Synthesis of Esters Catalyzed by Lipases in Water-in-Oil Microemulsions

  • Haralambos Stamatis
  • Aristotelis Xenakis
  • Fragiskos N. Kolisis
Part of the Methods in Biotechnology book series (MIBT, volume 15)


There are two basic advantages in using enzymes as catalysts in organic media instead of aqueous solutions. First, organic solvents favor the solubility of hydrophobic substrates and, second, the presence of such solvents shifts the thermodynamic equilibrium of condensation/hydrolysis reactions in favor of the desired product. Different approaches have been proposed to facilitate the reversal of the normal hydrolytic action of enzymes. These include various macroheterogeneous biphasic systems such as liquid-liquid systems composed of a water-immiscible organic solvent and water, nearly anhydrous systems in which the enzyme is usually suspended as a powder or in an immobilized form adsorbed onto a suitable carrier in organic solvents or gases in a supercritical state, and various homogeneous and microheterogeneous media such as mixtures of water-miscible organic solvent and water as well as different types of microemulsion system (reverse micelles). The subject of enzyme catalysis in media with low water content has been reviewed by several authors (1-6).


Surfactant Molecule Reverse Micelle Hydrophobic Substrate Microemulsion System Sulfosuccinate Sodium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Ballesteros, A., Bornscheuer, U., Capewell, A., Combes, D., Condoret, J.-S., Koening, K., et al. (1995) Review article: enzymes in nonconventional phases. Biocatal. Biotransform. 13, 1–42.CrossRefGoogle Scholar
  2. 2.
    Dodrick, J. S. (1989) Enzymatic catalysis in monophasic organic solvent. Enzyme Microb. Technol. 11, 194–211.CrossRefGoogle Scholar
  3. 3.
    Khmelnitsky, Y. L., Levashov, A. V., Klyachko, N. L., and Martinek K. (1988) Engineering biocatalytic systems in organic media with low water content. Enzyme Microb. Technol. 10, 710–724.CrossRefGoogle Scholar
  4. 4.
    Klibanov, A. M. (1989) Enzymatic catalysis in anhydrous organic solvents. Trends Biochem. Sci. 14, 141–144.CrossRefGoogle Scholar
  5. 5.
    Stamatis, H., Xenakis, A. and Kolisis, F. N. (1999) Bioorganic reactions in microemulsions: the case of lipase. Biotechnol. Adv., in press.Google Scholar
  6. 6.
    Stamatis, H., Xenakis, A., Provelegiou M., and Kolisis, F. N. (1993) Esterification reactions catalyzed by lipases in microemulsions. The role of enzyme localization in relation to its selectivity. Biotechnol. Bioeng. 42, 103–110.CrossRefGoogle Scholar
  7. 7.
    Eicke, H. F. and Rehak, J. (1976) On the formation of water/oil microemulsions. Helv. Chim. Acta 59, 2883–2891.CrossRefGoogle Scholar
  8. 8.
    Luisi, P. L. and Magid, L. (1986) Solubilization of enzymes and nucleic acids in hydrocarbon micellar solutions. CRC Crit. Rev. Biochem. 20, 409–474.CrossRefGoogle Scholar
  9. 9.
    Martinek, K., Levashov, A. V., Klyachko, N. L., Khmelnitsky, Y. L., and Berezin, I. V. (1986) Micellar enzymology. Eur. J. Biochem. 155, 453–468.CrossRefGoogle Scholar
  10. 10.
    Maestro, M. (1989) Enzymatic activity in reverse micelles—some modellistic considerations on bell-shaped curves. J. Mol. Liquids 42, 71–82.CrossRefGoogle Scholar
  11. 11.
    Verhaert, R. M. D. and Hilhorst, R. (1991) Enzymes in reverse micelles: 4. Theoretical analysis of a one-substrate/one-product conversion and suggestions for efficient application. Recl. Trav. Chim. Pays-Bas 110, 236–246.CrossRefGoogle Scholar
  12. 12.
    Sanchez-Ferrer, A., Perez-Gilabert, M., and Garcia-Carmona, F. (1992) Proteininterface interactions in reverse micelles, Biocatalysis in Non-Conventional Media, Progress in Biotechnology vol. 8 (Tramper, J., Vermue, M. H., Beeftink, H. H., and von Stockar, U., eds.), Elsevier, Amsterdam, pp. 181–188.Google Scholar
  13. 13.
    Otero, C., Rua, M. L., and Robledo, L. (1995) Influence of the hydrophobicity of lipase isoenzymes from Candida rugosa on its hydrolytic activity in reverse micelles. FEBS Lett. 360, 202–206.CrossRefGoogle Scholar
  14. 14.
    Luithi, P. and Luisi, P. L. (1984) Enzymatic synthesis of hydrocarbons—soluble peptides with reverse micelles. J. Am. Chem. Soc. 106, 7285,7286.Google Scholar
  15. 15.
    Hayes, D. G. and Gulari, E. (1990) Esterification reactions of lipase in reverse micelles. Biotechnol. Bioeng. 35, 793–801.CrossRefGoogle Scholar
  16. 16.
    Smolders, A. J. J., Pinheiro, H. M., Noronha, P., and Cabral, J. M. S. (1991) Steroid bioconversion in a microemulsion system. Biotechnol. Bioeng. 38, 1210–1217.CrossRefGoogle Scholar
  17. 17.
    Hilhorst, R., Spruijt, R., Laane, C., and Verger, C. (1984) Rules for the regulation of enzyme activity in reversed micelles as illustrated by the conversion of apolar steroids by 20-b-hydroxysteroid dehydrogenase. Eur. J. Biochem. 144, 459–462.CrossRefGoogle Scholar
  18. 18.
    Larsson, K. M., Adlercreutz, P., and Mattiasson, B. (1990) Ezymatic catalysis in microemulsions: enzyme reuse and product recovery. Biotechnol. Bioeng. 36, 135–141.CrossRefGoogle Scholar
  19. 19.
    Stamatis, H., Xenakis, A., and Kolisis, F. N. (1995) Studies on enzyme reuse and product recovery in lipase catalyzed-reactions in microemulsions. Ann. NYAcad. Sci. 750, 237–241.CrossRefGoogle Scholar
  20. 20.
    Stamatis, H., Xenakis, A., Dimitriadis, E., and Kolisis, F. N. (1995) Catalytic behavior of Pseudomonas cepacia lipase in w/o microemulsions. Biotechnol. Bioeng. 45, 33–41.CrossRefGoogle Scholar
  21. 21.
    Stamatis, H., Macris, J., and Kolisis, F. N. (1996) Esterification of hydrophilic diols catalysed by lipases in microemulsions. Biotechnol. Lett. 18, 541–546.CrossRefGoogle Scholar
  22. 22.
    Stamatis, H. (1996) Enzymatic modification of amphiphilic substrates in microemulsions. Ph.D. thesis, University of Patras, Greece.Google Scholar

Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Haralambos Stamatis
    • 1
  • Aristotelis Xenakis
    • 2
  • Fragiskos N. Kolisis
    • 2
  1. 1.Department of Chemical EngineeringNational Technical UniversityAthensGreece
  2. 2.Industrial Enzymology UnitInstitute of Biological Research and Biotechnology, The National Hellenic Research FoundationAthensGreece

Personalised recommendations