Skip to main content
SpringerLink
Log in

Liquid-liquid extraction of a recombinant cutinase from fermentation media with AOT reversed micelles

  • Originals
  • Published:
Bioprocess Engineering Aims and scope Submit manuscript

Abstract

This work reports the extraction and back-extraction of an intracellular recombinant cutinase from complex biological media using AOT reversed micelles in isooctane. Cutinase was recovered from different complex media namely, fermentation broths and supernatants after cell disruption by osmotic shock and sonication. The application of the AOT reversed micellar system to the extraction of cutinase allowed activity yields and purification factors ranging from about 5% to 50% and 1.2 to 10.2, respectively, depending on the biological medium.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Taipa, M.A.; Aires-Barros, MR.; Cabral, J.M.S.: Purification of lipases. J. Biotechnol. 26 (1992) 111–142

    Google Scholar 

  2. Aires-Barros, M.R.; Cabral, J.M.S.: Liquid-liquid extraction In: J.F. Kennedy and J.M.S. Cabral (Eds.): Recovery Processes for Biological Materials, Chichester: John Wiley & Sons Ltd. 1993 223–246

    Google Scholar 

  3. Cabral, J.M.S.; Aires-Barros, M.R.: Reversed micelles in liquid-liquid extraction In: J.F. Kennedy and J.M.S. Cabral (Eds.): Recovery Processes for Biological Materials, Chichester: John Wiley & Sons Ltd. (1993) 247–271.

    Google Scholar 

  4. Dekker, M.; Hilhorst, R.; Laane, C.: Isolating enzymes by reversed micelles. Anal. Biochem. 178 (1989) 217–226

    Google Scholar 

  5. Hatton, T.A.: Reversed micellar extraction of proteins In: J.F. Scamehorn and J.H. Horwell (Eds.): Surfactant-Based Separation Process, vol. 33, New York (1989) 55–90.

  6. Castro, M.J.M.; e Cabral, J.M.S.: Reversed micelles in biotechnological proceses. Biotech. Adv. 6 (1988) 151–167

    Google Scholar 

  7. Rahaman, R.S.; Chee, J.Y.; Cabral, J.M.S.; Hatton, T.A.: Recovery of an extracellular alkaline protease from whole fermentation broth using reversed micelles. Biotechnol. Progr. 4 (1988) 218–224

    Google Scholar 

  8. Krei, G.A.; Hustedt, H.: Extraction of enzymes by reversed micelles. Chem. Eng. Sci. 47 (1992) 99–111

    Google Scholar 

  9. Martinez, C.; De Geus, P.; Lauwereys, M.; Matthyssens, G.; Cambillau, C.: Fusarium solani cutinase is a lipolytic enzyme with a catalytic serine accessible to solvent. Nature 356 (1992) 615–681

    Google Scholar 

  10. Lowry, R.R.; Tinsley, J.I.: Rapid calorimetric determination of free fatty acids. J. Am. Oil Chem. Soc. 53 (1976) 470–472

    Google Scholar 

  11. Smith, P.K.; Krohn, R.I.; Hermanson, G.T.; Mallia, A.K.; Gardner, F.H.; Provenzano, M.D.; Fujimoto, C.K.; Goeke, N.M.; Olson, B.J.; Klenk, D.C.: Measurement of protein using bicinchoninic acid. Anal Biochem. 150 (1985) 76–85

    Google Scholar 

  12. Medeiros, G.M.M.; Leitão, M.F.; Leitão, S.M.B.: Costa Fluorescence of acridine and acridine 9-carboxylic acid in anionic micelles. J. Photochem. Photobiol. A: Chem. 72 (1993) 225–233

    Google Scholar 

  13. Lauwereys, M.; De Geus, P.; De Meutter, J.; Stanssens, P.; Matthyssens, G.: Cloning, expression and characterization of cutinase, a fungal lipolytic enzyme In: L. Alberghina, R.D. Schmid and R. Verger (Eds.): Lipases: Structure, Mechanism and Genetic Engineering), GBF Monographs, vol 16, (1993) 243–251

  14. Carneiro-da-Cunha, M.G.; Cabral, J.M.S.; Aires-Barros, M.R.: Studies on the extraction and back-extraction of a recombinant cutinase in a reversed micellar extraction process. Bioprocess Eng. 11 (1994) 203–208

    Google Scholar 

  15. Melo, E.P.; Costa, S.M.B.; Cabral, J.M.S.: Structural effect of reversed micelles of AOT over a recombinant cutinase from Fusarium solani pisi: A study state fluorescence study. Ann. N.Y. Acad. Sci. 750 (1995) 85–88

    Google Scholar 

  16. Soliday, C.L.; Flurkey, W.H.; Okita, T.W.; Kolattukudy, P.E.: Cloning and structure determination of cDNA for cutinases, an enzyme involved in fungal penetration of plants. Proc. Natl. Acad. Sci. USA 81 (1984) 3939–3943

    Google Scholar 

  17. Weinryb, I.; Steiner, R.F.: The luminescence of the aromatic amino acids In: R.F. Steiner and I. Weinryb (Eds.): Excitated States of Proteins and Nucleic Acids New York, Plenum Press, (1971) 277–318

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratório de Engenharia Bioquímica, Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1000, Lisboa, Portugal

    M. G. Carneiro-da-Cunha, E. P. Melo, J. M. S. Cabral & M. R. Aires-Barros

Authors
  1. M. G. Carneiro-da-Cunha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. P. Melo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. M. S. Cabral
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. R. Aires-Barros
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Maria das Graças Carneiro da Cunha, from ITEP-Instituto Tecnológico do Estado de Pernambuco, acknowledges a Ph.D fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Centro de Pesquisa Aggeu Magalhães, Recife — PE — Brasil. E. P. Melo thanks Junta Nacional de Investigação Científica, Lisboa, Portugal, for providing a Ph.D. fellowship. The scientific support given by Prof. Sílvia M. B. Costa for the spectroscopic data and further discussions are particularly acknowledged.

This work was partly financed by the BRIDGE and BIOTECHNOLOGY Programmes (Contracts BIOT-CT91-0274(DTEE) and BIOT 2 CT-943016).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Carneiro-da-Cunha, M.G., Melo, E.P., Cabral, J.M.S. et al. Liquid-liquid extraction of a recombinant cutinase from fermentation media with AOT reversed micelles. Bioprocess Engineering 15, 151–157 (1996). https://doi.org/10.1007/BF00369619

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00369619

Keywords

Search

Navigation