1t2x

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{{STRUCTURE_1t2x| PDB=1t2x | SCENE= }}
{{STRUCTURE_1t2x| PDB=1t2x | SCENE= }}
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'''Glactose oxidase C383S mutant identified by directed evolution'''
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===Glactose oxidase C383S mutant identified by directed evolution===
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==Overview==
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Galactose oxidase (GO; E.C. 1.1.3.9) is a copper- containing enzyme that oxidizes a range of primary alcohols to aldehydes. This broad substrate specificity is reflected in a high K(M) for substrates. Directed evolution has previously been used to select variants of GO that exhibit enhanced expression and kinetic properties. In assays using unpurified enzyme samples, the variant C383S displayed a 5-fold lower K(M) than wild-type GO. In the present study, we have constructed, expressed, purified and characterized a number of single, double and triple mutants at residues Cys383, Tyr436 and Val494, identified in one of the directed evolution studies, to examine their relative contributions to improved catalytic activity of GO. We report kinetic studies on the various mutant enzymes. In addition, we have determined the three-dimensional structure of the C383S variant. As with many mutations identified in directed evolution experiments, the availability of structural information does not provide a definitive answer to the reason for the improved K(M) in the C383S variant protein.
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(as it appears on PubMed at http://www.pubmed.gov), where 15047910 is the PubMed ID number.
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{{ABSTRACT_PUBMED_15047910}}
==About this Structure==
==About this Structure==
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[[Category: C383s mutant form]]
[[Category: C383s mutant form]]
[[Category: Mutant form of copper containing enzyme]]
[[Category: Mutant form of copper containing enzyme]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 09:26:58 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Jul 27 17:13:26 2008''

Revision as of 14:13, 27 July 2008

Image:1t2x.png

Template:STRUCTURE 1t2x

Glactose oxidase C383S mutant identified by directed evolution

Template:ABSTRACT PUBMED 15047910

About this Structure

Full crystallographic information is available from OCA.

Reference

Structural and kinetic studies of a series of mutants of galactose oxidase identified by directed evolution., Wilkinson D, Akumanyi N, Hurtado-Guerrero R, Dawkes H, Knowles PF, Phillips SE, McPherson MJ, Protein Eng Des Sel. 2004 Feb;17(2):141-8. Epub 2004 Jan 12. PMID:15047910

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