Skip to main content
Log in

Emulsifier production and microscopical study of emulsions and biofilms formed by the hydrocarbon-utilizing bacteria Acinetobacter calcoaceticus MM5

  • Environmental biotechnology
  • Original Paper
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A bacterial strain was isolated from a sample of contaminated heating oil and identified as a strain of Acinetobacter calcoaceticus, named MM5. The bacterial isolate was able to grow on petroleum derivatives and brought about an emulsification of those compounds. A bioemulsifier was extracted from the culture medium of MM5 strain and partially characterized. This compound was able to emulsify petroleum fuels and both aliphatic and aromatic pure hydrocarbons and was stable over a wide range of temperatures. Studies developed by light, scanning electron and transmission electron microscopy showed that, during the growth on petroleum derivatives, the microorganisms were orientated on the surface of drops enclosed in a skin or membranous polymer produced by the bacteria. These droplets may represent the hydrocarbon/water emulsion of the liquid culture. The growth of A. calcoaceticus MM5 on media containing both hydrocarbon and water-soluble substrates as carbon sources also results in the formation of a film, consisting of amorphous and membranous layers. The bacteria were connected to the biofilm and showed intercellular contacts through cell-surface appendages, forming a complex network. The importance of the biofilms for bacterial adhesion to oil droplets and for its nourishment is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Asperger O, Kleber HP (1991) Metabolism of alkanes by Acinetobacter. Towner KJ, Bergogne-Bérézin E, Fewson CA (eds) The biology of Acinetobacter. Plenum, New York, pp 323–350

    Google Scholar 

  • Atlas RM, Bartha R (1992) Hydrocarbon biodegradation and oil spill bioremediation. In: Marshall KC (ed) Advances in microbial ecology. Plenum, New York London, pp 287–338

    Google Scholar 

  • Borneleit P, Hermsdorf T, Claus R, Walker P, Kleber HP (1988) Effect of hexadecane-induced vesiculation on the outer membrane of Acinetobacter calcoaceticus. J Gen Microbiol 134: 1983–1992

    Google Scholar 

  • Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–9170

    CAS  Google Scholar 

  • Bosch MP, Robert M, Mercadé ME, Espuny MJ, Parra JL, Guinea J (1988) Surface active compounds on microbial cultures. Tenside Surf Deterg 25:208–211

    Google Scholar 

  • Bouvet PJM, Grimont PAD (1986) Taxonomy of the genus Acinetobacter with the recognition of Acinetobacter baumannii sp. nov., and Acinetobacter haemolyticus sp. nov., Acinetobacter johnsonii sp. nov., and Acinetobacter junii sp. nov. and emended descriptions of Acinetobacter calcoaceticus and Acinetobacter lwoffii. Int J Syst Bacteriol 36:228–240

    Google Scholar 

  • Bushnell LD, Haas HF (1941) The utilization of certain hydrocarbons by microorganisms. J Bacteriol 41:653–673

    Google Scholar 

  • Claus R, Käppeli O, Fiechter A (1984) Possible role of extracellular membrane particles in hydrocarbon utilization by Acinetobacter calcoaceticus 69-V.J Gen Microbiol 1230:1035–1039

    Google Scholar 

  • Costerton JW, Cheng K-J, Geesey GG, Ladd TI, Nickel JC, Dasgupta M, Marrie TJ (1987) Bacterial biofilms in nature and disease. Annu Rev Microbiol 41:435–464

    Google Scholar 

  • Fiechter A (1992) Biosurfactants:moving towards industrial application. Trends Biotechnol 10:208–217

    Article  CAS  PubMed  Google Scholar 

  • Finnerty WR (1994) Biosurfactants in environmental biotechnology. Curr Opin Biotechnol 5:291–295

    Google Scholar 

  • Fletcher M (1994) Bacterial biofilms and biofolding. Curr Opin Biotechnol 5:302–306

    Google Scholar 

  • Goldman S, Shabtai Y, Rubinovitz C, Rosenberg E, Gutnich DL (1982) Emulsan in Acinetobacter calcoaceticus RAG-1; distribution of cell-free and cell-associated cross-reacting material. App Environ Microbiol 44:165–170

    Google Scholar 

  • Gutnick DL, Shabtai Y (1987) Exopolysaccharide bioemulsifiers. In: Kosaric N, Cairns WL, Gray NCC (eds) Biosurfactants and biotechnology. Dekker, New York Basel, pp 211–246

