RT Journal Article
T1 Immobilization–stabilization of a new recombinant glutamate dehydrogenase from Thermus thermophilus
A1 Bolívar Bolívar, Juan Manuel
A1 Cava, Felipe
A1 Mateo, Cesar
A1 Rocha Martín, Javier
A1 Guisán, Jose M.
A1 Berenguer, José
A1 Fernandez-Lafuente, Roberto
AB The genome of Thermus thermophilus contains two genes encoding putative glutamate dehydrogenases. One of these genes (TTC1211) was cloned and overexpressed in Escherichia coli. The purified enzyme was a trimer that catalyzed the oxidation of glutamate to α-ketoglutarate and ammonia with either NAD+ or NADP+ as cofactors. The enzyme was also able to catalyze the inverse reductive reaction. The thermostability of the enzyme at neutral pH was very high even at 70°C, but at acidic pH values, the dissociation of enzyme subunits produced the rapid enzyme inactivation even at 25°C. The immobilization of the enzyme on glyoxyl agarose permitted to greatly increase the enzyme stability under all conditions studied. It was found that the multimeric structure of the enzyme was stabilized by the immobilization (enzyme subunits could be not desorbed from the support by boiling it in the presence of sodium dodecyl sulfate). This makes the enzyme very stable at pH 4 (e.g., the enzyme activity did not decrease after 12 h at 45°C) and even improved the enzyme stability at neutral pH values. This immobilized enzyme can be of great interest as a biosensor or as a biocatalyst to regenerate both reduced and oxidized cofactors.
PB Springer
SN 0175-7598
YR 2008
FD 2008
LK https://hdl.handle.net/20.500.14352/92757
UL https://hdl.handle.net/20.500.14352/92757
LA eng
NO Bolivar, J. M., Cava, F., Mateo, C., Rocha-Martín, J., Guisán, J. M., Berenguer, J., & Fernandez-Lafuente, R. (2008). Immobilization-stabilization of a new recombinant glutamate dehydrogenase from Thermus thermophilus. Applied Microbiology and Biotechnology, 80(1), 49-58. https://doi.org/10.1007/S00253-008-1521-3
NO Comunidad de Madrid
NO Ministerio de Educación y Ciencia (España)
NO Fundación Ramón Areces (España)
DS Docta Complutense
RD 7 abr 2025