Abstract
It has been shown in the last few years that enzymatic reactions can be carried out efficiently in nearly equimolar mixtures of substrates in the absence of added solvents (1,2). However, for solvent-free reactions to be efficient, a reasonable degree of miscibility between substrates is usually required. This limits the practical utility of this approach to biotransformations of substances that are either liquid at the reaction temperature (e.g., lipids [see other chapters in this volume]) or can be chemically “engineered” to become soluble in the other liquid substrate (3-9). In order to overcome this problem, we have developed a complementary bioreaction system where enzymatic transformations take place in the liquid phase of a low-melting-point eutectic mixture of substrates (10-14). Such eutectics can be formed by mixing molten substrates together at elevated temperatures and then cooling the mixture down to an ambient temperature (11). A small amount of organic solvent or water is often used to further suppress the melting point and/or to decrease the viscosity of the reaction medium (12). Enzymatic transformations in eutectic mixtures have been successfully used for the preparation of numerous peptide derivatives (10,13,14).
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References
Sarney, D. B. and Vulfson, E. N. (1995) Application of enzymes to the synthesis of surfactants. Trends Biotechnol. 13, 164–172.
Vulfson, E. N., Gill, I., and Sarney, D. B. (1995) Productivity of enzymatic catalysis in non-aqueous media: new developments, in Enzymatic Reactions in Organic Media (Klibanov, A. M., Koskinen, A. M. P., eds.), Blackie Academic and Professional, Glasgow, pp. 244–265.
Adelhorst, K., Bjorkling, F., Godtfredsen, S. E., and Kirk, O. (1990) Enzyme catalyzed preparation of 6-O-acyl-glucopyranosides. Synthesis 2, 112–115.
Fregapane, G., Sarney, D. B., and Vulfson, E. N. (1991) Enzymic solvent-free synthesis of sugar acetal fatty acid esters. Enzyme Microb. Technol. 13, 796–800.
Fregapane, G., Sarney, D. B., Greenberg, S., Knight, D. J., and Vulfson, E. N. (1994) Enzymatic synthesis of monosaccharide fatty acid esters and their comparison with conventional products. J. Am. Oil Chem. Soc. 71, 87–91.
Sarney, D. B., Fregapane, G., and Vulfson, E. N. (1994) Facile chemo-enzymatic synthesis of monosaccharide fatty acid esters. Biocatalysis 11, 9–18.
Sarney, D. B., Kappeler, H., Fregapane, G., and Vulfson, E. N. (1994) Chemo-enzymatic synthesis of disaccharide fatty acid esters. J. Am. Oil Chem. Soc. 71, 711–714.
Sarney, D. B., Barnard, M. J., MacManus, D. A., and Vulfson, E. N. (1996) Application of lipases to the regioselective synthesis of sucrose fatty acid monoesters. J. Am. Oil Chem. Soc. 73, 1481–1487.
Valivety, R., Gill, I., and Vulfson, E. N. (1997) Enzymatic synthesis of novel bola-and gemini-surfactants. J. Surfact. Deterg. 1, 177–185.
Gill, I. and Vulfson, E. N. (1993) Enzymatic synthesis of short peptides in heterogeneous mixtures of substrates. J. Am. Chem. Soc. 115, 3348,3349.
Gill, I. and Vulfson, E. N. (1994) Enzymatic catalysis in heterogeneous eutectic mixtures of substrates. Trends Biotechnol. 12, 118–122.
Gill, I., Lopez-Fandino, R., and Vulfson, E. N. (1994) Protease-catalysed synthesis of oligopeptides in heterogeneous substrate mixtures. Biotechnol. Bioeng. 43, 1024–1030.
Lopez-Fandino, R., Gill, I., and Vulfson, E. N. (1994) Enzymatic catalysis in heterogeneous mixtures of substrates: the role of the liquid phase and the effects of adjuvants. Biotechnol. Bioeng. 43, 1016–1023.
Jorba, X., Gill, I., and Vulfson, E. N. (1995) Enzymatic synthesis of the “delicious peptide” fragments in eutectic mixtures J. Agric. Food Chem. 73, 2536–2541.
Nilsson, K. G. I. (1988) Enzymatic synthesis of oligosaccharides. Trends Biotechnol. 6, 256–264.
Wang, P. G., Fitz, W., and Wong, C.-H. (1995) Making complex carbohydrates via enzymatic routes. CHEMTECH 25, 22–32.
Fernandez-Mayoralas, A. (1997) Synthesis and modification of carbohydrates using glycosidases and lipases. Topics Curr. Chem. 186, 1–20.
Millqvist-Fureby, A., Gill, I. S., and Vulfson, E. N. (1998) Enzymatic transformations in supersaturated solutions: I. A general study with glycosidases. Biotechnol. Bioeng. 60, 190–196.
Millqvist-Fureby, A., MacManus, D. A., Davies, S., and Vulfson, E. N. (1998) Enzymatic transformations in supersaturated solutions: II Synthesis of disaccha-rides via transglycosylation. Biotechnol. Bioeng. 60, 197–203.
Millqvist-Fureby, A., Gao, C., and Vulfson, E. N. (1998) Enzymatic synthesis of ethoxylated glycoside esters using glycosidases in supersaturated solutions and lipases in organic solvents. Biotechnol. Bioeng. 59, 747–753.
MacManus, D. A. and Vulfson, E. N. (2000) Regioselectivity of enzymatic glycosylation of 6-O-acyl-glycosides in supersaturated solutions. Biotechnol. Bioeng., in press.
Franks, F. (1993) Solid aqueous solutions. Pure Appl. Chem. 65, 2527–2537.
Slade, L. and Levine, H. (1988) Non-equilibrium behavior of small carbohydrates-water systems. Pure Appl. Chem. 12, 1841–1864.
Orford, P. D., Parker, R., and Ring, S. G. (1990) Aspects of glass transition behaviour of mixtures of carbohydrates of low molecular weight. Carbohydr. Res. 196, 11–18.
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MacManus, D.A., Millqvist-Fureby, A., Vulfson, E.N. (2001). Biotransformations in Supersaturated Solutions. 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/10.1385/1-59259-112-4:545
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DOI: https://doi.org/10.1385/1-59259-112-4:545
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