Crystallization and preliminary X-ray diffraction studies of a deglycosylated glucose oxidase from Penicillium amagasakiense

J Hendle; H J Hecht; H M Kalisz; R D Schmid; D Schomburg

doi:10.1016/0022-2836(92)90267-n

Crystallization and preliminary X-ray diffraction studies of a deglycosylated glucose oxidase from Penicillium amagasakiense

J Mol Biol. 1992 Feb 20;223(4):1167-9. doi: 10.1016/0022-2836(92)90267-n.

Authors

J Hendle¹, H J Hecht, H M Kalisz, R D Schmid, D Schomburg

Affiliation

¹ Department of Enzyme Technology, GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.

PMID: 1538394
DOI: 10.1016/0022-2836(92)90267-n

Abstract

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.

MeSH terms

Crystallography
Glucose Oxidase / ultrastructure*
Penicillium / enzymology*
Protein Conformation
X-Ray Diffraction

Substances

Glucose Oxidase