Preliminary crystallographic studies of glucose dehydrogenase from the promiscuous Entner-Doudoroff pathway in the hyperthermophilic archaeon Sulfolobus solfataricus.
The hyperthermophilic archaeon Sulfolobus solfataricus grows optimally above 353 K and can metabolize glucose and its C4 epimer galactose via a non-phosphorylative variant of the Entner-Doudoroff pathway involving catalytically promiscuous enzymes that can operate with both sugars. The initial oxidation step is catalysed by glucose dehydrogenase (SsGDH), which can utilize both NAD and NADP as cofactors. The enzyme operates with glucose and galactose at similar catalytic efficiency, while its substrate profile also includes a range of other five- and six-carbon sugars. Crystals of the 164 kDa SsGDH homotetramer have been grown under a variety of conditions. The best crystals to date diffract to 1.8 A on a synchrotron source, have orthorhombic symmetry and belong to space group P2(1)2(1)2. Attempts are being made to solve the structure by MAD and MR.
Theodossis
Alex
A
Centre for Biomolecular Sciences, University of St Andrews, North Haugh, St Andrews, Fife KY16 9ST, Scotland.
Milburn
Christine C
CC
Heyer
Narinder I
NI
Lamble
Henry J
HJ
Hough
David W
DW
Danson
Michael J
MJ
Taylor
Garry L
GL
D019843:Archaeal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006358:Hot Temperature; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D048229:Sulfolobus solfataricus / Q000201:enzymology*; D017356:Synchrotrons; D013816:Thermodynamics; D014961:X-Ray Diffraction
D019843:Archaeal Proteins; D011994:Recombinant Proteins; D042843:Glucose 1-Dehydrogenase