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PDBsum entry 2vuj

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Hydrolase PDB id
2vuj
Jmol
Contents
Protein chain
196 a.a. *
Ligands
GOL
Waters ×247
* Residue conservation analysis
PDB id:
2vuj
Name: Hydrolase
Title: Environmentally isolated gh11 xylanase
Structure: Gh11 xylanase. Chain: a. Engineered: yes
Source: Escherichia coli. Organism_taxid: 562. Expressed in: escherichia coli. Expression_system_taxid: 511693.
Resolution:
1.80Å     R-factor:   0.188     R-free:   0.241
Authors: C.Dumon,A.Varvak,M.A.Wall,J.E.Flint,R.J.Lewis,J.H.Lakey, P.Luginbuhl,S.Healey,T.Todaro,G.Desantis,M.Sun, L.Parra-Gessert,X.Tan,D.P.Weiner,H.J.Gilbert
Key ref:
C.Dumon et al. (2008). Engineering Hyperthermostability into a GH11 Xylanase Is Mediated by Subtle Changes to Protein Structure. J Biol Chem, 283, 22557-22564. PubMed id: 18515360 DOI: 10.1074/jbc.M800936200
Date:
26-May-08     Release date:   17-Jun-08    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
No UniProt id for this chain
Struc: 196 a.a.
Key:    Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.3.2.1.8  - Endo-1,4-beta-xylanase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Endohydrolysis of 1,4-beta-D-xylosidic linkages in xylans.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     carbohydrate metabolic process   1 term 
  Biochemical function     hydrolase activity, hydrolyzing O-glycosyl compounds     1 term  

 

 
DOI no: 10.1074/jbc.M800936200 J Biol Chem 283:22557-22564 (2008)
PubMed id: 18515360  
 
 
Engineering Hyperthermostability into a GH11 Xylanase Is Mediated by Subtle Changes to Protein Structure.
C.Dumon, A.Varvak, M.A.Wall, J.E.Flint, R.J.Lewis, J.H.Lakey, C.Morland, P.Luginbühl, S.Healey, T.Todaro, G.Desantis, M.Sun, L.Parra-Gessert, X.Tan, D.P.Weiner, H.J.Gilbert.
 
  ABSTRACT  
 
Understanding the structural basis for protein thermostability is of considerable biological and biotechnological importance as exemplified by the industrial use of xylanases at elevated temperatures in the paper pulp and animal feed sectors. Here we have used directed protein evolution to generate hyperthermostable variants of a thermophilic GH11 xylanase, EvXyn11. The Gene Site Saturation Mutagenesistrade mark (GSSM) methodology employed assesses the influence on thermostability of all possible amino acid substitutions at each position in the primary structure of the target protein. The 15 most thermostable mutants, which generally clustered in the N-terminal region of the enzyme, had melting temperatures (T(m)) 1-8 degrees C higher than the parent protein. Screening of a combinatorial library of the single mutants identified a hyperthermostable variant, EvXyn11(TS), containing seven mutations. EvXyn11(TS) had a T(m) approximately 25 degrees C higher than the parent enzyme while displaying catalytic properties that were similar to EvXyn11. The crystal structures of EvXyn11 and EvXyn11(TS) revealed an absence of substantial changes to identifiable intramolecular interactions. The only explicable mutations are T13F, which increases hydrophobic interactions, and S9P that apparently locks the conformation of a surface loop. This report shows that the molecular basis for the increased thermostability is extraordinarily subtle and points to the requirement for new tools to interrogate protein folding at non-ambient temperatures.
 
  Selected figure(s)  
 
Figure 1.
FIGURE 1. Graph showing how increasing the number of colonies screened increases the percentage coverage of the GSSM library.
Figure 6.
FIGURE 6. Relationship between tolerance to mutation with respect to activity at positions where amino acid substitutions confer increased thermostability. The x axis shows the proportion of substitutions at each position in the sequence (value of 1 signifies that the enzyme retains activity irrespective of the amino acid substitution at the specified position) that can be tolerated.
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2008, 283, 22557-22564) copyright 2008.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19458918 Y.M.Park, and S.Y.Ghim (2009).
Enhancement of the activity and pH-performance of chitosanase from Bacillus cereus strains by DNA shuffling.
  Biotechnol Lett, 31, 1463-1467.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.