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

• Published in: Journal of molecular biology Vol. 223; no. 4; pp. 1167 - 1169
• Main Authors: Hendle, Jörg, Hecht, Hans-Jürgen, Kalisz, Henryk M., Schmid, Rolf D., Schomburg, Dietmar
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 20.02.1992; Elsevier
• Subjects: Biological and medical sciences; crystal structure; Crystalline structure; crystallization; Crystallography; crystals; dimensions; enzymology; FAD; Fundamental and applied biological sciences. Psychology; glucose oxidase; Glucose Oxidase - ultrastructure; interaction; Molecular biophysics; Penicillium; Penicillium - enzymology; Penicillium amagasakiense; Protein Conformation; purification; Structure in molecular biology; ultrastructure; X-ray crystallography; X-Ray Diffraction; crystallization; Penicillium amagasakiense; glucose oxidase; Fungi; Glucose oxidase; Unit cell; Purification; Enzyme; Crystallization; Coenzyme enzyme complex; Fungi Imperfecti; X ray diffraction; Thallophyta
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/0022-2836(92)90267-N

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 Å resolution and belong to the orthorhombic space group P2 12 12 1, with refined lattice constants of a = 59.3 A ̊ , b = 136.3 A ̊ and c = 156.7 A ̊ . Assuming two monomers (≈ 135 kDa) per asymmetric unit the V m value is 2.3 Å 3/Da.