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* Residue conservation analysis
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Enzyme class:
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E.C.2.4.1.19
- Cyclomaltodextrin glucanotransferase.
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Reaction:
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Degrades starch to cyclodextrins by formation of a 1,4-alpha-D- glucosidic bond.
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Gene Ontology (GO) functional annotation
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Cellular component
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extracellular region
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1 term
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Biological process
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carbohydrate metabolic process
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1 term
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Biochemical function
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catalytic activity
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9 terms
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DOI no:
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Biochemistry
39:7772-7780
(2000)
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PubMed id:
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Structures of maltohexaose and maltoheptaose bound at the donor sites of cyclodextrin glycosyltransferase give insight into the mechanisms of transglycosylation activity and cyclodextrin size specificity.
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J.C.Uitdehaag,
G.J.van Alebeek,
B.A.van Der Veen,
L.Dijkhuizen,
B.W.Dijkstra.
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ABSTRACT
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The enzymes from the alpha-amylase family all share a similar alpha-retaining
catalytic mechanism but can have different reaction and product specificities.
One family member, cyclodextrin glycosyltransferase (CGTase), has an uncommonly
high transglycosylation activity and is able to form cyclodextrins. We have
determined the 2.0 and 2.5 A X-ray structures of E257A/D229A CGTase in complex
with maltoheptaose and maltohexaose. Both sugars are bound at the donor subsites
of the active site and the acceptor subsites are empty. These structures mimic a
reaction stage in which a covalent enzyme-sugar intermediate awaits binding of
an acceptor molecule. Comparison of these structures with CGTase-substrate and
CGTase-product complexes reveals three different conformational states for the
CGTase active site that are characterized by different orientations of the
centrally located residue Tyr 195. In the maltoheptaose and
maltohexaose-complexed conformation, CGTase hinders binding of an acceptor sugar
at subsite +1, which suggests an induced-fit mechanism that could explain the
transglycosylation activity of CGTase. In addition, the maltoheptaose and
maltohexaose complexes give insight into the cyclodextrin size specificity of
CGTases, since they precede alpha-cyclodextrin (six glucoses) and
beta-cyclodextrin (seven glucoses) formation, respectively. Both ligands show
conformational differences at specific sugar binding subsites, suggesting that
these determine cyclodextrin product size specificity, which is confirmed by
site-directed mutagenesis experiments.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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Z.Li,
M.Wang,
F.Wang,
Z.Gu,
G.Du,
J.Wu,
and
J.Chen
(2007).
gamma-Cyclodextrin: a review on enzymatic production and applications.
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Appl Microbiol Biotechnol, 77,
245-255.
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K.Hirano,
T.Ishihara,
S.Ogasawara,
H.Maeda,
K.Abe,
T.Nakajima,
and
Y.Yamagata
(2006).
Molecular cloning and characterization of a novel gamma-CGTase from alkalophilic Bacillus sp.
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Appl Microbiol Biotechnol, 70,
193-201.
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K.S.Bak-Jensen,
G.André,
T.E.Gottschalk,
G.Paës,
V.Tran,
and
B.Svensson
(2004).
Tyrosine 105 and threonine 212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of barley alpha-amylase 1.
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J Biol Chem, 279,
10093-10102.
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M.Akita,
Y.Hatada,
Y.Hidaka,
Y.Ohta,
M.Takada,
Y.Nakagawa,
K.Ogawa,
T.Nakakuki,
S.Ito,
and
K.Horikoshi
(2004).
Crystallization and preliminary X-ray study of gamma-type cyclodextrin glycosyltransferase from Bacillus clarkii.
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Acta Crystallogr D Biol Crystallogr, 60,
586-587.
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H.Leemhuis,
B.W.Dijkstra,
and
L.Dijkhuizen
(2003).
Thermoanaerobacterium thermosulfurigenes cyclodextrin glycosyltransferase.
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Eur J Biochem, 270,
155-162.
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H.Mori,
K.S.Bak-Jensen,
and
B.Svensson
(2002).
Barley alpha-amylase Met53 situated at the high-affinity subsite -2 belongs to a substrate binding motif in the beta-->alpha loop 2 of the catalytic (beta/alpha)8-barrel and is critical for activity and substrate specificity.
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Eur J Biochem, 269,
5377-5390.
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L.K.Skov,
O.Mirza,
D.Sprogøe,
I.Dar,
M.Remaud-Simeon,
C.Albenne,
P.Monsan,
and
M.Gajhede
(2002).
Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity.
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J Biol Chem, 277,
47741-47747.
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PDB codes:
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M.C.Abad,
K.Binderup,
J.Rios-Steiner,
R.K.Arni,
J.Preiss,
and
J.H.Geiger
(2002).
The X-ray crystallographic structure of Escherichia coli branching enzyme.
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J Biol Chem, 277,
42164-42170.
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PDB code:
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N.Rashid,
J.Cornista,
S.Ezaki,
T.Fukui,
H.Atomi,
and
T.Imanaka
(2002).
Characterization of an archaeal cyclodextrin glucanotransferase with a novel C-terminal domain.
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J Bacteriol, 184,
777-784.
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J.C.Uitdehaag,
B.A.van der Veen,
L.Dijkhuizen,
R.Elber,
and
B.W.Dijkstra
(2001).
Enzymatic circularization of a malto-octaose linear chain studied by stochastic reaction path calculations on cyclodextrin glycosyltransferase.
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Proteins, 43,
327-335.
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Y.Terada,
H.Sanbe,
T.Takaha,
S.Kitahata,
K.Koizumi,
and
S.Okada
(2001).
Comparative study of the cyclization reactions of three bacterial cyclomaltodextrin glucanotransferases.
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Appl Environ Microbiol, 67,
1453-1460.
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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.
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Links
