spacer
spacer

PDBsum entry 2pwe

Go to PDB code: 
protein ligands metals Protein-protein interface(s) links
Isomerase PDB id
2pwe
Jmol
Contents
Protein chains
556 a.a. *
Ligands
SUC ×2
Metals
_CA ×2
Waters ×1501
* Residue conservation analysis
PDB id:
2pwe
Name: Isomerase
Title: Crystal structure of the mutb e254q mutant in complex with the substrate sucrose
Structure: Sucrose isomerase. Chain: a, b. Engineered: yes. Mutation: yes
Source: Pseudomonas mesoacidophila. Organism_taxid: 265293. Strain: mx-45. Gene: mutb. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
2.00Å     R-factor:   0.190     R-free:   0.235
Authors: S.Ravaud,X.Robert,R.Haser,N.Aghajari
Key ref:
S.Ravaud et al. (2007). Trehalulose synthase native and carbohydrate complexed structures provide insights into sucrose isomerization. J Biol Chem, 282, 28126-28136. PubMed id: 17597061 DOI: 10.1074/jbc.M704515200
Date:
11-May-07     Release date:   26-Jun-07    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
Q2PS28  (Q2PS28_9PSED) -  Sucrose isomerase (Precursor)
Seq:
Struc:
 
Seq:
Struc:
584 a.a.
556 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     carbohydrate metabolic process   1 term 
  Biochemical function     catalytic activity     4 terms  

 

 
DOI no: 10.1074/jbc.M704515200 J Biol Chem 282:28126-28136 (2007)
PubMed id: 17597061  
 
 
Trehalulose synthase native and carbohydrate complexed structures provide insights into sucrose isomerization.
S.Ravaud, X.Robert, H.Watzlawick, R.Haser, R.Mattes, N.Aghajari.
 
  ABSTRACT  
 
Various diseases related to the overconsumption of sugar make a growing need for sugar substitutes. Because sucrose is an inexpensive and readily available d-glucose donor, the industrial potential for enzymatic synthesis of the sucrose isomers trehalulose and/or isomaltulose from sucrose is large. The product specificity of sucrose isomerases that catalyze this reaction depends essentially on the possibility for tautomerization of sucrose, which is required for trehalulose formation. For optimal use of the enzyme, targeting controlled synthesis of these functional isomers, it is necessary to minimize the side reactions. This requires an extensive analysis of substrate binding modes and of the specificity-determining sites in the structure. The 1.6-2.2-A resolution three-dimensional structures of native and mutant complexes of a trehalulose synthase from Pseudomonas mesoacidophila MX-45 mimic successive states of the enzyme reaction. Combined with mutagenesis studies they give for the first time thorough insights into substrate recognition and processing and reaction specificities of these enzymes. Among the important outcomes of this study is the revelation of an aromatic clamp defined by Phe(256) and Phe(280) playing an essential role in substrate recognition and in controlling the reaction specificity, which is further supported by mutagenesis studies. Furthermore, this study highlights essential residues for binding the glucosyl and fructosyl moieties. The introduction of subtle changes informed by comparative three-dimensional structural data observed within our study can lead to fundamental modifications in the mode of action of sucrose isomerases and hence provide a template for industrial catalysts.
 
  Selected figure(s)  
 
Figure 1.
FIGURE 1. Schematic drawings of the products trehalulose (A) and isomaltulose (B) and of the substrate sucrose (C).
Figure 5.
FIGURE 5. Close up on the active sites of native MutB (A), E254Q·sucrose (B), MutB·castanospermine (C), MutB·deoxynojirimycin (D), and D200A·glucose (E). Electron density 2F[o] - F[c] maps are contoured at 1 . Catalytic residues are highlighted in green, and other active site residues are in yellow. The aromatic clamp in magenta corresponds to conformation 1, and that in cyan corresponds to conformation 2.
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 28126-28136) copyright 2007.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20589449 S.Li, H.Cai, Y.Qing, B.Ren, H.Xu, H.Zhu, and J.Yao (2011).
Cloning and characterization of a sucrose isomerase from Erwinia rhapontici NX-5 for isomaltulose hyperproduction.
  Appl Biochem Biotechnol, 163, 52-63.  
20875088 W.M.Patrick, Y.Nakatani, S.M.Cutfield, M.L.Sharpe, R.J.Ramsay, and J.F.Cutfield (2010).
Carbohydrate binding sites in Candida albicans exo-β-1,3-glucanase and the role of the Phe-Phe 'clamp' at the active site entrance.
  FEBS J, 277, 4549-4561.
PDB codes: 2pc8 2pf0 3n9k 3o6a
19966417 E.Champion, M.Remaud-Simeon, L.K.Skov, J.S.Kastrup, M.Gajhede, and O.Mirza (2009).
The apo structure of sucrose hydrolase from Xanthomonas campestris pv. campestris shows an open active-site groove.
  Acta Crystallogr D Biol Crystallogr, 65, 1309-1314.
PDB code: 2wpg
19783746 H.Watzlawick, and R.Mattes (2009).
Gene cloning, protein characterization, and alteration of product selectivity for the trehalulose hydrolase and trehalulose synthase from "Pseudomonas mesoacidophila" MX-45.
  Appl Environ Microbiol, 75, 7026-7036.  
19486422 J.Cha, J.H.Jung, S.E.Park, M.H.Cho, D.H.Seo, S.J.Ha, J.W.Yoon, O.H.Lee, Y.C.Kim, and C.S.Park (2009).
Molecular cloning and functional characterization of a sucrose isomerase (isomaltulose synthase) gene from Enterobacter sp. FMB-1.
  J Appl Microbiol, 107, 1119-1130.  
18494783 A.Godány, B.Vidová, and S.Janecek (2008).
The unique glycoside hydrolase family 77 amylomaltase from Borrelia burgdorferi with only catalytic triad conserved.
  FEMS Microbiol Lett, 284, 84-91.  
18398906 T.Shirai, V.S.Hung, K.Morinaka, T.Kobayashi, and S.Ito (2008).
Crystal structure of GH13 alpha-glucosidase GSJ from one of the deepest sea bacteria.
  Proteins, 73, 126-133.
PDB code: 2ze0
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. Where a reference describes a PDB structure, the PDB codes are shown on the right.