. Mitsubishi-Kasei Food Corporation, 1987. Ryoto Sugar Ester, Technical Information, Mitsubishi-Kasei Food Corporation, Japan
  40. Muller-Hurtig, R., F. Wagner, R. Blaszczyk, and N. Kosaric. 1993. Biosurfactants for environmental control, pp. 447- 469. In N. Kosaric (ed.), Biosurfactant - Production, Properties and Applications. Surfactant Science Series, Vol. 48. Marcel Dekker, New York, U.S.A
  41. Naruse, N., O. Tenmyo, S. Kobaru, H. Kamai, T. Miyaki, M. Konishi, and T. Oki. 1990. Pumilacidin, a complex of new antiviral antibiotics: Production, isolation, chemical properties, structure and biological activity. J. Antibiot. 43: 267-280
  42. Nikolaos, E. L. and D. C. Yannis. 1997. L-Malate dehydrogenase from Pseudomonas stuzeri: Purification and characterization. Archives Biochem. Biophys. 337: 103-114
  43. Ocheretina, O. and R. Scheibe. 1997. Cloning and sequence analysis of cDNAs encoding plant cytosolic malate dehydrogenase. Gene 199: 145-148
  44. Rau, U., R. Heckmann, V. Wray, and S. Lang. 1999. Enzymatic conversion of a sophorose lipid into a glucose lipid. Biotechnol. Lett. 21: 973-977
  45. Rau, U., S. Hammen, R. Heckmann, V. Wray, and S. Lang. 2001. Sophorolipids: A source for novel compounds. Ind. Crops Products 13: 85-92
  46. Stanghellini, M. E. and R. E. Miller. 1999. Biosurfactants: Their identity and potential efficacy in the biological control of zoosporic plant pathogens. Plant Dis. 81: 4-12
  47. Suk, W.-S., H. J. Son, G. Lee, and S. J. Lee. 1999. Purification and characterization of biosurfactants produced by Pseudomonas sp. SW1. J. Microbiol. Biotechnol. 9: 56- 61
  48. Sullivan, E. R. 1998. Molecular genetics of biosurfactant production. Curr. Opin. Biotechnol. 9: 263-269
  49. Tagesson, C. and C. Edling. 1984. Influence of surfaceactive food additives on the integrity and permeability of rat intestinal mucosa. Food Chem. Toxicol. 22: 861-864
  50. Tulloch, A. P., A. Hill, and J. F. T. Spencer. 1968. Structure and reactions of lactonic and acidic sophorosides of 17- hydroxy octadecanoic acid. Can. J. Chem. 46: 3337-3351
  51. Tulloch, A. P. 1976. Structures of extracellular glycolipids produced by yeasts, pp. 329-345. In L. A. Witting (ed.), Glycolipid Methodology. American Oil Chemists Society, Champaign, IL, U.S.A
  52. Velikonja, J. and N. Kosaric. 1993. Biosurfactants in food application, pp. 419-446. In N. Kosaric (ed.), Biosurfactant - Production, Properties and Applications. Marcel Dekker, New York, U.S.A
  53. Watanabe T., S. Katayama, M. Matsubara, Y. Honda, and M. Kuwahara. 2000. Antibacterial carbohydrate monoesters suppressing cell growth of Streptococcus mutans in the presence of sucrose. Curr. Microbiol. 41: 210-213
  54. Zheng, J. and V. D. Ramirez. 2000. Inhibition of mitochondrial proton F0F1-ATPase/ATP synthase by polyphenolic phytochemicals. Br. J. Pharmacol. 130: 1115-1123