O. Barbosa et al. / Journal of Molecular Catalysis B: Enzymatic 71 (2011) 124–132
131
mixture, while for all the other preparations the decrement on the
E values was much lower.
Thus, the change of the solvent presented very different effects
on the activities and enantiospecificities of CalB immobilized fol-
lowing different immobilization protocols, perhaps by modulating
the conformational changes induced by the different solvents.
[11] R. Fernandez-Lafuente, P. Armisén, P. Sabuquillo, G. Fernández-Lorente, J.M.
Guisan, Chem. Phys. Lipids 93 (1998) 185–197.
[
[
12] M.T. Reetz, Tetrahedron 58 (2002) 6595–6602.
13] D. Rotticci, J. Rotticci-Mulder, S. Denman, T. Norin, K. Hult, Chem. Biochem. 2
(2001) 766–770.
[
14] F. Secundo, G. Carrea, C. Tarabiono, P. Lafranconi, S. Brocca, M.M. Lotti, et al., J.
Mol. Catal. B: Enzym. 39 (2006) 166–170.
[
[
15] A. Svendsen, Biochem. Biophys. Acta 1543 (2000) 223–238.
16] P. Villeneuve, J. Muderhwa, J. Graille, M. Haas, J. Mol. Catal. B Enzym. 9 (2000)
1
13–148.
4
. Conclusions
[
[
17] R. Patel, Coordin. Chem. Rev. 252 (2008) 659–701.
18] D. Zelaszczyk, K. Kiec-Kononowicz, Curr. Med. Chem. 14 (2007) 53–65.
This paper describes the preparation of different CalB biocata-
[19] D. Subhas, A. Venkat, Bioorg. Med. Chem. 13 (2005) 627–630.
[
[
20] H.S. Bevinakatti, A.A. Banerji, J. Org. Chem. 57 (1992) 6003–6005.
21] B.D. Brandes, E.N. Jacobsen, Tetrahedron Asymmetry 23 (1997) 3927–
lysts based on the epoxy covalent attachment but with alterations
in the first step of enzyme immobilization by hydrophobic, ionic
exchange and immobilized metal chelates adsorption for producing
biocatalysts with the same enzyme and covalent immobilization
chemistry but with a different orientation of the enzyme on the
support surface [61].
The immobilized preparations of CalB exhibited very different
kinetic properties, giving very different maximum synthetic yields
and enantiospecificities, although in all cases the activity lost per
reaction cycle did not exceed from 12%. The best preparation, EC-
CalB, reached a 76% maximum yield with an E value of 65, while the
less suitable preparation gave a maximum yield of only 27% and an
E value of 6. EC-CalB was used four times with a total decrease in
the enzyme activity of 40% that was possibly due to acetylation of
the enzyme by vinyl acetate.
The modulation of CalB by immobilization had been shown in
many instances in aqueous medium (enantiospecific hydrolysis in
resolution of racemic mixtures [65], regioselective hydrolysis of
sugars [66], specific and regioselective hydrolysis on production
of 1,2 diacetin [67], etc.). In this paper, the use of different immo-
bilization protocols has been shows as a powerful tool to alter
lipase properties, also in organic medium. Further optimization of
the reaction conditions and assay of new biocatalysts with a more
controlled immobilization [61] may permit to further increase the
performance of CalB on this reaction. Though the study presented
in this paper is specific to one drug, the outcome may be used for
other important chiral intermediates and chemicals.
3
933.
[22] A.O. Magnusson, J.C. Rotticci-Mulder, A. Santagostino, K. Hult, ChemBioChem
(2005) 1051–1056.
23] A. Magnusson, M. Takwa, M.A. Hamberg, K. Hult, Angew. Chem. Int. Ed. 44
2005) 4582–4585.
6
[
(
[24] Z. Marton, V. Léonard-Nevers, P. Syrén, C. Bauer, S. Lamare, K.K. Hult, et al., J.
Mol. Catal. B: Enzym. 65 (2010) 11–17.
[
25] C. Mateo, J.M. Palomo, G. Fernandez-Lorente, J.M. Guisan, R. Fernández-
Lafuente, Enzyme Microb. Technol. 40 (2007) 1451–1463.
