Crystallization and preliminary X-ray diffraction studies of a deglycosylated glucose oxidase from Penicillium amagasakiense.
The dimeric glucose oxidase from Penicillium amagasakiense was deglycosylated, purified and crystallized as a complex with its coenzyme FAD. Deglycosylation and purification to isoelectric homogeneity were shown to be an important prerequisite step to obtain crystals suitable for X-ray investigations. Crystals of the deglycosylated enzyme were reproducibly grown using ammonium sulfate as precipitant at pH 7.4 to 7.5. Crystals diffract to at least 2.0 A resolution and belong to the orthorhombic space group P2(1)2(1)2(1), with refined lattice constants of a = 59.3 A, b = 136.3 A and c = 156.7 A. Assuming two monomers (approximately 135 kDa) per asymmetric unit the Vm value is 2.3 A3/Da.
Hendle
J
J
Department of Enzyme Technology, GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
Hecht
H J
HJ
Kalisz
H M
HM
Schmid
R D
RD
Schomburg
D
D
D003461:Crystallography; D005949:Glucose Oxidase / Q000648:ultrastructure*; D010407:Penicillium / Q000201:enzymology*; D011487:Protein Conformation; D014961:X-Ray Diffraction
D005949:Glucose Oxidase