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Information on EC 2.6.1.33 - dTDP-4-amino-4,6-dideoxy-D-glucose transaminase and Organism(s) Escherichia coli and UniProt Accession P27833

for references in articles please use BRENDA:EC2.6.1.33
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IUBMB Comments
A pyridoxal-phosphate protein.
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This record set is specific for:
Escherichia coli
UNIPROT: P27833
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
sugar aminotransferase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminotransferase, thymidine diphospho-4-amino-4,6-dideoxyglucose
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sugar aminotransferase
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TDP-4-keto-6-deoxy-D-glucose transaminase
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thymidine diphospho-4-amino-4,6-dideoxyglucose aminotransferase
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thymidine diphospho-4-amino-6-deoxyglucose aminotransferase
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thymidine diphospho-4-keto-6-deoxy-D-glucose transaminase
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thymidine diphospho-4-keto-6-deoxy-D-glucose-glutamic transaminase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dTDP-4-amino-4,6-dideoxy-alpha-D-glucose + 2-oxoglutarate = dTDP-4-dehydro-6-deoxy-alpha-D-glucose + L-glutamate
show the reaction diagram
substrate specificity might be related to conserved motifs VLalpha, VLbeta, and VLgamma, around the active site
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amino group transfer
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SYSTEMATIC NAME
IUBMB Comments
dTDP-4-amino-4,6-dideoxy-D-glucose:2-oxoglutarate aminotransferase
A pyridoxal-phosphate protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-19-2
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dTDP-4-dehydro-6-deoxy-D-glucose + L-alanine
dTDP-4-amino-4,6-dideoxy-D-galactose + pyruvate
show the reaction diagram
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L-alanine, poor amino donor
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?
dTDP-4-dehydro-6-deoxy-D-glucose + L-glutamate
dTDP-4-amino-4,6-dideoxy-D-galactose + 2-oxoglutarate
show the reaction diagram
dTDP-4-dehydro-6-deoxy-D-glucose + L-glutamate
dTDP-4-amino-4,6-dideoxy-D-glucose + 2-oxoglutarate
show the reaction diagram
dTDP-4-dehydro-6-deoxy-D-glucose + L-glutamine
dTDP-4-amino-4,6-dideoxy-D-galactose + 2-oxoglutamine
show the reaction diagram
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co-substrate L-glutamine, 15.9% conversion rate
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?
dTDP-4-dehydro-6-deoxy-D-mannose + L-glutamate
dTDP-4-amino-6-deoxy-D-mannose + 2-oxoglutarate
show the reaction diagram
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?
TDP-4-amino-4,6-dideoxy-alpha-D-glucose + 2-oxoglutarate
TDP-4-dehydro-6-deoxy-alpha-D-glucose + L-glutamate
show the reaction diagram
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?
additional information
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
dTDP-4-dehydro-6-deoxy-D-glucose + L-glutamate
dTDP-4-amino-4,6-dideoxy-D-glucose + 2-oxoglutarate
show the reaction diagram
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r
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(aminooxy)acetic acid
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complete inhibition by a mixture of hydroxylamine, (aminooxy)acetic acid, and gabaculine as inhibitors specific for pyridoxal 5'-phosphate-dependent enzymes
Co2+
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20% conversion in presence of Co2+
gabaculine
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complete inhibition by a mixture of hydroxylamine, (aminooxy)acetic acid, and gabaculine as inhibitors specific for pyridoxal 5'-phosphate-dependent enzymes
hydroxylamine
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complete inhibition by a mixture of hydroxylamine, (aminooxy)acetic acid, and gabaculine as inhibitors specific for pyridoxal 5'-phosphate-dependent enzymes
additional information
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not inhibitory: Mg2+, EDTA
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.22
2-oxoglutarate
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38°C, pH 7.0-7.6
0.0458
dTDP-4-dehydro-6-deoxy-D-glucose
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pH 7.4, 37°C
5.1
L-glutamate
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38°C, pH 7.0-7.6
0.032
TDP-4-amino-4,6-dideoxy-alpha-D-glucose
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38°C, pH 7.0-7.6
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0.14
TDP-4-keto-6-deoxy-D-glucose
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38°C, pH 7.0-7.6
additional information