[26] J. Kazlauskas, A. Weissfloch, A. Rappaport, L.A. Cuccia, J. Org. Chem. 56 (1991)
656–2665.
27] D. Rotticci, F. Haeffner, C. Orrenius, T. Norin, K. Hult, J. Mol. Catal. B: Enzym. 5
1998) 267–272.
[28] B. Hwang, H. Scheib, J. Pleiss, B. Kim, R. Schmid, J. Mol. Catal. B: Enzym. 10
2000) 223–231.
29] J. Ottosson, L. Fransson, J. King, K. Hult, Biochem. Biophys. Acta 1594 (2002)
25–334.
2
[
(
(
[
3
[30] A. Ducret, M. Trani, R. Lortie, Enzyme Microb. Technol. 22 (1998) 212–
216.
[
[
[
[
31] E. Wehtje, D. Costes, P. Adlercreutz, J. Mol. Catal. B: Enzym. 3 (1997) 221–
30.
32] M. Persson, D. Costes, E. Wehtje, P. Adlercreutz, Enzyme Microb. Technol. 30
(2002) 916–923.
2
33] A. Jönsson, W. Wehtje, P. Adlercreutz, B. Mattiasson, Biochem. Biophys. Acta
1430 (1999) 313–322.
34] R. Phillips, Enzyme Microb. Technol. 14 (1992) 417–419.
[35] R. Phillips, Trends Biotechnol. 14 (1996) 13–16.
[
[
[
36] E. Anderson, K. Larsson, O. Kirk, Biocatal. Biotransform. 16 (1998) 181–204.
37] J. Uppenberg, M. Hansen, S. Patkar, T. Jones, Structure 2 (1994) 293–308.
38] J. Uppenberg, N. Ohrner, M. Norin, K. Hult, G. Kleywegt, S. Patkar, et al., Bio-
chemistry 34 (1995) 16838–16851.
[
[
39] V. Gotor-Fernández, E. Busto, V. Gotor, Adv. Synth. Catal. 348 (2006) 797–812.
40] Y. Shimada, Y. Watanabe, A. Sugihara, Y. Tominaga, J. Mol. Catal. B Enzym. 17
(
2002) 133–142.
[
[
41] O. Kirk, M. Wurtz, Org. Process Res. Dev. 6 (2002) 446–451.
42] M. Skjøt, L. de Maria, R. Chatterjee, A. Svendsen, S.A. Patkar, P.R. Østergaard, J.
Brask, ChemBioChem 10 (2009) 520–527.
Acknowledgements
[
[
43] E. Katchalski-Katzir, D. Kraemer, J. Mol. Catal. B: Enzym. 10 (2000) 157–176.
44] C. Mateo, V. Grazú, B.C.C. Pessela, T. Montes, T.J.M. Palomo, R. Torres, et al.,
Biochem. Soc. Trans. 35 (2007) 1593–1601.
45] J. Turkova, K. Blaha, M. Malanikova, Biochem. Biophys. Acta 524 (1978)
162–169.
This work was supported by Grant No. 1102-489-25428 from
COLCIENCIAS and Universidad Industrial de Santander (VIE-UIS
Research Program) and grant CTQ2009-07568 from Spanish Min-
isterio de Ciencia e Innovación. The support of COLCIENCIAS (PhD
Program Fellowoship-2008) is gratefully recognized. Authors also
gratefully recognize the kind supply of Eupergit C by Rohm Pharma
[
[46] J.B. Wheatley, D.E. Schmidt Jr., J. Chromatogr. A 849 (1999) 1–12.
[
47] J.B. Wheatley, M.H. Lyttle, M.D. Hocker, D.E. Schmidt Jr., J. Chromatogr. A 726
1996) 77–90.
48] J.B. Wheatley, D.E. Schmidt Jr., J. Chromatogr. 644 (1993) 11–16.
(
[
(
Darstamdt, Germany). The help and comments from Dr. Ángel
[49] C. Mateo, G. Fernández-Lorente, O. Abian, R. Fernández-Lafuente, J.M. Guisán,
Biomacromolecules 1 (2000) 739–745.
