Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Genetic Construction of Lactose-Utilizing Strains of Pseudomonas Aeruginosa and Their Application in Biosurfactant Production

Bio/Technology volume 6, pages 1335–1339 (1988)Cite this article

Abstract

We inserted the E. coli lacZY genes into the chromosomes of Pseudomonas aeruginosa strains PAO-1 and PG-201 using a bicomponent transposition system, and obtained transconjugant strains with one set of lacZY genes integrated into the chromosomes at unique locations. The strains were positive in the o-nitrophenylgalacto-side (ONPG) test and produced E. coli β-galactosidase-related proteins in Western blots. The transconjugants grew well in lactose-based media (minimal medium and whey) albeit with reduced initial rates as compared to growth in glucose-based minimal media. Pseudomonas rhamnolipids were produced during stationary growth in lactose-based minimal media and whey, showing that waste products can be effectively used for important bio-technological processes.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Hisatsuka, K., Nakahara, T., Sano, N., and Yamada, K. 1971. Formation of rhamnolipid by Pseudomonas aeruginosa and its function in hydrocarbon fermentation. Agric. Biol. Chem. 35:686–692.

    Article  Google Scholar 

  2. Itoh, S., and Suzuki, T. 1972. Effect of rhamnolipids on growth of Pseudomonas aeruginosa mutant deficient in n-paraffin-utilizing ability. Agr. Biol. Chem. 36:2233–2235.

    Article  CAS  Google Scholar 

  3. Itoh, S., Honda, H., Tomita, F., and Suzuki, T. 1971. Rhamnolipids produced by Pseudomonas aeruginosa grown on n-paraffin. The Journal of Antibiotics 24:855–859.

    Article  CAS  Google Scholar 

  4. Edwards, J. and Hayashi, J. 1965. Structure of a rhamnolipid from Pseudomonas aeruginosa. Arch. Biochem. Biophys. 111:415–421.

    Article  CAS  PubMed  Google Scholar 

  5. Kosaric, N., Gray, N.C.C., and Cairns, W.L. 1983. Microbial emulsifiers and de-emulsifiers, p. 575–592. In: Biotechnology, vol. 3. Rehm, H. J. and Reed, G., (eds.) Verlag Chemie, Weinheim.

    Google Scholar 

  6. Finnerty, W.R., and Singer, M.E. 1983. Microbial enhancement of oil recovery. Bio/Technology 1:47–54.

    Google Scholar 

  7. Moulin, G. and Galzy, P. 1984. Whey, a potential substrate for biotechnology, p. 347–374. In: Biotechnology and Genetic Engineering Reviews 1. Russell, G. E. (ed.).

    Google Scholar 

  8. Drahos, D.J., Hemming, B.C., and McPherson, S. 1986. Tracking recombinant organisms in the environment: β-galactosidase as a selectable non-antibiotic marker for fluorescent pseudomonads. Bio/Technology 4:439–444.

    CAS  Google Scholar 

  9. Baumberg, S., Cornelis, G., Panagiotakopoulos, and Roberts, M. 1980. Expression of the lactose transposon Tn951 in Escherichia coli, Proteus and Pseudomonas. Journal of General Microbiology 119:257–262.

    CAS  PubMed  Google Scholar 

  10. Barry, G.F. 1986. Permanent insertion of foreign genes into the chromosome of soil bacteria. Bio/Technology 4:446–449.

    CAS  Google Scholar 

  11. Holloway, B.W. 1969. Genetics of pseudomonads. Bacteriol. Rev. 33:419–433.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Guerra Santos, L.H., Käppeli, O., and Fiechter, A. 1986. Dependence of Pseudomonas aeruginosa continuous culture biosurfactant production on nutritional and environmental factors. Applied Microbiology and Biotechnology 24:443–448.

    Article  CAS  Google Scholar 

  13. Ubben, D. and Schmitt, R. 1986. Tnl721 derivatives for transposon mutagenesis, restriction mapping and nucleotide sequence analysis. Gene 41:145–152.

    Article  CAS  PubMed  Google Scholar 

  14. Gray, M.R., Colot, H.V., Guarente, L., and Rosbash, M. 1982. Open reading frame cloning: identification, cloning, and expression of open reading frame DNA. Proc. Natl. Acad. Sci. USA 79:6598–6602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Lichtenstein, C., and Brenner, S. 1981. Site-specific properties of Tn7 transposition into the E. coli chromosome. Mol. Gen. Genet. 183:380–387.

    Article  CAS  PubMed  Google Scholar 

  16. Boyer, H.W., and Roulland-Dussoix, D. 1969. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J. Mol. Biol. 41:459–472.

    Article  CAS  PubMed  Google Scholar 

  17. Cornelis, G., Ghosal, D., and Saedler, H. 1978. Tn951: a new transposon carrying a lactose operon. Mol. Gen. Genet. 160:215–224.

    Article  CAS  PubMed  Google Scholar 

  18. Walsh, P.M., Haas, M.J., and Somkuti, G.A. 1984. Genetic construction of lactose-utilizing Xanthomonas campestris. Applied and Environ mental Microbiology 47:253–257.

    CAS  Google Scholar 

  19. Meselson, M. and Yuan, R. 1968. DNA restriction enzyme from E. coli. Nature 217:1110–1114.

    Article  CAS  PubMed  Google Scholar 

  20. Yanisch-Perron, C., Vieira, J., and Messing, J. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mp18 and pUC19 vectors. Gene 33:103–119.

    Article  CAS  PubMed  Google Scholar 

  21. Maniatis, T., Fritsch, E.F., Sambrook, J. 1982. Molecular cloning-A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  22. Marmur, J. 1961. A procedure for the isolation of deoxyribonucleic acid from microorganisms. J. Mol. Biol. 3:208–218.

    Article  CAS  Google Scholar 

  23. Feinberg, P., and Vogelstein, B. 1984. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 137:266–267.

    Article  CAS  PubMed  Google Scholar 

  24. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685.

    Article  CAS  PubMed  Google Scholar 

  25. Towbin, H., Staehelin, T., and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. USA 76:4350–4354.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Johnson, D.A., Gautsch, J.W., Sportsman, J.R., and Elder, J.H. 1984. Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal. Tech. 1:3–8.

    Article  CAS  Google Scholar 

  27. Hawkes, R., Niday, E., and Gordon, J. 1982. A dot-immunobinding assay for monoclonal and other antibodies. Analytical Biochemistry 119:142–147.

    Article  CAS  PubMed  Google Scholar 

  28. Miller, J.H. 1972. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  29. Christie, W.W. 1982. Lipid Analysis. 2nd Edition, Pereamon Press, p. 119–120.

    Google Scholar 

  30. Beutler, H.O. 1984. p. 104–112. In: Methods of Enzymatic Analysis, 3rd ed., Vol. 6. Bergmeyer, H. U. (ed.) Verlag Chemie, Weinheim, Deerfield Beach/Florida, Basel.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Biotechnologie, ETH-Hönggerberg, CH-8093, Zürich, Switzerland

    Andreas K. Koch, Jakob Reiser & Armin Fiechter

  2. BIDECO AG, CH-5401, Baden, Switzerland

    Othtnar Käppeli

Authors
  1. Andreas K. Koch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jakob Reiser
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Othtnar Käppeli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Armin Fiechter
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koch, A., Reiser, J., Käppeli, O. et al. Genetic Construction of Lactose-Utilizing Strains of Pseudomonas Aeruginosa and Their Application in Biosurfactant Production. Nat Biotechnol 6, 1335–1339 (1988). https://doi.org/10.1038/nbt1188-1335

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1188-1335

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing