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Information on EC 2.4.1.245 - alpha,alpha-trehalose synthase and Organism(s) Pyrococcus horikoshii and UniProt Accession O58762
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2.4.1.245 alpha,alpha-trehalose synthaseIUBMB Comments
Requires Mg2+ for maximal activity . The enzyme-catalysed reaction is reversible . In the reverse direction to that shown above, the enzyme is specific for alpha,alpha-trehalose as substrate, as it cannot use alpha- or beta-paranitrophenyl glucosides, maltose, sucrose, lactose or cellobiose . While the enzymes from the thermophilic bacterium Rubrobacter xylanophilus and the hyperthermophilic archaeon Pyrococcus horikoshii can use ADP-, UDP- and GDP-alpha-D-glucose to the same extent [2,3], that from the hyperthermophilic archaeon Thermococcus litoralis has a marked preference for ADP-alpha-D-glucose and that from the hyperthermophilic archaeon Thermoproteus tenax has a marked preference for UDP-alpha-D-glucose .
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The taxonomic range for the selected organisms is: Pyrococcus horikoshii
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
trehalose glycosyltransferring synthase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
NDP-alpha-D-glucose + D-glucose = alpha,alpha-trehalose + NDP
reaction mechanism, overview. The acceptor binding site of TreT shows a wide and commodious groove and lacks the long flexible loop that plays a gating role in ligand binding in trehalose phosphate synthase, TPS, EC 2.4.1.15. A wide space at the fissure between two domains and the relative shift of the N-domain in one of the crystal forms suggest that an interactive conformational change between two domains would occur, allowing a more compact architecture for catalysis
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
NDP-alpha-D-glucose:D-glucose 1-alpha-D-glucosyltransferase
Requires Mg2+ for maximal activity [1]. The enzyme-catalysed reaction is reversible [1]. In the reverse direction to that shown above, the enzyme is specific for alpha,alpha-trehalose as substrate, as it cannot use alpha- or beta-paranitrophenyl glucosides, maltose, sucrose, lactose or cellobiose [1]. While the enzymes from the thermophilic bacterium Rubrobacter xylanophilus and the hyperthermophilic archaeon Pyrococcus horikoshii can use ADP-, UDP- and GDP-alpha-D-glucose to the same extent [2,3], that from the hyperthermophilic archaeon Thermococcus litoralis has a marked preference for ADP-alpha-D-glucose [1] and that from the hyperthermophilic archaeon Thermoproteus tenax has a marked preference for UDP-alpha-D-glucose [4].
CAS REGISTRY NUMBER
COMMENTARY
126341-88-6
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
-
-
-
?
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-trehalose
-
-
-
r
UDP-alpha-D-glucose + D-galactose
alpha-D-Glc-(1->1)-alpha-D-Glc + UDP
-
-
-
?
UDP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + UDP
-
-
-
?
ADP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + ADP
-
GDP-alpha-D-glucose is the most favored in terms of reaction specificity, kcat/Km. UDP-alpha-D-glucose and ADP-alpha-D-glucose are employed with less preferences. The enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
alpha,alpha-trehalose + ADP
ADP-alpha-D-glucose + D-glucose
-
the enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
alpha,alpha-trehalose + GDP
GDP-alpha-D-glucose + D-glucose
-
the enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
alpha,alpha-trehalose + UDP
UDP-alpha-D-glucose + D-glucose
-
the enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
GDP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + GDP
-
GDP-alpha-D-glucose is the most favored in terms of reaction specificity, kcat/Km. UDP-alpha-D-glucose and ADP-alpha-D-glucose are employed with less preferences. The enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
UDP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + UDP
-
GDP-alpha-D-glucose is the most favored in terms of reaction specificity, kcat/Km. UDP-alpha-D-glucose and ADP-alpha-D-glucose are employed with less preferences. The enzyme reversely cleaves alpha,alpha-trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-alpha-D-glucose. Although ADP-alpha-D-glucose is the least favorable donor, the counterpart, ADP, is the most favorable acceptor for the reverse synthesis of NDP-alpha-D-glucose in kcat/Km. GDP and UDP are less preferred, compared to ADP
-
-
r
additional information
?
-
UDP-glucose is as efficient as ADP-glucose and GDP-glucose
-
-
?
additional information
?
-
molecular basis of the synthetic mechanism of trehalose, or the nucleotide sugar in the reverse reaction of TreT. TreT can utilize ADP-glucose and GDP-glucose as well as UDP-glucose to synthesize trehalose, the nucleotide sugar molecule is recognized by the nucleotide-sensing Gly-Gly-Leu motif that is located at the domain interface in many GT-B family members, and TreT contains this motif at residues 52-55, the acceptor molecule binds predominantly to the N-terminal domain
-
-
?
additional information
?