Berenguer (Instituto de Materiales, Universidad de Alicante) and
Robert Murdoch (Institute for Comparative and Environmental
Toxicology, Cornell University) are gratefully recognized.
[
50] A. Bastida, P. Sabuquillo, P. Armisén, R. Fernández-Lafuente, J. Huguet, J.M.
Guisan, Biotechnol. Bioeng. 58 (1998) 486–493.
[51] G. Fernandez-Lorente, Z. Cabrera, Z.C. Godoy, R. Fernandez-Lafuente, J.M.
Palomo, J.M. Guisan, Process Biochem. 43 (2008) 1061–1067.
[
[
[
[
[
52] P. Sabuquillo, J. Reina, G. Fernández-Lorente, J.M. Guisán, R. Fernández-
Lafuente, Biochem. Biophys. Acta 1388 (1998) 337–348.
53] C. Mateo, R. Torres, G. Fernández-Lorente, C. Ortiz, C.M. Fuentes, A. Hidalgo,
et al., Biomacromolecules 4 (2003) 772–777.
54] C. Mateo, O. Abian, R. Fernández-Lafuente, J.M. Guisán, Enzyme Microb. Tech-
nol. 26 (2000) 509–515.
55] C. Mateo, O. Abian, G. Fernández-Lorente, J. Pedroche, R. Fernández-Lafuente,
J.M. Guisan, et al., Biotechnol. Prog. 18 (2002) 629–634.
56] J.E. Oh, K.-W. Lee, H.-K. Park, J.-Y. Kim, K.I.L. Kwon, J.-W. Kim, et al., J. Agric.
Food Chem. 57 (2009) 9280–9283.
57] R. Valivety, P. Halling, A. Macrae, Biochem. Biophys. Acta 1118 (1992) 218–222.
58] C.-S. Chen, S.-H. Wu, G. Girdaukas, C. Sih, J. Am. Chem. Soc. 109 (1987)
References
[
[
[
1] A. Ghanem, H. Aboul-Enein, Tetrahedron Asymmetry 15 (2004) 3331–3351.
2] A. Ghanem, Tetrahedron 63 (2007) 1721–1754.
3] V. Gotor-Fernández, R. Brieva, V. Gotor, J. Mol. Catal. B: Enzym. 40 (2006)
1
11–120.
4] M. Noble, A. Cleasby, L. Johnson, M. Egmond, L. Frenken, FEBS Lett. 331 (1993)
23–128.
[
1
[
[
5] Z.S. Derewenda, U. Derewenda, G.G. Dodson, J. Mol. Biol. 227 (1992) 818–839.
6] C. Carrasco-López, C. Godoy, B. de las Rivas, G. Fernández-Lorente, J.M. Palomo,
J.M. Guisán, et al., J. Biol. Chem. 284 (2009) 4365–4372.
[
[
2812–2817.
[7] H. Van Tilbeurgh, P. Egloff, C. Martinez, N. Rugani, R. Verger, C. Cambillau, Nature
[
[
[
59] V. Kasche, Enzyme Microb. Technol. 8 (1986) 4–16.
3
62 (1993) 814–820.
60] V. Kasche, U. Haufler, L. Riechmann, Method Enzym. 136 (1987) 280–292.
61] K. Hernandez, R. Fernandez-Lafuente, Enzyme Microb. Technol. (2011),
doi:10.1016/j.enzmictec.2010.10.003.
62] M. Fuentes, P. Batalla, V. Grazu, B.C.C. Pessela, C. Mateo, T.T. Montes, et al.,
Biomacromolecules 8 (2007) 703–707.
[
[
8] R. Verger, Trends Biotechnol. 15 (1997) 32–38.
9] J.M. Palomo, M. Pe n˜ as, G. Fernández-Lorente, C. Mateo, C.A. Pisabarro, R.
Fernández-Lafuente, et al., Biomacromolecules 4 (2003) 204–210.
[
[10] J.M. Palomo, M. Fuentes, M.G. Fernández-Lorente, C. Mateo, J.M. Guisán, R.
Fernández-Lafuente, Biomacromolecules 4 (2003) 1–6.