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Information on EC 2.4.1.245 - alpha,alpha-trehalose synthase
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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 expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
trehalose glycosyltransferring synthase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
PH1035
TmTreT
trehalose glycosyltransferring synthase
TreT
trehalose glycosyltransferring synthase
Thermoproteus tenax Kra 1 / DSM 2078
-
-
-
TreT
Thermoproteus tenax Kra 1 / DSM 2078
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
NDP-alpha-D-glucose + D-glucose = alpha,alpha-trehalose + NDP
-
-
-
-
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
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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PATHWAY SOURCE
PATHWAYS
BRENDA
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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].
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CAS REGISTRY NUMBER
COMMENTARY
126341-88-6
-
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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-trehalose
-
-
-
r
alpha,alpha-trehalose + CDP
CDP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
r
UDP-alpha-D-glucose + D-fructose
?
-
the enzyme employs a glucose acceptor with the highest preference and other monosaccharides (D-fructose, D-mannose) with less affinities acceptors. The trehalose analogue containing mannose is produced with approximately 70% of trehalose production and a two-fold greater amount than the analogue containing a fructose for the 12-24 h reaction
-
-
r
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
-
-
-
?
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
100% activity
-
-
?
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
-
-
-
-
r
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
-
10% activity with ADP-glucose compared to UDP-glucose
-
-
r
ADP-alpha-D-glucose + D-glucose
ADP + alpha,alpha-1,1-trehalose
Thermoproteus tenax Kra 1 / DSM 2078
-
10% activity with ADP-glucose compared to UDP-glucose
-
-
r
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
ADP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + 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 + ADP
ADP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
r
alpha,alpha-trehalose + ADP
ADP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
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 + GDP
GDP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
?
alpha,alpha-trehalose + GDP
GDP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
?
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
alpha,alpha-trehalose + UDP
UDP-alpha-D-glucose + D-glucose
-
both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
?
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
GDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + GDP
48.2% activity compared to ADP-glucose
-
-
?
GDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + GDP
-
5% of the efficiency with ADP-glucose
-
-
r
UDP-alpha-D-glucose + D-galactose
alpha-D-Glc-(1->1)-alpha-D-Glc + UDP
-
-
-
?
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
-
-
-
?
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
UDP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + UDP
-
-
-
?
UDP-alpha-D-glucose + D-glucose
alpha,alpha-trehalose + UDP
-
the purified soluble enzyme, as well as the glutathione-S-transferase-fusion enzyme, show the catalytic specificity for synthesizing trehalose from UDP-alpha-D-glucose as the donor and glucose as the acceptor. The enzyme employs a glucose acceptor with the highest preference and other monosaccharides (D-fructose, D-mannose) with less affinities acceptors. Both soluble and glutathione-S-transferase-fusion enzymes have the greatest preference for ADP as the acceptor substrate in the synthesis of NDP-Glc, and they also employ GDP, UDP, and CDP as less favorable substrates, approximately in this order
-
-
r
UDP-alpha-D-glucose + D-mannose
?
-
the enzyme employs a glucose acceptor with the highest preference and other monosaccharides (D-fructose, D-mannose) with less affinities acceptors. The trehalose analogue containing mannose is produced with approximately 70% of trehalose production and a two-fold greater amount than the analogue containing a fructose for the 12-24 h reaction
-
-
r
UDP-alpha-D-glucose + D-mannose
?
-
the enzyme employs a glucose acceptor with the highest preference and other monosaccharides (D-fructose, D-mannose) with less affinities acceptors. The trehalose analogue containing mannose is produced with approximately 70% of trehalose production and a two-fold greater amount than the analogue containing a fructose for the 12-24 h reaction
-
-
r
UDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + UDP
32.7% activity compared to ADP-glucose
-
-
?
UDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + UDP
-
6% of the efficiency with ADP-glucose
-
-
r
UDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + UDP
-
clear preference for UDP-glucose over ADP-glucose
-
-
r
UDP-glucose + D-glucose
alpha,alpha-1,1-trehalose + UDP
Thermoproteus tenax Kra 1 / DSM 2078
-
clear preference for UDP-glucose over ADP-glucose
-
-
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
-
-
?
additional information
?
-
-
reaction is reversible, formation of trehalose is favored by rate of reaction and equilibrium. No substrate: glucose 1-phosphate, glucose 6-phosphate, fructose, mannose, galactose, xylose, 2-deoxyglucose, glucosamine, alpha-methylglucoside, sorbitol
-
-
?
additional information
?
-
-
no activity with maltose and glucose 1-phosphate
-
-
?
additional information
?
-
Thermoproteus tenax Kra 1 / DSM 2078
-
no activity with maltose and glucose 1-phosphate
-
-
?
additional information
?
-
-
the enzyme does not utilize galactose as an acceptor
-
-
?
additional information
?
-
-
the enzyme does not utilize galactose as an acceptor
-
-
?
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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
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Mg2+
Mn2+
additional information
no enhanced activity in the presence of 20 mM Fe3+ or Mn2+
Mg2+
Mg2+ is required for activity, 20 mM Mg+ induces slightly enhanced activity
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.23
UDP-glucose
-
in 50 mM HEPES/KOH (pH 7.0), with 20 mM MgCl2, at 70°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5 - 8
-
pH 4.5: about 45% of maximal activity, pH 8.0: about 40% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
60
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
35 - 70
-
35°C: about 40% of maximal activity, 70°C: about 70% of maximal activity
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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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
Show AA Sequence and Transmembrane Helices (320 entries)
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Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
47000
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
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 48197, calculated, x * 49871, MALDI-TOF mass spectrometry of recombinant His-tagged protein
monomer
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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
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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
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
-
2 h, activity of the the glutathione-S-transferase-fusion enzyme is fully maintained
60
-
2 h, 10% decrease of the activity of the the glutathione-S-transferase-fusion enzyme, activity of the soluble enzyme is decreased by 35%
60 - 70
the half-lives for inactivation at 60°C and at 70°C are 309 h and 4.1 h, respectively
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE