RT Journal Article
T1 Improvement of the stability of alcohol dehydrogenase by covalent immobilization on glyoxyl-agarose
A1 Bolívar Bolívar, Juan Manuel
A1 Wilson, Lorena
A1 Ferrarotti, Susana Alicia
A1 Guisán, José M.
A1 Fernández-Lafuente, Roberto
A1 Mateo, Cesar
AB Immobilization of alcohol dehydrogenase (ADH) from Horse Liver inside porous supports promotes a dramatic stabilization of the enzyme against inactivation by air bubbles in stirred tank reactors. Moreover, immobilization of ADH on glyoxyl-agarose promotes additional stabilization against any distorting agent (pH, temperature, organic solvents, etc.). Stabilization is higher when using highly activated supports, they are able to immobilize both subunits of the enzyme. The best glyoxyl derivatives are much more stable than conventional ADH derivatives (e.g., immobilized on BrCN activated agarose). For example, glyoxyl immobilized ADH preserved full activity after incubation at pH 5.0 for 20 h at room temperature and conventional derivatives (as well as the soluble enzyme) preserved less than 50% of activity after incubation under the same conditions. Moreover, glyoxyl derivatives are more than 10 times more stable than BrCN derivatives when incubated in 50% acetone at pH 7.0. Multipoint covalent immobilization, in addition to multisubunit immobilization, seems to play an important stabilizing role against distorting agents. In spite of these interesting stabilization factors, immobilization hardly promotes losses of catalytic activity (keeping values near to 90%). This immobilized preparation is able to keep good activity using dextran-NAD+. In this way, ADH glyoxyl immobilized preparation seems to be suitable to be used as cofactor-recycling enzyme-system in interesting NAD+-mediated oxidation processes, catalyzed by other immobilized dehydrogenases in stirred tank reactors
PB Elsevier
SN 0168-1656
YR 2006
FD 2006
LK https://hdl.handle.net/20.500.14352/93046
UL https://hdl.handle.net/20.500.14352/93046
LA eng
NO Bolivar, J. M., Wilson, L., Ferrarotti, S. A., Guisán, J. M., Fernández-Lafuente, R., & Mateo, C. (2006). Improvement of the stability of alcohol dehydrogenase by covalent immobilization on glyoxyl-agarose. Journal of Biotechnology, 125(1), 85-94. https://doi.org/10.1016/J.JBIOTEC.2006.01.028
NO Comunidad de Madrid
NO Ministerio de Educación y Ciencia (España)
NO Fundación Carolina (España)
NO Ministerio de Educación (Chile)
DS Docta Complutense
RD 14 jun 2025