2F6C

Reaction geometry and thermostability of pyranose 2-oxidase from the white-rot fungus Peniophora sp., Thermostability mutant E542K

Summary for 2F6C

• Entry DOI: 10.2210/pdb2f6c/pdb
• Related: 1TZL 2F5V
• Descriptor: Pyranose 2-oxidase, FLAVIN-ADENINE DINUCLEOTIDE, TETRAETHYLENE GLYCOL, ... (5 entities in total)
• Functional Keywords: flavoprotein, covalent, histidine-bound, lignin degradation, thermostability mutation, d2 tetramer, phbh fold, gmc oxidoreductase, glutathione-reductase related fold, oxidoreductase
• Biological source: Peniophora sp. SG
• Cellular location: Periplasm : Q8J136
• Total number of polymer chains: 1
• Total formula weight: 67598.58

Authors

Bannwarth, M.,Heckmann-Pohl, D.M.,Bastian, S.,Giffhorn, F.,Schulz, G.E. (deposition date: 2005-11-29, release date: 2006-06-13, Last modification date: 2024-11-20)

Primary citation

Bannwarth, M.,Heckmann-Pohl, D.,Bastian, S.,Giffhorn, F.,Schulz, G.E.
Reaction Geometry and Thermostable Variant of Pyranose 2-Oxidase from the White-Rot Fungus Peniophora sp.
Biochemistry, 45:6587-6595, 2006

PubMed Abstract: Pyranose 2-oxidase catalyzes the oxidation of a number of carbohydrates using dioxygen; glucose, for example, is oxidized at carbon 2. The structure of pyranose 2-oxidase with the reaction product 2-keto-beta-d-glucose bound in the active center is reported in a new crystal form at 1.41 A resolution. The binding structure suggests that the alpha-anomer cannot be processed. The binding mode of the oxidized product was used to model other sugars accepted by the enzyme and to explain its specificity and catalytic rates. The reported structure at pH 6.0 shows a drastic conformational change in the loop of residues 454-461 (loop 454-461) at the active center compared to that of a closely homologous enzyme analyzed at pH 4.5 with a bound acetate inhibitor. In our structures, the loop is highly mobile and shifts to make way for the sugar to pass into the active center. Presumably, loop 454-461 functions as a gatekeeper. Apart from the wild-type enzyme, a thermostable variant was analyzed at 1.84 A resolution. In this variant, Glu542 is exchanged for a lysine. The observed stabilization could be a result of the mutated residue changing an ionic contact at a comparatively weak interface of the tetramer.

PubMed: 16716069
DOI: 10.1021/bi052465d

Experimental method

X-RAY DIFFRACTION (1.84 Å)