-
-
molecular basis of the synthetic mechanism of trehalose, or the nucleotide sugar in the reverse reaction of TreT. TreT can utilize ADP-glucose and GDP-glucose as well as UDP-glucose to synthesize trehalose, the nucleotide sugar molecule is recognized by the nucleotide-sensing Gly-Gly-Leu motif that is located at the domain interface in many GT-B family members, and TreT contains this motif at residues 52-55, the acceptor molecule binds predominantly to the N-terminal domain
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-trehalose
-
-
-
r
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
the acceptor binding site of TreT shows a wide and commodious groove and lacks the long flexible loop that plays a gating role in ligand binding in trehalose phosphate synthase, TPS, active site, and donor and acceptor binding pocket structure, overview. A wide space at the fissure between two domains and the relative shift of the N-domain in one of the crystal forms suggest that an interactive conformational change between two domains would occur, allowing a more compact architecture for catalysis
additional information
-
the acceptor binding site of TreT shows a wide and commodious groove and lacks the long flexible loop that plays a gating role in ligand binding in trehalose phosphate synthase, TPS, active site, and donor and acceptor binding pocket structure, overview. A wide space at the fissure between two domains and the relative shift of the N-domain in one of the crystal forms suggest that an interactive conformational change between two domains would occur, allowing a more compact architecture for catalysis
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
48197
x * 48197, calculated, x * 49871, MALDI-TOF mass spectrometry of recombinant His-tagged protein
49871
x * 48197, calculated, x * 49871, MALDI-TOF mass spectrometry of recombinant His-tagged protein
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 48197, calculated, x * 49871, MALDI-TOF mass spectrometry of recombinant His-tagged protein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified enzyme free or as TreT-UDP binary complex, 10 mg/ml native enzyme from PEG 3350 25%, 0.2 M MgCl2, and 0.1 M sodium HEPES, 18°C, the selenomethionine-substituted protein crystal grow from 21% methoxy PEG 2000, 0.18 M ammonium sulfate, and 0.1 M sodium acetate, pH 4.6, for the UDP-glucose complex crystal, 5 mM UDPG is added to 10 mg/ml E326A protein for 1 h prior to setup of the crystallization using the same conditions as for the native crystal, X-ray diffraction structure determination and analysis at 2.3-3.0 A resolution, single-wavelength anomalous dispersion phasing
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D134E
the mutant shows 10% reduced activity in the forward reaction, and 50% reduced activity in the reverse reaction
F85R
the mutant shows 95% reduced activity in the forward reaction, and no activity in the reverse reaction
F85Y
the mutant shows 10% reduced activity in the forward reaction, and 40% reduced activity in the reverse reaction
H155D
inactive in the forward reaction, 80% reduced activity in the reverse reaction compared to wild-type
H92A
the mutant shows wild-type activity in the forward reaction, and 90% reduced activity in the reverse reaction
Q96A
the mutant shows 10% reduced activity in the forward reaction, and 90% reduced activity in the reverse reaction
R239A
inactive in the reverse reaction, 10% reduced activity in the forward reaction compared to wild-type
T49H
the mutant shows 90% reduced activity in the forward reaction, and 98% reduced activity in the reverse reaction
V309L
the mutant shows wild-type activity in the forward reaction, and 25% reduced activity in the reverse reaction
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene treT, DNA and amino acid sequence determination and analysis, sequence comparison
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
synthesis
-
the enzyme may be useful for the regeneration of NDP-alpha-D-glucose from NDP, especially for ADP-alpha-D-glucose from ADP, with trehalose as a glucose resource
synthesis
trehalose synthase from Pyrococcus horikoshii can be applied to the sugar nucleotide cycling process for the synthesis of functional alpha-galactosyl oligosaccharides, alpha-galactose epitopes and globotriose, using the effective regeneration of UDP-galactose
synthesis
synthesis of the trehalase analogue alpha-D-Glc-(1->1)-alpha-D-Glc, that might be effective at inhibiting intestinal brush-order disaccharidases
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ryu, S.I.; Park, C.S.; Cha, J.; Woo, E.J.; Lee, S.B.
A novel trehalose-synthesizing glycosyltransferase from Pyrococcus horikoshii: Molecular cloning and characterization
Biochem. Biophys. Res. Commun.
329
429-436
2005
Pyrococcus horikoshii (O58762)
Ryu, S.I.; Woo, J.B.; Lee, S.B.
Coupling reactions of trehalose synthase from Pyrococcus horikoshii: Cost-effective synthesis and anti-adhesive activity of beta-galactosyl oligosaccharides using a one-pot three-enzyme system with trehalose
Biores. Technol.
136
743-746
2013
Pyrococcus horikoshii (O58762)
Woo, E.J.; Ryu, S.I.; Song, H.N.; Jung, T.Y.; Yeon, S.M.; Lee, H.A.; Park, B.C.; Park, K.H.; Lee, S.B.
Structural insights on the new mechanism of trehalose synthesis by trehalose synthase TreT from Pyrococcus horikoshii
J. Mol. Biol.
404
247-259
2010
Pyrococcus horikoshii (O58762), Pyrococcus horikoshii
Ryu, S.; Kim, J.; Kim, E.; Chung, S.; Lee, S.
Catalytic reversibility of Pyrococcus horikoshii trehalose synthase: Efficient synthesis of several nucleoside diphosphate glucoses with enzyme recycling
Process Biochem.
46
128-134
2011
Pyrococcus horikoshii
-
Kim, H.; Chang, Y.; Ryu, S.; Moon, S.; Lee, S.
Enzymatic synthesis of a galactose-containing trehalose analogue disaccharide by Pyrococcus horikoshii trehalose-synthesizing glycosyltransferase Inhibitory effects on several disaccharidase activities
J. Mol. Catal. B
49
98-103
2007
Pyrococcus horikoshii (O58762), Pyrococcus horikoshii OT-3 (O58762)
-
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