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Current IssueEditorial BoardOnline First ArticlesArchive

volume 31 issue 4 (december 2011) : 239 - 246

ANTIMICROBIAL ACTIVITY OF PURIFIED EMULSIFIER OF ACINETOBACTER GENOSPECIES ISOLATED FROM RHIZOSPHERE OF WHEAT

S
S.D. Bhawsar*
S
S.D. Patil1
B
B.A. Chopade2
1Department of Microbiology, University of Pune, Pune Maharashtra – 411 001, India.

  • Submitted|

  • First Online |

  • doi

Cite article:- Bhawsar* S.D., Patil1 S.D., Chopade2 B.A. (2025). ANTIMICROBIAL ACTIVITY OF PURIFIED EMULSIFIER OF ACINETOBACTER GENOSPECIES ISOLATED FROM RHIZOSPHERE OF WHEAT. Agricultural Science Digest. 31(4): 239 - 246. doi: .

ABSTRACT

Among thirty seven wheat rhizosphere isolates, only Acinetobacter genospecies 3, A15 emulsified all thirty oil substrates and combinations in LB medium. It emulsified diesel oil wide range of temperatures 00C - 400C, pH 1.5-12, 5-25% salinity with EI30Days of 75.9% at 00C. Emulsification activity was comparable to SDS, Tween 20 and 80, Triton X100 surfactants and was not affected by chelators and enzymes. Partially purified bioemulsifier was composed of 50% polysaccharide, 30% protein and 2% lipid. It was not surface active but showed activity against human pathogens, Staphylococcus aureus, Salmonella typhimurium, Klebsiella pneumoniae, Aspergillus niger, Aspergillus fumigatus, Candida humicola, Candida albicans, Cryptococcus and inhibited PS and Wish viral cell lines. Purified compound at R.T.1.440 with diesel emulsification activity was obtained by HPLC. Infrared spectrum confirmed OH, N-H or COOH; C=O; C=C or Ar or C-H; NO2 or Ar-O; Ar-C-O; C-O functional groups. GC-MS and 1HNMR analysis respectively indicated methyl esters and methylated fatty acids as structural components of bioemulsifier. Microbicidal activity of purified bioemulsifier against C. albicans was observed microscopically.

