Gene cloning, heterologous overexpression and optimized refolding of the NAD-glutamate dehydrogenase from Haloferax mediterranei.
Extremophiles
; 10(2): 105-15, 2006 Apr.
Article
em En
| MEDLINE
| ID: mdl-16200391
ABSTRACT
The NAD-dependent glutamate dehydrogenase (GDH) gene from the halophilic archaeon Haloferax mediterranei has been cloned. The analysis of the nucleotide sequence revealed an open reading frame of 1323 bp that encodes a NAD-GDH. The amino acid sequence displayed high homology with those from other sources, especially the highly conserved residues involved in 2-oxoglutarate binding. The expression of this gene in Escherichia coli, the refolding and further characterization, yielded a fully active NAD-GDH with the same features than those found for the wild-type enzyme. This halophilic NAD-GDH showed a highly dependence on salts for both stability and activity, being essential for the refolding of the recombinant enzyme.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Haloferax mediterranei
/
Glutamato Desidrogenase
Idioma:
En
Revista:
Extremophiles
Assunto da revista:
BIOLOGIA
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
Espanha