BRENDA - Enzyme Database
show all sequences of 4.2.1.46

A 96-well microtiter plate assay for high-throughput screening of Mycobacterium tuberculosis dTDP-D-glucose 4,6-dehydratase inhibitors

Shi, X.; Sha, S.; Liu, L.; Li, X.; Ma, Y.; Anal. Biochem. 498, 53-58 (2016)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
NAD+
during RmlB enzyme turnover, NAD+ extracts a hydride from the C40 atom of dTDP-D-glucose and forms NADH. After the elimination of water, the hydride is then returned to the C60 atom of dTDP-4-dehydro-5,6-glucosene intermediate and NADH regenerates NAD+. The optimal concentration of NAD+ is between 0.01 mM and 0.02 mM
Mycobacterium tuberculosis
Application
Application
Commentary
Organism
analysis
an absorbance-based microtiter plate assay is developed for RmlB activity. It can be used for high-throughput screening of RmlB inhibitors
Mycobacterium tuberculosis
pharmacology
the enzyme is essential to mycobacterial growth and is not found in humans, therefore, it is a potential target for developing new anti-tuberculosis drugs
Mycobacterium tuberculosis
Cloned(Commentary)
Cloned (Commentary)
Organism
expression in Escherichia coli BL21(DE3)
Mycobacterium tuberculosis
Inhibitors
Inhibitors
Commentary
Organism
Structure
Ca2+
0.5 mM, about 50% loss of activity
Mycobacterium tuberculosis
dTDP
competitive inhibitor
Mycobacterium tuberculosis
dTTP
competitive inhibitor
Mycobacterium tuberculosis
K+
1 mM, about 10% loss of activity
Mycobacterium tuberculosis
Mg2+
0.5 mM, about 30% loss of activity
Mycobacterium tuberculosis
Mn2+
0.2 mM, about 50% loss of activity
Mycobacterium tuberculosis
Na+
1 mM, about 25% loss of activity
Mycobacterium tuberculosis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.05
-
dTDP-alpha-D-glucose
pH 7.5, 30°C
Mycobacterium tuberculosis
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
metal ion is not required for RmlB enzymatic reaction. Stimulated by Mg2+ at the low concentration (0.1 mM) but shows different levels of inhibition by increased Mg2+ concentration
Mycobacterium tuberculosis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
39000
-
SDS-PAGE
Mycobacterium tuberculosis
Organism
Organism
UniProt
Commentary
Textmining
Mycobacterium tuberculosis
P9WN65
-
-
Mycobacterium tuberculosis H37Rv
P9WN65
-
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
dTDP-alpha-D-glucose
-
746731
Mycobacterium tuberculosis
dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
-
-
?
dTDP-alpha-D-glucose
-
746731
Mycobacterium tuberculosis H37Rv
dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
RmlB
-
Mycobacterium tuberculosis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
-
Mycobacterium tuberculosis
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
26
60
25°C: about 55% of maximal activity, 60°C: about 40% of maximal activity
Mycobacterium tuberculosis
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
466.6
-
dTDP-alpha-D-glucose
pH 7.5, 30°C
Mycobacterium tuberculosis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
Hepes buffer
Mycobacterium tuberculosis
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.5
8.5
pH 6.5: about 45% of maximal activity, pH 8.5: about 45% of maximal activity, HEPES buffer
Mycobacterium tuberculosis
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00267
-
dTDP
pH 7.5, 30°C
Mycobacterium tuberculosis
0.00567
-
dTTP
pH 7.5, 30°C
Mycobacterium tuberculosis
IC50 Value
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.006
-
pH 7.5, 30°C
Mycobacterium tuberculosis
dTDP
0.032
-
pH 7.5, 30°C
Mycobacterium tuberculosis
dTTP
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
NAD+
during RmlB enzyme turnover, NAD+ extracts a hydride from the C40 atom of dTDP-D-glucose and forms NADH. After the elimination of water, the hydride is then returned to the C60 atom of dTDP-4-dehydro-5,6-glucosene intermediate and NADH regenerates NAD+. The optimal concentration of NAD+ is between 0.01 mM and 0.02 mM
Mycobacterium tuberculosis
Application (protein specific)
Application
Commentary
Organism
analysis
an absorbance-based microtiter plate assay is developed for RmlB activity. It can be used for high-throughput screening of RmlB inhibitors
Mycobacterium tuberculosis
pharmacology
the enzyme is essential to mycobacterial growth and is not found in humans, therefore, it is a potential target for developing new anti-tuberculosis drugs
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli BL21(DE3)
Mycobacterium tuberculosis
IC50 Value (protein specific)
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.006
-
pH 7.5, 30°C
Mycobacterium tuberculosis
dTDP
0.032
-
pH 7.5, 30°C
Mycobacterium tuberculosis
dTTP
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
Ca2+
0.5 mM, about 50% loss of activity
Mycobacterium tuberculosis
dTDP
competitive inhibitor
Mycobacterium tuberculosis
dTTP
competitive inhibitor
Mycobacterium tuberculosis
K+
1 mM, about 10% loss of activity
Mycobacterium tuberculosis
Mg2+
0.5 mM, about 30% loss of activity
Mycobacterium tuberculosis
