1vrx
From Proteopedia
OCA (Talk | contribs)
(New page: 200px<br /><applet load="1vrx" size="450" color="white" frame="true" align="right" spinBox="true" caption="1vrx, resolution 2.40Å" /> '''Endocellulase e1 fro...)
Next diff →
Revision as of 02:58, 21 November 2007
|
Endocellulase e1 from acidothermus cellulolyticus mutant y245g
Overview
<When Tyr245 in endocellulase Cel5A from Acidothermus cellulolyticus was, changed to Gly (Y245G) by designed mutation, the value of Ki for, inhibition of the enzyme by the product cellobiose was increased more than, 1480%. This reduction in product inhibition enabled the mutant enzyme, (used in conjunction with Trichoderma reesei cellobiohydrolase-I) to, release soluble sugars from biomass cellulose at a rate as much as 40%, greater than that achieved by the wild-type (WT) enzyme. The mutant was, designed on the basis of the previously published crystal structure of the, WT enzyme/substrate complex (at a resolution of 2.4 A), which provided, insights into the enzyme mechanism at the atomic level and identified, Tyr245 as a key residue interacting with a leaving group. To determine the, origin of the change in activity, the crystal structure of Y245G was, solved at 2.4-A resolution to an R-factor of 0.19 (R-free = 0.25). To, obtain additional information on the enzyme-product interactions, density, functional calculations were performed on representative fragments of the, WT Cel5A and Y245G. The combined results indicate that the loss of the, platform (Y245G) and of a hydrogen bond (from a conformational change in, Gln247) reduces the binding energy between product and enzyme by several, kilo calories per mole. Both kinetic and structural analyses thus relate, the increased enzymatic activity to reduced product inhibition.
About this Structure
1VRX is a Single protein structure of sequence from Acidothermus cellulolyticus. This structure superseeds the now removed PDB entry 1C0D. Active as Cellulase, with EC number 3.2.1.4 Full crystallographic information is available from OCA.
Reference
Catalytically enhanced endocellulase Cel5A from Acidothermus cellulolyticus., Baker JO, McCarley JR, Lovett R, Yu CH, Adney WS, Rignall TR, Vinzant TB, Decker SR, Sakon J, Himmel ME, Appl Biochem Biotechnol. 2005 Spring;121-124:129-48. PMID:15917594
Page seeded by OCA on Wed Nov 21 05:05:50 2007