additional information
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kinetics in a coupled assay
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.66
dTDP-4-dehydro-6-deoxy-D-glucose
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pH 7.4, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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activity measurement in a coupled assay with NADH-dependent L-GDH
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9.5
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enzyme shows over 80% of maximal activity at pH 6.0 and pH 8.0, and very low activity below pH 5.0 and above pH 9.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 45
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25% of maximal activity at 45°C
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
180000
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recombinant His6-tagged enzyme, gel filtration
44000
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4 * 44000, recombinant His6-tagged enzyme, SDS-PAGE, 4 * 44452, recombinant His6-tagged enzyme, nucleotide sequence calculation, 4 * 44465, recombinant His6-tagged enzyme, MALDI-TOF mass spectrometry
44452
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4 * 44000, recombinant His6-tagged enzyme, SDS-PAGE, 4 * 44452, recombinant His6-tagged enzyme, nucleotide sequence calculation, 4 * 44465, recombinant His6-tagged enzyme, MALDI-TOF mass spectrometry
44465
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4 * 44000, recombinant His6-tagged enzyme, SDS-PAGE, 4 * 44452, recombinant His6-tagged enzyme, nucleotide sequence calculation, 4 * 44465, recombinant His6-tagged enzyme, MALDI-TOF mass spectrometry
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
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4 * 44000, recombinant His6-tagged enzyme, SDS-PAGE, 4 * 44452, recombinant His6-tagged enzyme, nucleotide sequence calculation, 4 * 44465, recombinant His6-tagged enzyme, MALDI-TOF mass spectrometry
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0°C or -15°C, partially purified enzyme, stable for several weeks
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0°C, crude cell extract, stable for several days
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
partial from strain B, 28fold
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recombinant C-terminally His6-tagged enzyme from strain BL21(DE3) by nickel chelate and hydrophobic interaction chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression with His-tag
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gene WecC, amino acid sequence determination and analysis, overexpression of C-terminally His6-tagged enzyme in strain BL21(DE3), phylogenetic analysis
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
engineering of Escherichia coli to divert the flow of carbon flux from glucose-1-phosphate to thymidine diphosphate 4-keto 4,6-dideoxy-D-glucose (dTKDG), an intermediate of various dTDP-sugars. Glucose phosphate isomerase, glucose-6-phosphate dehydrogenase and uridylyltransferase genes are deleted while dTDP-D-glucose synthase and dTDP-Dglucose 4,6-dehydratase are overexpressed to produce a pool of dTKDG in the cell cytosol. The flow of dTKDG is further diverted to dTDP-D-viosamine, dTDP 4-amino 4,6-dideoxy-D-galactose, and dTDP 3-amino 3,6-dideoxy-D-galactose sugars using sugar aminotransferases gerB, wecE, and fdtB, respectively, from different sources. These sugar moieties are transferred to the 3-hydroxyl position of quercetin and kaempferol with the help of Arabidopsis thaliana glycosyltransferase ArGT3
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Matsuhashi, M.; Strominger, J.L.
Thymidine diphosphate 4-acetamido-2,6-dideoxyhexoses. 3. Purification and properties of thymidine diphosphate 4-keto-6-deoxy-D-glucose transaminase from Escherichia coli strain B
J. Biol. Chem.
241
4738-4744
1966
Escherichia coli, Escherichia coli B / ATCC 11303
Manually annotated by BRENDA team
Hwang, B.Y.; Lee, H.J.; Yang, Y.H.; Joo, H.S.; Kim, B.G.
Characterization and investigation of substrate specificity of the sugar aminotransferase WecE from E. coli K12
Chem. Biol.
11
915-925
2004
Escherichia coli
Manually annotated by BRENDA team
Wang, Y.; Xu, Y.; Perepelov, A.V.; Qi, Y.; Knirel, Y.A.; Wang, L.; Feng, L.
Biochemical characterization of dTDP-D-Qui4N and dTDP-D-Qui4NAc biosynthetic pathways in Shigella dysenteriae type 7 and Escherichia coli O7
J. Bacteriol.
189
8626-8635
2007
Escherichia coli, Escherichia coli O7, Shigella dysenteriae
Manually annotated by BRENDA team
Pandey, R.; Parajuli, P.; Chu, L.; Darsandhari, S.; Sohng, J.
Biosynthesis of amino deoxy-sugar-conjugated flavonol glycosides by engineered Escherichia coli
Biochem. Eng. J.
101
191-199
2015
Escherichia coli (P27833)
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Manually annotated by BRENDA team