REFERENCES

  1. Amenta, J. S. (1964). A rapid chemical method for quantification of lipids separated by thin layer chromatography. J. Lipid Res. 5: 270-272.
  2. Arima K., Kakinuma, A. and Tamura, G. C. (1968). Surfactin, a crystalline peptide lipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochem. Biophys. Res. Commun. 31: 488-494.
  3. Cooper, D. G. and Goldenberg, B.G. (1987). Surface active agents from two Bacillus sp. Appl. Environ. Microbiol. 53: 224-229.
  4. Desai, J. D. (1987). Microbial surfactants: evaluation, types, production and future applications. J. Sci. Ind. Res. 46: 440-449.
  5. Desai, A. J., Patel, R. M. and Desai, J. D. (1994). Advances in production of biosurfactants and their commercial applications. J. Sci. Ind. Res. 53: 619-629.
  6. Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. and Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Anal. Biochem. 28 (3) : 350-356.
  7. Huddedar, S. B., A. M., Tilekar, J. N., Gore, S. D., Dhavale, D. D. and Chopade, B. A. (2002). Isolation, characterization and plasmid pUPI126 mediated indole-3-acetic acid production in Acinetobacter strains from rhizosphere of wheat. Appl. Biochem. Biotechnol. 102-103 : 21-39.
  8. Kaplan, N. and Rosenberg, E. (1982). Exopolysaccharide distribution of and bioemulsifier production by Acinetobacter calcoaceticus BD4 and BD413. Appl. Environ. Microbiol. 44 (6) : 1335-1341.
  9. Kim, S. H., Lim, E. J., Lee, S. O., Lee, J. D. and Lee, T. H. (2000). Purification and characterization of biosurfactants from Nocardia sp. L-417. Biotechnol. Appl. Biochem. 31 : 249-253.
  10. Lowry, O. H., Rosenbrough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with Folin phenol reagent. J. Biol. Chem. 193 : 256-275.
  11. Morikawa, M., Daido, H., Takao, T., Murata, S., Shimonishi, Y. and Imanaka, T. (1993). A new lipopeptide biosurfactant produced by Arthrobacter sp. strain MIS38. J. Bacteriol. 175 (20) : 6459-6466.
  12. Neufeld, R. J. and Zajic, J. E. (1984). Surface activity of Acinetobacter calcoaceticus sp. 2CA2. Biotechnol. Bioeng. 26 : 1108-1113.
  13. Patil, J. R. and Chopade, B. A. (2001). Studies on bioemulsifier production by Acinetobacter strains isolated from healthy human skin. J. Appl. Microbiol. 91: 290-298.
  14. Pines, O., Bayer, E. A., Gutnick, D. L. (1983). Localization of emulsan like polymers associated with the cell surface of Acinetobacter calcoaceticus. J.Bacteriol. 154 (2) : 893-905.
  15. Rosenberg, E. (1986). Microbial surfactants. Crit. Rev. Biotechnol. 3: 109-132.
  16. Rosenberg, E. and Ron, E. Z. (1997). Bioemulsions: microbial polymeric emulsifiers. Curr. Opin. Biotechnol. 8: 313-316.
  17. Singh, P. and Cameotra, S. S. (2004). Potential applications of microbial surfactants in biomedical sciences. Trends Biotechnol. 22 (3) : 142-146.
  18. Venezia, S. N., Zosim, Z., Gottlieb, A., Legmann, R., Carmeli, S., Ron, E. Z. and Rosenberg, E. (1995). Alasan, a new bioemulsifier from Acinetobacter radioresistens. Appl. Environ. Microbiol. 61 (9) : 3240-3244.
  19. Zuckerberg, A., Peeri, D. Z., Gutnick, D. L. and Rosenberg, E. (1979). Emulsifier of Arthrobacter RAG –1: chemical and physical properties. Appl. Environ. Microbiol. 37 (3) : 414-420.
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    volume 31 issue 4 (december 2011) : 239 - 246

    ANTIMICROBIAL ACTIVITY OF PURIFIED EMULSIFIER OF ACINETOBACTER GENOSPECIES ISOLATED FROM RHIZOSPHERE OF WHEAT

    S
    S.D. Bhawsar*
    S
    S.D. Patil1
    B
    B.A. Chopade2
    1Department of Microbiology, University of Pune, Pune Maharashtra – 411 001, India.

    • Submitted|

    • First Online |

    • doi

    Cite article:- Bhawsar* S.D., Patil1 S.D., Chopade2 B.A. (2025). ANTIMICROBIAL ACTIVITY OF PURIFIED EMULSIFIER OF ACINETOBACTER GENOSPECIES ISOLATED FROM RHIZOSPHERE OF WHEAT. Agricultural Science Digest. 31(4): 239 - 246. doi: .

    ABSTRACT

    Among thirty seven wheat rhizosphere isolates, only Acinetobacter genospecies 3, A15 emulsified all thirty oil substrates and combinations in LB medium. It emulsified diesel oil wide range of temperatures 00C - 400C, pH 1.5-12, 5-25% salinity with EI30Days of 75.9% at 00C. Emulsification activity was comparable to SDS, Tween 20 and 80, Triton X100 surfactants and was not affected by chelators and enzymes. Partially purified bioemulsifier was composed of 50% polysaccharide, 30% protein and 2% lipid. It was not surface active but showed activity against human pathogens, Staphylococcus aureus, Salmonella typhimurium, Klebsiella pneumoniae, Aspergillus niger, Aspergillus fumigatus, Candida humicola, Candida albicans, Cryptococcus and inhibited PS and Wish viral cell lines. Purified compound at R.T.1.440 with diesel emulsification activity was obtained by HPLC. Infrared spectrum confirmed OH, N-H or COOH; C=O; C=C or Ar or C-H; NO2 or Ar-O; Ar-C-O; C-O functional groups. GC-MS and 1HNMR analysis respectively indicated methyl esters and methylated fatty acids as structural components of bioemulsifier. Microbicidal activity of purified bioemulsifier against C. albicans was observed microscopically.

    REFERENCES

    1. Amenta, J. S. (1964). A rapid chemical method for quantification of lipids separated by thin layer chromatography. J. Lipid Res. 5: 270-272.
    2. Arima K., Kakinuma, A. and Tamura, G. C. (1968). Surfactin, a crystalline peptide lipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochem. Biophys. Res. Commun. 31: 488-494.
    3. Cooper, D. G. and Goldenberg, B.G. (1987). Surface active agents from two Bacillus sp. Appl. Environ. Microbiol. 53: 224-229.
    4. Desai, J. D. (1987). Microbial surfactants: evaluation, types, production and future applications. J. Sci. Ind. Res. 46: 440-449.
    5. Desai, A. J., Patel, R. M. and Desai, J. D. (1994). Advances in production of biosurfactants and their commercial applications. J. Sci. Ind. Res. 53: 619-629.
    6. Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. and Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Anal. Biochem. 28 (3) : 350-356.
    7. Huddedar, S. B., A. M., Tilekar, J. N., Gore, S. D., Dhavale, D. D. and Chopade, B. A. (2002). Isolation, characterization and plasmid pUPI126 mediated indole-3-acetic acid production in Acinetobacter strains from rhizosphere of wheat. Appl. Biochem. Biotechnol. 102-103 : 21-39.
    8. Kaplan, N. and Rosenberg, E. (1982). Exopolysaccharide distribution of and bioemulsifier production by Acinetobacter calcoaceticus BD4 and BD413. Appl. Environ. Microbiol. 44 (6) : 1335-1341.
    9. Kim, S. H., Lim, E. J., Lee, S. O., Lee, J. D. and Lee, T. H. (2000). Purification and characterization of biosurfactants from Nocardia sp. L-417. Biotechnol. Appl. Biochem. 31 : 249-253.
    10. Lowry, O. H., Rosenbrough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with Folin phenol reagent. J. Biol. Chem. 193 : 256-275.
    11. Morikawa, M., Daido, H., Takao, T., Murata, S., Shimonishi, Y. and Imanaka, T. (1993). A new lipopeptide biosurfactant produced by Arthrobacter sp. strain MIS38. J. Bacteriol. 175 (20) : 6459-6466.
    12. Neufeld, R. J. and Zajic, J. E. (1984). Surface activity of Acinetobacter calcoaceticus sp. 2CA2. Biotechnol. Bioeng. 26 : 1108-1113.
    13. Patil, J. R. and Chopade, B. A. (2001). Studies on bioemulsifier production by Acinetobacter strains isolated from healthy human skin. J. Appl. Microbiol. 91: 290-298.
    14. Pines, O., Bayer, E. A., Gutnick, D. L. (1983). Localization of emulsan like polymers associated with the cell surface of Acinetobacter calcoaceticus. J.Bacteriol. 154 (2) : 893-905.
    15. Rosenberg, E. (1986). Microbial surfactants. Crit. Rev. Biotechnol. 3: 109-132.
    16. Rosenberg, E. and Ron, E. Z. (1997). Bioemulsions: microbial polymeric emulsifiers. Curr. Opin. Biotechnol. 8: 313-316.
    17. Singh, P. and Cameotra, S. S. (2004). Potential applications of microbial surfactants in biomedical sciences. Trends Biotechnol. 22 (3) : 142-146.
    18. Venezia, S. N., Zosim, Z., Gottlieb, A., Legmann, R., Carmeli, S., Ron, E. Z. and Rosenberg, E. (1995). Alasan, a new bioemulsifier from Acinetobacter radioresistens. Appl. Environ. Microbiol. 61 (9) : 3240-3244.
    19. Zuckerberg, A., Peeri, D. Z., Gutnick, D. L. and Rosenberg, E. (1979). Emulsifier of Arthrobacter RAG –1: chemical and physical properties. Appl. Environ. Microbiol. 37 (3) : 414-420.
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