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Note: The numbers express the amount of detergent necessary to desorb 100% of lipase from the support.

Note: The numbers express the amount of detergent necessary to desorb 100% of lipase from the support.

evant conditions, it is convenient that the enzyme can, after enzyme deactivation, be desorbed from the support in order to reload the support with fresh enzyme and take advantage of the reversibility of the technique. Table 1 shows that the adsorbed lipases on octadecyl-Sepabeads or octyl-agarose could be fully desorbed by using moderate concentrations of detergent. Moreover, the incubations with urea and guanidine enable the full desorption of the lipase.

The regenerated supports could be used to immobilize new batches of lipase without significant differences in terms of loading capacity compared to the first immobilization.

4. Notes

1. To take suspension the tips have to be cut.

2. In some instances, if immobilization is slow, it may be convenient to dilute the lipase preparation at low ionic strength (even adding some glycerol).

3. Check that desorption of the lipase caused by organic solvents is not produced.

4. It is first necessary to check the effect of a similar amount of organic solvent during the assay in the enzyme activity; organic solvents may be inhibitors or activator of the lipases.

References

1. Undurraga, D., Markovits, A., and Erazo, S. (2001) Cocoa butter equivalent through enzymatic interesterification of palm oil midfraction. Process Biochem.

36,933-939.

2. Plou, J. F., Barandiaran, M., Calvo, M. V., Ballesteros, A., and Pastor, E. (1996) High yield production of mono and dioleylglycerol by lipase catalyzed hydrolysis of triolein. Enzyme Microb. Technol. 18, 66-71.

3. Jaeger, K.E. and Reetz, T.M. (1998) Microbial lipases from versatile tools for biotechnology. Trends Biotechnol. 16, 396-403.

4. Watanabe, Y., Miyawaki, Y., Adachi, S., Nakanishi, K., and Matsuno, R. (2001) Continuous production of acyl mannoses by immobilized lipase using a packed-bed reactor and their surfactant properties. Biochem. Eng. J. 8, 213-216.

5. Wong, C. H. and Whitesides, G. M. (1994) Enzymes in Synthetic Organic Chemistry, Tetrahedron Organic Chemistry Series, vol. 12, Pergamon Press, Oxford, UK.

6. Kazlauskas, R. J. and Bornscheuer, U. T. (1998) Biotransformations with Lipases in Biotechnology. 68-87.

7. Schmid, R.D. and Verger, R. (1998) Lipases: interfacial enzymes with attractive applications. Angew. Chem, Int. Ed. 37, 1609-1633.

8. Palomo, J. M., Fernandez-Lorente, G., Mateo, C., Fuentes, M., Fernandez-Lafuente, R., and Guisan, J. M. (2002) Modulation of the enantioselectivity of Candida antarctica B lipase via conformational engineering: kinetic resolution of (±)-a-hydroxy-phenylacetic acid derivatives. Tetrahedron: Asymmetry. 13, 13371345.

9. Palomo, J. M., Fernandez-Lorente, G., Rua, M. L., Guisan, J. M., and Fernandez-Lafuente, R. (2003) Evaluation of the lipase from Bacillus thermocatenulatus as an enantioselective biocatalyst. Tetrahedron: Asymmetry 14, 3679-3687.

10. Palomo, J. M., Mateo, C., Fernandez-Lorente, G., et al. (2003). Chiral resolution of (±)-5-substituted-6-(5-chloropyridin-2-yl)-7-oxo-5,6-dihydropyrrolo[3,4b]pyrazine derivatives-precursors of (S)-(+)-Zopiclone, catalyzed by immobilized C andida antarctica B lipase in aqueous media. Tetrahedron: Asymmetry 14, 429-438.

11. Brady, L., Brzozowski, A. M., Derewenda, Z. S., et al. (1990) A serine protease triad forms the catalytic center of a triacylglycerol lipase. Nature 343, 767-770.

12. Brzozowski, A. M., Derewenda, U., Derewenda, Z. S., et al. (1991) A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex. Nature 351, 491-494.

13. Sarda, L. and Desnuelle, P. (1958) Action de la lipase pancreatique sur les esteres en emulsion. Biochim. Biophys. Acta. 30, 513-521.

14. Ghosh, D., Wawrzak, Z., Pletnev, V. Z., et al. (1995) Structure of uncomplexed and linoleate-bound Candida cylindracea cholesterol esterase.Structure 3, 279-288.

15. García-Alles, L. F. and Gotor, V. (1998) Lipase-catalyzed transesterifcation in organic media: solvent effects on equilibrium and individual rate constants. Biotechnol. Bioeng. 59, 684-694.

16. Bastida, A., Sabuquillo, P., Armisen, P., Fernández-Lafuente, R., Huguet, J., and Guisán. J. M. (1998) A single step purification, immobilization and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol. Bioeng. 58, 486-493.

17. Palomo, J. M., Muñoz, G., Fernández-Lorente, G., Mateo, C., Fernández-Lafuente, R., and Guisán, J. M. (2002) Interfacial adsorption of lipases on very hydrophobic support (Octadecyl-Sepabeads): immobilization, hyperactivation and stabilization of the open form of lipases. J. Mol. Catal. B: Enzym. 19-20, 279-286.

18. Miled, N., Beisson, F., de Caro, J., de Caro, A., Arondel V., and Verger, R. (2001) Interfacial catalysis by lipases. J. Mol. Catal. B: Enzym. 11, 65-171.

19. Palomo, J. M., Segura, R. L., Fernández-Lorente, G., et al. (2004) Purification, Immobilization and stabilization of a lipase from Bacillus thermocatenulatus. Biotechnol. Prog. 20, 630-635.

20. Sabuquillo, P., Reina, J., Fernández-Lorente, G., Guisán, J. M., and Fernández-Lafuente, R. (1998) Interfacial affinity chromatography of lipases: separation of different fractions by selective adsorption on supports activated with hydrophobic groups.Biochim. Biophys. Acta. 1388, 337-348.

21. Guisán, J. M. (1988) Aldehyde gels as activated support for immobilization-stabilization of enzymes.Enzyme Microb. Technol. 10, 375-382.

22. Fernández-Lafuente, R., Rodríguez, V., and Guisán, J. M. (1998) The coimmobilization of D-aminoacid oxidase and catalase enables the quantitative transformation of D-amino acids (phenylalanine) into a-ceto acids (phenylpyru-vic acid). Enzyme Microb. Technol. 23, 28-33.

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