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PDBsum entry 2wne
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Contents |
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* Residue conservation analysis
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PDB id:
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Hydrolase
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Title:
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Mutant laminarinase 16a cyclizes laminariheptaose
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Structure:
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Putative laminarinase. Chain: a. Fragment: residues 21-318. Synonym: laminarinase 16a. Engineered: yes. Mutation: yes. Other_details: nucleophile mutant
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Source:
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Phanerochaete chrysosporium. White-rot fungus. Organism_taxid: 5306. Strain: k-3. Expressed in: pichia pastoris. Expression_system_taxid: 4922.
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Resolution:
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2.12Å
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R-factor:
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0.163
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R-free:
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0.220
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Authors:
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J.Vasur,R.Kawai,E.Andersson,G.Widmalm,K.H.M.Jonsson,H.Hansson, A.Engstrom,E.Einarsson,Z.Forsberg,K.Igarashi,M.Sandgren,M.Samejima, J.Stahlberg
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Key ref:
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J.Vasur
et al.
(2010).
Synthesis of cyclic beta-glucan using laminarinase 16A glycosynthase mutant from the basidiomycete Phanerochaete chrysosporium.
J Am Chem Soc,
132,
1724-1730.
PubMed id:
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Date:
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09-Jul-09
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Release date:
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26-Jan-10
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PROCHECK
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Headers
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References
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Q874E3
(Q874E3_PHACH) -
Putative laminarinase from Phanerodontia chrysosporium
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Seq:
Struc:
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318 a.a.
298 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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CATH domain |
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*
PDB and UniProt seqs differ
at 1 residue position (black
cross)
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Enzyme class:
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E.C.3.2.1.6
- endo-1,3(4)-beta-glucanase.
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Reaction:
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Endohydrolysis of 1,3- or 1,4-linkages in beta-D-glucans when the glucose residue whose reducing group is involved in the linkage to be hydrolyzed is itself substituted at C-3.
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J Am Chem Soc
132:1724-1730
(2010)
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PubMed id:
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Synthesis of cyclic beta-glucan using laminarinase 16A glycosynthase mutant from the basidiomycete Phanerochaete chrysosporium.
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J.Vasur,
R.Kawai,
K.H.Jonsson,
G.Widmalm,
A.Engström,
M.Frank,
E.Andersson,
H.Hansson,
Z.Forsberg,
K.Igarashi,
M.Samejima,
M.Sandgren,
J.Ståhlberg.
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ABSTRACT
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Glycosynthases are precise molecular instruments for making specifically linked
oligosaccharides. X-ray crystallography screening of ligands bound to the
1,3(4)-beta-D-glucanase nucleophile mutant E115S of Phanerochaete chrysosporium
Laminarinase 16A (Lam16A) showed that laminariheptaose (L7) bound in an arch
with the reducing and nonreducing ends occupying either side of the catalytic
cleft of the enzyme. The X-ray structure of Lam16A E115S in complex with
alpha-laminariheptaosyl fluoride (alphaL7F) revealed how alphaL7F could make a
nucleophilic attack upon itself. Indeed, when Lam16A E115S was allowed to react
with alphaL7F the major product was a cyclic beta-1,3-heptaglucan, as shown by
mass spectrometry. NMR confirmed uniquely beta-1,3-linkages and no reducing end.
Molecular dynamics simulations indicate that the cyclic laminariheptaose
molecule is not completely planar and that torsion angles at the glycosidic
linkages fluctuate between two energy minima. This is the first report of a
glycosynthase that joins the reducing and nonreducing ends of a single
oligosaccharide and the first reported synthesis of cyclic beta-glucan.
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Links
