Skip to main content
SpringerLink
Log in

The use of reverse micelles for the separation of proteins

  • Chapter
Highly Selective Separations in Biotechnology

Abstract

Liquid-liquid extraction technology has been successfully used in the separation of compounds in chemical, petrochemical, or hydrometallurgical industries for many years. It is only recently, however, that liquid-liquid extraction technology has been recognized as potentially useful in biotechnology, namely in the purification procedure of biomolecules, such as proteins (Kadam, 1986; Abbott and Hatton, 1988). Two classes of two-phase extraction system are suitable for protein recovery: (a) biphasic aqueous polymer systems, and (b) systems in which an organic, reverse micellar solution is in equilibrium with a conjugated aqueous phase. Both systems are based on the differential partitioning of protein(s) between the immiscible feed and solvent phase to effect the extraction of the desired protein from the complex production mixture. However, the principles of solubilization greatly differ. In the following we will describe the micellar extraction system in more detail and show its usefulness in the selective extraction of proteins from crude mixtures.

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

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Abbott, N.L. and Hatton, T.A. (1988) Liquid-liquid extraction for protein separations. Chem. Eng. Progr. (August), 31–41.

    Google Scholar 

  • Aires-Barros, M.R. and Cabrai, J.M.S. (1991) Selective separation and purification of two lipases from Chromobacter viscosum using AOT reversed micelles. Biotechnol. Bioeng., 38, 1302–1307.

    Article  PubMed  CAS  Google Scholar 

  • Armstrong, D.W. and Li, W. (1988) Highly selective protein separations with reverse micellar liquid membranes. Anal. Chem., 60, 86–88.

    Article  PubMed  CAS  Google Scholar 

  • Aveyard, R., Binks, B.P. et al. (1986a) Interfacial tension minima in oil-water-surfactant systems. J. Chem. Soc., Faraday Trans. I, 82, 125–142.

    Article  CAS  Google Scholar 

  • Aveyard, R., Binks, B.P. et al. (1986b) Interfacial tension minima in oil-water-surfactant systems. J. Soc. Chem., Faraday Trans. I, 82, 1755–1770.

    Article  CAS  Google Scholar 

  • Bratko, B., Luzar, A. et al. (1988) Electrostatic model for protein/reverse micelle complexation. J Chem. Phys., 89, 545–550.

    Article  CAS  Google Scholar 

  • Brochette, P., Petit, C. et al. (1988) Cytochrome c in bis(2-ethylhexyl) sulfosuccinate reverse micelles: Structure and reactivity. J. Phys. Chem., 92, 3505–3511.

    Article  CAS  Google Scholar 

  • Bruno, P., Caselli, M. et al. (1990) A simplified thermodynamic model for protein uptake by reverse micelles: Theoretical and experimental results. J. Phys. Chem., 94, 5908–5917.

    Article  CAS  Google Scholar 

  • Carlson, A. and Nagarajan, R. (1992) Release and recovery of porcine pepsin and bovine chymosin from reverse micelles: A new technique based on isopropyl alcohol addition. Biotechnol. Progr., 8, 85–90.

    Article  CAS  Google Scholar 

  • Caselli, M., Luisi, P.L. et al. (1988) Thermodynamics of the uptake of proteins by reverse micelles: A first approximation model. J. Phys. Chem., 92, 3899–3905.

    Article  CAS  Google Scholar 

  • Coughlin, R.W. and Baclaski, J.B. (1990) N-Laurylbiotinamide as affinity surfactant. Biotechnol. Progr., 6, 307–309.

    Article  CAS  Google Scholar 

  • Dahuron, L. and Cussler, E.L. (1988) Protein extraction with hollow fibers. Am. Inst. Chem. Eng. J., 34, 130–136.

    Article  CAS  Google Scholar 

  • Dekker, M. (1993) Reversed micelles for protein purification. In Molecular interactions in Bioseparation (Ngo, T., ed.), Plenum, New York, pp. 533–544.

    Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1986) Enzyme recovery by Liquid-liquid extraction using reversed micelles. Chem. Eng. J., 33, B27–B33.

    Google Scholar 

  • Dekker, M., Baltussen, J.W.A. et al. (1987a) Reversed micellar extraction of enzymes; effect of nonionic surfactant on the distribution and extraction efficiency of α-amylase. In Biocatalysis in Organic Media (Laane, C., Tramper, J., and Lilly, M.D. eds). Elsevier, Amsterdam, pp. 285–288.

    Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1987b) Membrane based Liquid-liquid extraction of enzymes using reversed micelles. In Proc. ICOM, Tokyo, pp. 793–794.

    Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1987c) Reversed micellar extraction of enzymes: investigations on the distribution behaviour and extraction efficiency of α-amylase. In Proc. 4th Eur. Congr. Biotechnol, Amsterdam, Vol. II, pp. 507–510.

    Google Scholar 

  • Dekker, M., Hilhorst, R. et al. (1989a) Isolating enzymes by reverse micelles. Anal Biochem., 178, 217–226.

    Article  PubMed  CAS  Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1989b) Modeling and optimization of the reversed micellar extraction of α-amylase. Am. Inst. Chem. Eng. J., 35, 321–324.

    Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1990) Mass transfer rate of protein extraction with reversed micelles. Chem. Eng. Sci., 49, 2949–2957.

    Google Scholar 

  • Dekker, M., Koenen, P.H.M. et al. (1991a) Reversed micellar-membrane-extraction of enzymes. Trans. Inst. Chem. Eng., 68(C), 54–58.

    Google Scholar 

  • Dekker, M., Van’t Riet, K. et al. (1991b) Effect of temperature on the reversed micellar extraction of enzymes. Chem. Eng. J., 46, B69–B74.

    Google Scholar 

  • Dungan, S.R., Bausch, T. et al. (1991) Interfacial transport processes in the reversed micellar extraction of proteins. J. Colloid Interface Sci., 145, 33.

    Article  CAS  Google Scholar 

  • Eicke, H.-F. (1980) Surfactants in nonpolar solvents: Aggregation and micellization. Top. Curr. Chem., 87, 85–145.

    Article  PubMed  CAS  Google Scholar 

  • Fletcher, P.D.I, and Parrott, D. (1988) The partitioning of proteins between water-in-oil microemulsions and conjugate aqueous phase. J. Chem. Soc., Faraday Trans. I, 84, 1131–1144.

    Article  CAS  Google Scholar 

  • Fraaije, J.G.E.M., Rijnierse, E.-J. et al. (1990) Protein partition and ion copartition in aqueous-apolar two-liquid-phase systems. Colloid Polym. Sci., 268, 855–863.

    Article  CAS  Google Scholar 

  • Giovenco, S., Verheggen, F. et al. (1987a) Purification of intracellular enzymes from whole bacterial cells using reversed micelles. Enzyme Microb. Technol., 9, 470–473.

    Article  Google Scholar 

  • Giovenco, S., Laane, C. et al. (1987b) Reversed micelles as a bioseparation tool. In Proc. 4th Eur. Congr. Biotechnol., Amsterdam, Vol. II, pp. 503–506.

    Google Scholar 

  • Göklen, K.E. and Hatton, T.A. (1987) Liquid-liquid extraction of low molecular-weight proteins by selective solubilization in reverse micelles. Sep. Sci. Technol., 22, 831–841.

    Article  Google Scholar 

  • Goto, M., Kondo, K. et al. (1992) Protein extraction by reversed micelles using new surfactants. In Solvent Extraction (Sekine, T., ed.). Elsevier, pp. 1845–1850.

    Google Scholar 

  • Haering, G., Pessina, A. et al. (1987) Solubilization of bacterial cells in organic solvents via reverse micelles and microemulsions. Ann. N. Y. Acad. Sci., 506, 337–344.

    Article  PubMed  CAS  Google Scholar 

  • Hagen, A.J., Hatton, T.A., et al. (1990b) Protein refolding in reversed micelles: Interactions of the protein with micelle components. Biotechnol. Bioeng., 35, 966–975.

    Article  PubMed  CAS  Google Scholar 

  • Hagen, A.J., Hatton, T.A. et al. (1990a) Protein refolding in reversed micelles. Biotechnol. Bioeng., 35, 955–965.

    Article  PubMed  CAS  Google Scholar 

  • Hatton, T.A. (1989) Reversed micellar extraction of proteins. In Surfactant-Based Separation Processes (Scamehorn J.F. and Harwell, J.H., eds). Marcel Dekker, New York, Basel, pp. 55–90.

    Google Scholar 

  • Hilhorst, R., Fijneman, P. et al. (1992) Protein extraction using reverse micelles. Pure Appl. Chem., 64, 1765–1770.

    Article  CAS  Google Scholar 

  • Hochköppler, A., Pfammatter, N. et al. (1989) Activity of yeast cells solubilized in a water-in-oil microemulsion. Chimia, 43, 348–350.

    Google Scholar 

  • Hou, M.J. and Shah, D.O. (1987) Effects of the molecular structure of the interface and continuous phase on solubilization of water in water/oil microemulsions. Langmuir, 3, 1086–1096.

    Article  CAS  Google Scholar 

  • Ichikawa, S., Imai, M. et al. (1992) Solubilizing water involved in protein extraction using reversed micelles. Biotechnol. Bioeng., 39, 20–26.

