1w1k: Difference between revisions
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[[Image:1w1k.gif|left|200px]]<br /> | [[Image:1w1k.gif|left|200px]]<br /><applet load="1w1k" size="450" color="white" frame="true" align="right" spinBox="true" | ||
<applet load="1w1k" size="450" color="white" frame="true" align="right" spinBox="true" | |||
caption="1w1k, resolution 2.55Å" /> | caption="1w1k, resolution 2.55Å" /> | ||
'''STRUCTURE OF THE OCTAMERIC FLAVOENZYME VANILLYL-ALCOHOL OXIDASE: ILE238THR MUTANT'''<br /> | '''STRUCTURE OF THE OCTAMERIC FLAVOENZYME VANILLYL-ALCOHOL OXIDASE: ILE238THR MUTANT'''<br /> | ||
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==About this Structure== | ==About this Structure== | ||
1W1K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with FAD and EUG as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alcohol_oxidase Alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.13 1.1.3.13] | 1W1K is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with FAD and EUG as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alcohol_oxidase Alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.13 1.1.3.13] Known structural/functional Site: <scene name='pdbsite=AC1:Eug Binding Site For Chain B'>AC1</scene>. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1W1K OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: oxidoreductase]] | [[Category: oxidoreductase]] | ||
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Dec 18 18:26:15 2007'' | ||
Revision as of 19:16, 18 December 2007
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STRUCTURE OF THE OCTAMERIC FLAVOENZYME VANILLYL-ALCOHOL OXIDASE: ILE238THR MUTANT
OverviewOverview
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.
About this StructureAbout this Structure
1W1K is a Single protein structure of sequence from Penicillium simplicissimum with FAD and EUG as ligands. Active as Alcohol oxidase, with EC number 1.1.3.13 Known structural/functional Site: . Full crystallographic information is available from OCA.
ReferenceReference
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
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