1w1m | pdb_00001w1m

STRUCTURE OF THE OCTAMERIC FLAVOENZYME VANILLYL-ALCOHOL OXIDASE: GLU502GLY MUTANT

spinqualitylabelsall models 1w1m, resolution 3.00Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 1w1m is a 2 chain structure with sequence from Penicillium simplicissimum. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: EUG, FAD Related: 1ahu, 1ahv, 1ahz, 1dzn, 1e0y, 1e8f, 1e8g, 1e8h, 1qlt, 1qlu, 1vao, 2vao, 1w1j, 1w1l, 1w1k Activity: Alcohol oxidase, with EC number 1.1.3.13 Resources: FirstGlance, OCA, RCSB, PDBsum Evolutionary Conservation Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf. Publication Abstract from PubMed The flavoenzyme vanillyl-alcohol oxidase was subjected to random mutagenesis to generate mutants with enhanced reactivity to creosol (2-methoxy-4-methylphenol). The vanillyl-alcohol oxidase-mediated conversion of creosol proceeds via a two-step process in which the initially formed vanillyl alcohol (4-hydroxy-3-methoxybenzyl alcohol) is oxidized to the widely used flavor compound vanillin (4-hydroxy-3-methoxybenzaldehyde). The first step of this reaction is extremely slow due to the formation of a covalent FAD N-5-creosol adduct. After a single round of error-prone PCR, seven mutants were generated with increased reactivity to creosol. The single-point mutants I238T, F454Y, E502G, and T505S showed an up to 40-fold increase in catalytic efficiency (kcat/Km) with creosol compared with the wild-type enzyme. This enhanced reactivity was due to a lower stability of the covalent flavin-substrate adduct, thereby promoting vanillin formation. The catalytic efficiencies of the mutants were also enhanced for other ortho-substituted 4-methylphenols, but not for p-cresol (4-methylphenol). The replaced amino acid residues are not located within a distance of direct interaction with the substrate, and the determined three-dimensional structures of the mutant enzymes are highly similar to that of the wild-type enzyme. These results clearly show the importance of remote residues, not readily predicted by rational design, for the substrate specificity of enzymes. Laboratory-evolved vanillyl-alcohol oxidase produces natural vanillin.,van den Heuvel RH, van den Berg WA, Rovida S, van Berkel WJ J Biol Chem. 2004 Aug 6;279(32):33492-500. Epub 2004 May 28. PMID:15169773 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Vanillyl-alcohol oxidase

Proteopedia Page Contributors and Editors (what is this?)

OCA