Expression, purification and crystallization of a thermostable short-chain alcohol dehydrogenase from the archaeon Thermococcus sibiricus.
Alcohol dehydrogenases belong to the oxidoreductase family and play an important role in a broad range of physiological processes. They catalyze the cofactor-dependent reversible oxidation of alcohols to the corresponding aldehydes or ketones. The NADP-dependent short-chain alcohol dehydrogenase TsAdh319 from the thermophilic archaeon Thermococcus sibiricus was overexpressed, purified and crystallized. Crystals were obtained using the hanging-drop vapour-diffusion method using 25%(w/v) polyethylene glycol 3350 pH 7.5 as precipitant. The crystals diffracted to 1.68 A resolution and belonged to space group I222, with unit-cell parameters a = 55.63, b = 83.25, c = 120.75 A.
Lyashenko
A V
AV
Shubnikov Institute of Crystallography, RAS, Leninsky Prospekt 59, Moscow 119333, Russia. lyashenko@ns.crys.ras.ru
Bezsudnova
E Y
EY
Gumerov
V M
VM
Lashkov
A A
AA
Mardanov
A V
AV
Mikhailov
A M
AM
Polyakov
K M
KM
Popov
V O
VO
Ravin
N V
NV
Skryabin
K G
KG
Zabolotniy
V K
VK
Stekhanova
T N
TN
Kovalchuk
M V
MV
D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D015870:Gene Expression; D013696:Temperature; D019712:Thermococcus / Q000201:enzymology*
D000426:Alcohol Dehydrogenase