Abstract
Currently, the applicability of enzymes in synthetic organic chemistry is well recognized. The field of enzyme-catalyzed organic synthesis has been further boosted by the recognition that enzymes can operate in organic solvents. The use of nonaqueous media for enzymatic conversions offers a number of advantages, like enhanced thermal stability of the enzyme, increased substrate solubility, a shift of the equilibrium in favor of synthesis over hydrolysis, and altered selectivity properties of the enzyme (1). However, the most important drawback of the use of enzymes in organic media is the reduced catalytic activity compared to aqueous conditions. Typically, this reduction in activity is two to six orders of magnitude (2). However, studies in our laboratory have revealed that (pre)treatment of the enzymes with crown ethers can enhance enzyme activities in nonaqueous organic solvents up to a level approaching the activity under aqueous conditions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Koskinen, A. M. P. and Klibanov, A. M. (eds.) (1996) Enzymatic Reactions in Organic Media. Blackie Academic & Professional, Glasgow, UK.
Dordick, J. S. (1989) Enzymatic catalysts in monophasic organic solvents. Enzyme Microb. Technol. 11, 194–211.
Pedersen, C. J. (1967) Cyclic polyethers and their complexes with metal salts. J. Am. Chem. Soc. 89, 2495–2496, 7017-7036.
Gokel, G. (1991) Crown Ethers and Cryptands. Royal Society of Chemistry, Cambridge, UK.
de Jong, F., Reinhoudt, D. N., and Smit, C. J. (1976) On the role of water in the complexation of alkylammonium salts by crown ethers. Tetrahedron Lett. 1371–1374.
Odell, B. and Earlam, G. (1985) Dissolution of proteins in organic solvents using macrocyclic polyethers: association constants of a cytochrome C-[1,2-14C2]-18-crown-6 complex in methanol. J Chem. Soc. Chem. Commun. 359–361.
Schmitke, J. L., Wescott, C. R., and Klibanov, A. M. (1996) The mechanistic dissection of the plunge in enzymatic activity upon transition from water to anhydrous solvents. J. Am. Chem. Soc. 118, 3360–3365.
Broos, J., Martin, M. N., Rouwenhorst, I., Verboom, W., and Reinhoudt, D. N. (1991) Acceletation of enzyme-catalyzed reactions in organic solvents by crown ethers. Recl. Trav. Chim. Pays-Bas 110, 222–225.
Engbersen, J. F. J., Broos, J., Verboom, W., and Reinhoudt, D. N. (1996) Effects of crown ethers and small amounts of cosolvent on the activity and enantioselectivity of α-chymotrypsin in organic solvents. Pure Appl. Chem. 68, 2171–2178.
Broos, J., Sakodinskaya, I. K., Engbersen, J. F. J., Verboom, W., and Reinhoudt, D. N. (1995) Large activation of serine proteases by pretreatment with crown ethers. J Chem. Soc. Chem. Commun. 255–256.
van Unen, D. J., Engbersen, J. F. J., and Reinhoudt, D. N. (1998) Large acceleration of α-chymotrypsin-catalyzed dipeptide formation by 18-crown-6 in organic solvents. Biotechnol. Bioeng. 59, 553–556.
van Unen, D. J., Engbersen, J. F. J., and Reinhoudt, D. N. (1998) Effects of crown ethers on the activity of enzymes in peptide formation in organic media, in Progress in Biotechnology, vol. 15: Stability and Stabilization of Biocatalysts (Ballesteros, A., Plou, F. J., Iborra, J. L., Halling, P. J., eds.), Elsevier, Amsterdam.
Zaks, A. and Klibanov, A. M. (1988) Enzymatic catalysts in nonaqueous solvents. J. Biol. Chem. 263, 3194–3201.
Dabulis, K. and Klibanov, A. M. (1993) Dramatic enhancement of enzymatic activity in organic solvents by lypoprotectants. Biotechnol. Bioeng. 41, 566–571.
Otamiri, M., Aldercreutz, P., and Mattiasson, B. (1992) Complex formation between chymotrypsin and ethyl cellulose as a means to solubilize the enzyme in active form in toluene. Biocatalysis 6, 291–305.
Halling, P. J. (1992) Salt hydrates for water activity control with biocatalysts in organic media. Biotechnol. Tech. 6, 271–276.
van Unen, D. J., Sakodinskaya, I. K., Engbersen, J. F. J., and Reinhoudt, D. N. (1998) Crown ether activation of cross-linked subtilisin Carlsberg crystals in organic solvents. J. Chem. Soc. Perkin Trans. 1, 3341–3343.
St. Clair, N. L. and Navia, M. A. (1992) Cross-linked enzyme crystals as robust biocatalysts. J. Am. Chem. Soc. 114, 7314-7316. 19. Schmid, R. D. and Verger, R. (1998) Lipases: interfacial enzymes with attractive applications. Angew. Chem. Int. Ed. 37, 1608–1633.
Gonzalez-Navarro, H. and Braco, L. (1998) Lipase-enhanced activity in flavour ester reactions by trapping enzyme conformers in the presence of interfaces. Biotechnol. Bioeng. 69, 122–127.
Xu, Z.-F., Affleck, R., Wangikar, P., Suzawa, V., Dordick, J. S., and Clark, D. S. (1994) Transition state stabilization of subtilisins in organic media. Biotechnol. Bioeng. 43, 515–520.
Broos, J., Arend, R., van Dijk, G. B., Verboom, W., Engbersen, J. F. J., and Reinhoudt, D. N. (1996) Enhancement of tyrosinase activity by macrocycles in the oxidation of p-cresol in organic solvents. J. Chem. Soc., Perkins Trans. 1, 1415–1417.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
van Unen, DJ., sEngbersen, J.F.J., Reinhoudt, D.N. (2001). The Effects of Crown Ethers on the Activity of Enzymes in Organic Solvents. In: Vulfson, E.N., Halling, P.J., Holland, H.L. (eds) Enzymes in Nonaqueous Solvents. Methods in Biotechnology, vol 15. Humana Press. https://doi.org/10.1385/1-59259-112-4:213
Download citation
DOI: https://doi.org/10.1385/1-59259-112-4:213
Publisher Name: Humana Press
Print ISBN: 978-0-89603-929-2
Online ISBN: 978-1-59259-112-1
eBook Packages: Springer Protocols