    Article  PubMed  CAS  Google Scholar 

  • Israelachvili, J.N., Mitchell, D.J. et al. (1976) Theory of self-assembly of hydrocarbon amphiphiles into micelles and bilayers. J. Chem. Soc, Faraday Trans. II, 12, 1525–1568.

    Article  Google Scholar 

  • Jolivalt, C., Minier, M. et al. (1990) Extraction of α-chymotrypsin using reverse micelles. J. Colloid Interface Sci., 135, 85–96.

    Article  CAS  Google Scholar 

  • Kadam, K.L. (1986) Reversemicelles as a bioseparation tool. Enzyme Microb. Technol., 8, 266–273.

    Article  CAS  Google Scholar 

  • Klyachko, N.L., Levashov, A.V. et al. (1985) Catalysis by enzymes incorporated in reverse micelles of surface-active substances in organic solvents. Molekulyarnaya Biologiya (Engl. edn), 18, 830–840.

    Google Scholar 

  • Krei, G.A. and Hustedt, H. (1992) Extraction of enzymes by reverse micelles. Chem. Eng. Sci., 47, 99–111.

    Article  CAS  Google Scholar 

  • Lang, J., Jada, A. et al. (1988) Structure and dynamics of water-in-oil droplets stabilized by sodium bis-(2-ethylhexyl) sulfosuccinate. J. Phys. Chem., 92, 1946–1953.

    Article  CAS  Google Scholar 

  • Langevin, D. (1992) Micelles and microemulsions. Ann. Rev. Phys. Chem., 43, 341–369.

    Article  CAS  Google Scholar 

  • Leodidis, E.B. and Hatton, T.A. (1989) Specific ion effects in electrical double layers: Selective solubilization of cations in aerosol-OT reversed micelles. Langmuir, 5, 741–753.

    Article  CAS  Google Scholar 

  • Leser, M.E. and Luisi, P.L. (1990) Application of reverse micelles for the extraction of amino acids and proteins. Chimia, 44, 270–282.

    CAS  Google Scholar 

  • Leser, M.E., Luisi, P.L. et al. (1989) The use of reverse micelles for the simultaneous extraction of oil and proteins from vegetable meal. Biotechnol. Bioeng., 34, 1140–1146.

    Article  PubMed  CAS  Google Scholar 

  • Leser, M.E., Mrkoci, K. et al. (1993) Reverse micelles in protein separation: The use of silica for the back-transfer process. Biotechnol. Bioeng., 41, 489–492.

    Article  PubMed  CAS  Google Scholar 

  •  Levashov, A.V., Klyachko, N.L. et al. (1987) Superactivity of acid phosphatase in inverted micelles of surfactants in organic solvents. Dokl. Akad. Nauk SSSR (Engl, edn), 289, 279–281.

    Google Scholar 

  • Luisi, P.L., and Laane, C. (1986) Solubilization of enzymes in apolar solvents via reverse micelles. Trends Biotechnol., 4,153–161.

    Article  CAS  Google Scholar 

  • Luisi, P.L. and Magid, L.J. (1986) Solubilization of enzymes and nucleic acids in hydrocarbon micellar solutions. CRC Crit. Rev. Biochem., 20, 409–474.

    Article  PubMed  CAS  Google Scholar 

  • Luisi, P.L., Bonner, F.J. et al. (1979) Micellar solubilization of proteins in aprotic solvents and their spectroscopic characterization. Helv. Chim. Acta, 62, 740–753.

    Article  CAS  Google Scholar 

  • Luisi, P.L., Giomini, M. et al. (1988) Reverse micelles as hosts for proteins and small molecules. Biochim. Biophys. Acta, 947, 209–246.

    Article  PubMed  CAS  Google Scholar 

  • Maestro, M. and Luisi, P.L. (1986) A simplified thermodynamic model for protein uptake by reverse micelles. Presented at the 6th International Symposium on Surfactants in Solution, New Delhi, 1986; In Surfactants in Solution (1989) Vol. 9 (Mittal, K.L., ed.). Plenum Press, New York, pp. 25–45.

    Google Scholar 

  • Mao, Q. and Walde, P. (1991) Substrate effects on the enzymatic activity of alpha-chymotrypsin in reverse micelles. Biochem. Biophys. Res. Commun., 178, 1105–1112.

    Article  PubMed  CAS  Google Scholar 

  • Marcozzi, G., Correa, N. et al. (1991) Protein extraction by reverse micelles: A study of the factors affecting the forward and backward transfer of α-chymotrypsin and its activity. Biotechnol. Bioeng., 38, 1239–1246.