    Google Scholar 

  • Gutnick DL, Allon R, Levy C, Petter R, Minas W (1991) Applications of Acinetobacter as an industrial microorganism. In: Towner KJ, Bergogne-Bérézin E, Fewson CA (eds) The biology of Acinetobacter. Plenum, New York, pp 411–441

    Google Scholar 

  • Hanson RS, Phillips JA (1981) Chemical composition. In: Gerhardt P, Murray RGE, Costilow RN, Wester EW, Wood WA, Krieg NR, Phillips GB (eds) Manual methods for general bacteriology. American Society for Microbiology, Washington, pp 328–364

    Google Scholar 

  • Juni E (1984) Genus III. Acinetobacter Brisou et Prévot 1954. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol. I. Williams & Wilkins, Baltimore, pp 303–307

    Google Scholar 

  • Käppeli O, Fiechter A (1981) Properties of hexadecane uptake by Candida tropicalis. Curr Microbiol 6:21–26

    Google Scholar 

  • Kates M (1972) Technics of lipodology. North Holland/American Elsevier, New York

    Google Scholar 

  • Kennedy RS, Finnerty WR, Sudarsanan K, Young RA (1975) Microbial assimilation of hydrocarbons. I. The fine structure of a hydrocarbon oxidizing Acinetobacter sp. Arch Microbiol 102:75–83

    Google Scholar 

  • Marshall KC (1992) Biofilms: an overview of bacterial adhesion, activity, and control at surfaces. Am Soc Microbiol News 58:202–207

    Google Scholar 

  • Moppar R, Gindler JM (1973) A new non corrosive dye reagent to automatic sugar chromatograph. Anal Biochem 56:440–442

    Google Scholar 

  • Rosenberg E (1986) Microbial surfactants. CRC Crit Rev Microbiol 3:109–132

    Google Scholar 

  • Rosenberg E (1993) Exploiting microbial growth on hydrocarbons — new markets. Trends Biotechnol 11:419–424

    Google Scholar 

  • Rosenberg E, Legmann R, Kushmaro A, Taube R, Adler E, Ron EZ (1992) Petroleum bioremediation — a multiphase problem. Biodegradation 3:337–350

    Google Scholar 

  • Rosenberg M, Hayer EA, DeLarea J, Rosenberg E (1982) Role of thin fimbriae in adherence and growth of Acinetobacter calcoaceticus RAG-1 on hexadecane. Appl Environ Microbiol 44:929–937

    Google Scholar 

  • Sar N, Rosenberg E (1983) Emulsifier production by Acinetobacter calcoaceticus strains. Curr Microbiol 9:309–314

    Google Scholar 

  • Smith RN (1988) Bacterial extracellular polymers: a major cause of spoilage in middle distillate fuels. In: Houghton DR, Smith RN, Eggins HOW (eds) Biodeterioration 7. Elsevier, London New York, pp 256–262

    Google Scholar 

  • Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43

    CAS  PubMed  Google Scholar 

  • Towner KJ (1992) The genus Acinetobacter. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KM (eds) The prokaryotes: a handbook on the biology of bacteria. Ecophysiology, isolation, identification, applications, 2nd edition, vol 4. Springer, New York Berlin Heidelberg, pp 3137–3143

    Google Scholar 

  • Wasko MP, Bratt RP (1991) Properties of a biosurfactant produced by the fuel contaminant Ochrobactrum antropii. Int Biodeterior 27:265–273

    Google Scholar 

  • Wienzek KM, Fletcher M (1995) Bacterial adhesion to hydroxyl-and methyl-terminated alkanethiol self-assembled monolayers. J Bacteriol 177:1959–1966

    Google Scholar 

  • Zosim Z, Gutnick DL, Rosenberg E (1982) Properties of hydrocarbon-in-water emulsions stabilized by Acinetobacter RAG-1 emulsan. Biotechnol Bioeng 24:281–292

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marin, M., Pedregosa, A. & Laborda, F. Emulsifier production and microscopical study of emulsions and biofilms formed by the hydrocarbon-utilizing bacteria Acinetobacter calcoaceticus MM5. Appl Microbiol Biotechnol 44, 660–667 (1996). https://doi.org/10.1007/BF00172500

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00172500

Keywords

Navigation