Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Isolation, Characterization, and Investigation of Surface and Hemolytic Activities of a Lipopeptide Biosurfactant Produced by Bacillus subtilis ATCC 6633

  • Published : 2005.06.01
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Abstract

Bacillus subtilis ATCC 6633 was grown in BHIB medium supplemented with $Mn^{2+}$ for 96 h at $37^{\circ}C$ in a shaker incubator. After removing the microbial biomass, a lipopeptide biosurfactant was extracted from the supernatant. Its structure was established by chemical and spectroscopy methods. The structure was confirmed by physical properties, such as Hydrophile-Lipophile Balance (HLB), surface activity and erythrocyte hemolytic capacity. The critical micelle concentration (cmc) and erythrocyte hemolytic capacity of the biosurfactant were compared to those of surfactants such as SDS, BC (benzalkonium chloride), TTAB (tetradecyltrimethylammonium bromide) and HTAB (hexadecyltrimethylammonium bromide). The maximum hemolytic effect for all surfactants mentioned was observed at concentrations above cmc. The maximum hemolytic effect of synthetic surfactants was more than that of the biosurfactant produced by B. subtilis ATCC 6633. Therefore, biosurfactant would be considered a suitable surface-active agent due to low toxicity to the membrane.

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References

  1. Arima, K., A. Kakinuma, and G. Tamura. 1968. Surfactin, a crystalline peptidelipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochem. Biophys. Res. Commun. 31, 488-494 https://doi.org/10.1016/0006-291X(68)90503-2
  2. Banat, I.M., R.S. Makkar, and S.S. Cameotra. 2000. Potential applications of microbial surfactants. Appl. Microbiol. Biotechnol. 53, 495-508 https://doi.org/10.1007/s002530051648
  3. Carrillo, P.G., C. Mardaraz, S.I. Pitta-Alvarez, and A.M. Giulietti. 1996. Isolation and selection of biosurfactant producing bacteria. World J. Microbiol. Biotechnol. 12, 82-84 https://doi.org/10.1007/BF00327807
  4. Cooper, D.G., C.R. Macdonald, S.J. Duff, and N. Kosaric. 1981. Enhanced production of surfactin from Bacillus subtilis by continuous product removal and metal cation additions. Appl. Environ. Microbiol. 42, 408-412
  5. Desai, J.D. and I.M. Banat. 1997. Microbial production of surfactant and their commercial potential. Microbiol. Mol. Biol. 61, 47-64
  6. Feignier, C., F. Besson, and G. Michel. 1995. Studies on lipopeptide biosynthesis by Bacillus subtilis: isolation and characterization of iturin-, surfactin+ mutants. FEMS Microbiol. Lett. 127, 11-15
  7. Gould, L.A., A.B. Lansley, M.B. Brown, B. Forbes, and G.P. Martin. 2000. Mitigation of surfactant erythrocyte toxicity by egg phophatidylcholine. J. Pharm. Pharmacol. 52, 1203-1209 https://doi.org/10.1211/0022357001777333
  8. Heerklotz, H. and J. Seelig. 2001. Detergent-like action of the antibiotic peptide surfactin on lipid membranes. Biophys. J. 81, 1547-1554 https://doi.org/10.1016/S0006-3495(01)75808-0
  9. Ishigami, Y. 1997. Characterization of biosurfactants, p. 197-226. In K. Esmi (ed.), Structure-performance relationships in surfactants. Dekker Press, New York
  10. Kayes, J.B. 1989. Disperse systems, p. 99-117. In M.E. Aulton (ed.), Pharmaceutics, The science of dosage form design, Churchill livingstone, London
  11. Makkar, R.S. and S.S. Cameotra. 1999a. Biosurfactant production by microorganisms on unconventional carbon sources - a review. J. Surf. Det. 2, 237-241 https://doi.org/10.1007/s11743-999-0078-3
  12. Nissen, E., J. Vater, G. Pauli, and D. Vollenbroich. 1997. Application of surfactin for mycoplasma inactivation in virus stocks. In Vitro Cell Dev. Biol. Anim. 33, 414-41 https://doi.org/10.1007/s11626-997-0056-8
  13. Orafidiya, L.O. and F.A. Oladimeji. 2002. Determination of the required HLB values of some essential oils. Int. J. Pharm. 237, 241-249 https://doi.org/10.1016/S0378-5173(02)00051-0
  14. Peypoux, F., J.M. Bonmatin, and J. Wallach. 1999. Recent trends in the biochemistry of surfactin. Appl. Microbiol. Biotechnol. 51, 553-563 https://doi.org/10.1007/s002530051432
  15. Peypoux, F., J.M. Bonmatin, H. Labbe, I. Grangemard, B.C. Das, M. Tak, J. Wallach, and G. Michel. 1994. [Ala 4] surfactin, a novel isoform from Bacillus subtilis studied by mass and NMR spectroscopies. Eur. J. Biochem. 224, 89-96. https://doi.org/10.1111/j.1432-1033.1994.tb19998.x
  16. Provencher, S. and J. Glockner. 1981. Estimation of globular protein secondary structure from Circular dichroism. Biochem. J. 20, 33-37 https://doi.org/10.1021/bi00504a006
  17. Randhir, S.M. 1997. Utilization of molasses for biosurfactant production by two Bacillus strains at thermophilic conditions. JAOCS 74, 887-889 https://doi.org/10.1007/s11746-997-0233-7
  18. Razafindralambo, H., Y. Popineau, M. Deleu, C. Hbid, P. Jacques, P. Thonart, and M. Paquot. 1998. Foaming properties of lipopeptides produced by Bacillus subtilis: effect of lipid and peptide structural attributes. J. Agric. Food Chem. 46, 911-916 https://doi.org/10.1021/jf970592d