    Article  PubMed  CAS  Google Scholar 

  • Martinek, K., Levashov, A.V. et al. (1978) Catalysis by water-soluble enzymes in organic solvents. Stabilization of enzymes against denaturation (inactivation) through their inclusion in reversed micelles of surfactant. Dokl. Akad. Nauk. SSSR (Engl, edn), 236, 951–953.

    Google Scholar 

  • Martinek, K., Klyachko, N.L. et al. (1989) Micellar enzymology: Its relation to membranology. Biochim. Biophys. Acta, 981, 161–172.

    Article  PubMed  CAS  Google Scholar 

  • Menger, F.M. and Yamada, K. (1979) Enzyme catalysis in water pools. J. Amer. Chem. Soc., 101, 6731–6734.

    Article  CAS  Google Scholar 

  • Nitsch, W. and Plucinski, P. (1990) Two-phase kinetics of the solubilization in reverse micelles. J. Colloid Interface Sci., 136, 338–351.

    Article  CAS  Google Scholar 

  • Paradkar, V.M. and Dordick, J.S. (1991) Purification of glycoproteins by selective transport using concanavalin-mediated reverse micellar extraction. Biotechnol Progr., 1, 330–334.

    Article  Google Scholar 

  • Pfammatter, N., Guadalupe, A.A. et al. (1989) Solubilization and activity of yeast cells in water-in-oil microemulsion. Biochem. Biophys. Res. Commun., 161, 1244–1251.

    Article  PubMed  CAS  Google Scholar 

  • Plucinski, P. and Nitsch, W. (1989) Two-phase kinetics of the solubilization in reverse micelles—extraction of lysozyme. Ber. Bunsenges. Phys. Chem., 93, 994–997.

    Article  CAS  Google Scholar 

  • Plucinski, P. and Nitsch, W. (1992) Kinetics of the interfacial ion exchange in Winsor II microemulsion systems. J. Colloid Interface Sci., 154, 104–112.

    Article  CAS  Google Scholar 

  • Rahaman, S.R., Chee, J.Y. et al. (1988) Recovery of an extracellular alkaline protease from whole fermentation broth using reversed micelles. Biotechnol. Progr., 4, 218–224.

    Article  CAS  Google Scholar 

  • Ricka, J., Borkovec, M. et al. (1991) Coated droplet model of microemulsions: Optical matching and polydispersity. J. Chem. Phys., 94, 8503–8509.

    Article  CAS  Google Scholar 

  • Scholtens, B.J.R., Bruin, S. et al. (1979) Liquid-liquid mass transfer and its retardation by macro-molecular adsorption. Chem. Eng. Sci., 34, 661–670.

    Article  CAS  Google Scholar 

  • Shield, J.W., Ferguson, H.D. et al. (1986) Enzymes in reversed micelles as catalysts for organic-phase synthesis reactions. Ind. Eng. Chem. Fundam., 25, 603–612.

    Article  CAS  Google Scholar 

  • Shinoda, K. and Lindoman, B. (1987) Organized surfactant systems: microemulsions. Langmuir, 3, 135–149.

    Article  CAS  Google Scholar 

  • Weir, M.P. and Sparks, J. (1987) Purification and renaturation of recombinant human inter-leukin-2. Biochem. J., 245, 85–91.

    PubMed  CAS  Google Scholar 

  • Wolbert, R.B.G., Hilhorst, R. et al. (1989) Protein transfer from an aqueous phase into reversed micelles; the effect of protein size and charge distribution. Eur. J. Biochem., 184, 627–633.

    Article  PubMed  CAS  Google Scholar 

  • Woll, J.M. and Hatton, T.A. (1989) A simple phenomenological thermodynamic model for protein partitioning in reversed micellar systems. Bioprocess Eng., 4, 193–199.

    Article  Google Scholar 

  • Woll, J., Hatton, T.A. et al. (1989) Bioafïïnity separations using reversed micellar extraction. Biotechnol. Progr., 5, 57–62.

    Article  CAS  Google Scholar 

  • Zulauf, M. and Eicke, H.-F. (1979) Inverted micelles and microemulsions in the ternary system H2O/Aerosol-OT/iso-octane as studied by photon correlation spectroscopy. J. Phys. Chem., 83, 480–486.

    Article  CAS  Google Scholar 

Download references

Authors
  1. M. Dekker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. E. Leser
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. School of Science & Technology, University of Teesside, Middlesbrough, UK

    G. Street

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Springer Science+Business Media Dordrecht

About this chapter

Cite this chapter

Dekker, M., Leser, M.E. (1994). The use of reverse micelles for the separation of proteins. In: Street, G. (eds) Highly Selective Separations in Biotechnology. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-1322-9_5

Download citation

  • DOI: https://doi.org/10.1007/978-94-011-1322-9_5

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-4576-6

  • Online ISBN: 978-94-011-1322-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation