Characterization of a thermostable glucose dehydrogenase with strict substrate specificity from a hyperthermophilic archaeon Thermoproteus sp. GDH-1.
A hyperthermophilic archaeon was isolated from a terrestrial hot spring on Kodakara Island, Japan and designated as Thermoproteus sp. glucose dehydrogenase (GDH-1). Cell extracts from cells grown in medium supplemented with glucose exhibited NAD(P)-dependent glucose dehydrogenase activity. The enzyme (TgGDH) was purified and found to display a strict preference for D-glucose. The gene was cloned and expressed in Escherichia coli, resulting in the production of a soluble and active protein. Recombinant TgGDH displayed extremely high thermostability and an optimal temperature higher than 85 °C, in addition to its strict specificity for D-glucose. Despite its thermophilic nature, TgGDH still exhibited activity at 25 °C. We confirmed that the enzyme could be applied for glucose measurements at ambient temperatures, suggesting a potential of the enzyme for use in measurements in blood samples.
a Institute of Biotechnology , TOYOBO CO., LTD. , Tsuruga , Japan.
Hiroshi
H
Aiba
Yoshiaki
Y
Nishiya
Masayuki
M
Azuma
Yuusuke
Y
Yokooji
Haruyuki
H
Atomi
Tadayuki
T
Imanaka
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007564:Japan; D007700:Kinetics; D008969:Molecular Sequence Data; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D013379:Substrate Specificity; D013696:Temperature; D041542:Thermoproteus / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification
D019843:Archaeal Proteins; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D042843:Glucose 1-Dehydrogenase; D005947:Glucose
archaea; glucose dehydrogenase (GDH); hyperthermophile; thermostability