Heterologous overexpression of glucose dehydrogenase from the halophilic archaeon Haloferax mediterranei, an enzyme of the medium chain dehydrogenase/reductase family.
The first gene encoding a glucose dehydrogenase (GDH) from a halophilic organism has been sequenced. Amino acid sequence alignments of GDH from Haloferax mediterranei show a high degree of homology with the thermoacidophilic GDHs and with other enzymes from the medium chain dehydrogenase/reductase family. Heterologous overexpression using the mesophilic organism Escherichia coli as the host has been performed and the expression product was obtained as inclusion bodies. To obtain the halophilic enzyme in its native form refolding and reactivation in a saline environment were required. A pure and highly concentrated sample of the enzyme was obtained using a purification procedure based on the protein's halophilicity. This method may be useful as a general procedure for purifying other halophilic proteins from mesophilic hosts.
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División de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Alicante, Ap. 99, E-03080, Alicante, Spain.
Esclapez
J
J
Ferrer
J
J
Bonete
M J
MJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D019641:DNA, Archaeal / Q000032:analysis; D004926:Escherichia coli; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D019623:Haloferax mediterranei / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012995:Solubility
D019641:DNA, Archaeal; D011994:Recombinant Proteins; D010088:Oxidoreductases; D005948:Glucose Dehydrogenases; D042843:Glucose 1-Dehydrogenase