Mn2+
0.2 mM, about 50% loss of activity
Mycobacterium tuberculosis
Na+
1 mM, about 25% loss of activity
Mycobacterium tuberculosis
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00267
-
dTDP
pH 7.5, 30°C
Mycobacterium tuberculosis
0.00567
-
dTTP
pH 7.5, 30°C
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.05
-
dTDP-alpha-D-glucose
pH 7.5, 30°C
Mycobacterium tuberculosis
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
metal ion is not required for RmlB enzymatic reaction. Stimulated by Mg2+ at the low concentration (0.1 mM) but shows different levels of inhibition by increased Mg2+ concentration
Mycobacterium tuberculosis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
39000
-
SDS-PAGE
Mycobacterium tuberculosis
Purification (Commentary) (protein specific)
Commentary
Organism
-
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
dTDP-alpha-D-glucose
-
746731
Mycobacterium tuberculosis
dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
-
-
?
dTDP-alpha-D-glucose
-
746731
Mycobacterium tuberculosis H37Rv
dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
-
Mycobacterium tuberculosis
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
26
60
25°C: about 55% of maximal activity, 60°C: about 40% of maximal activity
Mycobacterium tuberculosis
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
466.6
-
dTDP-alpha-D-glucose
pH 7.5, 30°C
Mycobacterium tuberculosis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
Hepes buffer
Mycobacterium tuberculosis
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.5
8.5
pH 6.5: about 45% of maximal activity, pH 8.5: about 45% of maximal activity, HEPES buffer
Mycobacterium tuberculosis
General Information
General Information
Commentary
Organism
metabolism
the enzyme is involved in biosynthesis of dTDP-L-rhamnose, which is a sugar donor to the synthesis of the cell wall linker, D-N-acetylglucosamine-L-rhamnose
Mycobacterium tuberculosis
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme is involved in biosynthesis of dTDP-L-rhamnose, which is a sugar donor to the synthesis of the cell wall linker, D-N-acetylglucosamine-L-rhamnose
Mycobacterium tuberculosis
Other publictions for EC 4.2.1.46
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
747917
Li
Characterization of the dTDP- ...
Campylobacter jejuni
Glycobiology
27
358-369
2017
-
-
1
-
-
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-
-
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1
-
1
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1
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2
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1
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1
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1
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1
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1
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2
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1
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1
-
-
-
-
1
1
-
-
-
746731
Shi
A 96-well microtiter plate as ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Anal. Biochem.
498
53-58
2016
1
2
1
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-
7
1
1
1
1
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4
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1
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2
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1
1
1
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1
1
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1
2
1
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2
7
2
1
1
1
1
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1
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2
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1
1
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1
1
1
-
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1
1
-
-
-
747027
Feng
Identification of a dTDP-rham ...
Caenorhabditis elegans
Biochem. J.
473
1507-1521
2016
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-
1
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1
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6
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1
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1
1
-
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-
749282
Ma
-
Insights into the catalytic m ...
Streptomyces venezuelae
RSC Adv.
4
35449-35458
2014
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1
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1
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730235
Wu
Identification of elaiophylin ...
Streptomyces sp., Streptomyces sp. 7-145
J. Nat. Prod.
76
2153-2157
2013
-
-
-
-
-
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2
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4
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2
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729054
Chen
Distribution of dTDP-glucose-4 ...
Bacillus sp. (in: Bacteria), Bacillus sp. (in: Bacteria) 5187, Dietzia sp. 3149, Dietzia sp. DQ12-45-1b 3149, Ensifer sp. AS08, Ensifer sp. AS08 448, Filomicrobium sp. 454, Glycomyces sp. 3338, Micromonospora sp. 114, Micromonospora sp., Micromonospora sp. 282, Micromonospora sp. 3113, Micromonospora sp. 3134, Micromonospora sp. 3137, Micromonospora sp. 3191, Micromonospora sp. 3372-2, Micromonospora sp. 3387, Micromonospora sp. 3437-1, Micromonospora sp. 5297, Nocardia sp., Nocardia sp. 5314, Prauseria sp. 3425, Pseudomonas sp., Pseudomonas sp. 5302, Rhodococcus sp., Rhodococcus sp. 3376-1, Rhodospirillaceae bacterium 5305, Saccharothrix sp. 5133, Shimazuella sp. 3435, Streptomyces sp., Streptomyces sp. 3127, Streptomyces sp. 3419, Streptomyces sp. 5175, Streptomyces sp. 5191, Streptomyces sp. 5311, Streptomyces sp. 5320, Streptomyces sp. 568, Streptomyces sp. SCC 2136, Streptosporangium sp., Streptosporangium sp. 5322, Verrucosispora sp. 3133
Appl. Microbiol. Biotechnol.
90
1347-1359
2011
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41
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42
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41
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32
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41
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41
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701859
Singh
Precursor for biosynthesis of ...
Streptomyces peucetius
Appl. Microbiol. Biotechnol.
85
1565-1574
2010
-
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1
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1
2
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3
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1
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2
2
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716065
Parakkottil Chothi
Identification of an L-rhamnos ...
Acanthamoeba polyphaga Mimivirus, Acanthocystis turfacea chlorella virus 1
J. Virol.
84
8829-8838
2010
-
-
2
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2
4
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2
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2
2
4
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2
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2
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4
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2
2
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2
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2
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690812
Pfoestl
Biosynthesis of dTDP-3-acetami ...
Thermoanaerobacterium thermosaccharolyticum, Thermoanaerobacterium thermosaccharolyticum E207-71
Biochem. J.
410
187-194
2008
-
-
1
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2
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1
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1
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2
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652536
Allard
High resolution X-ray structur ...
Streptomyces venezuelae
J. Biol. Chem.
279
2211-2220
2004
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1
1
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2
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1
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664694
Du
Identification and functional ...
Streptoalloteichus tenebrarius
Curr. Microbiol.
49
99-107
2004
-
1
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1
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6
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1
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650136
Hegeman
Concerted and stepwise dehydra ...
Escherichia coli
Biochemistry
41
2797-2804
2002
-
-
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2
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2
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1
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1
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652770
Sohng
-
Function of lysine-148 in dTDP ...
Streptomyces antibioticus, Streptomyces antibioticus Tue99
J. Microbiol. Biotechnol.
12
217-221
2002
-
-
-
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1
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1
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1
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650028
Gross
Dehydration is catalyzed by gl ...
Escherichia coli
Biochemistry
40
12497-12504
2001
-
-
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9
-
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2
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2
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19
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1
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1
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9
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2
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19
-
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-
-
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-
650072
Hegeman
Probing catalysis by Escherich ...
Escherichia coli
Biochemistry
40
6598-6610
2001
-
-
-
-
17
-
2
19
-
-
-
-
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2
-
-
1
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2
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20
-
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20
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650089
Gerratana
Mechanistic roles of Thr134, T ...
Escherichia coli
Biochemistry
40
9187-9195
2001
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7
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8
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8
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652851
Allard
The crystal structure of dTDP- ...
Salmonella enterica
J. Mol. Biol.
307
283-295
2001
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1
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649133
Allard
The purification, crystallizat ...
Salmonella enterica
Acta Crystallogr. Sect. D
56
222-225
2000
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1
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649934
Gross
Characterization of enzymatic ...
Escherichia coli
Biochemistry
39
13633-13640
2000
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1
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652771
Yoo
-
Expression of orf7(oxi III) as ...
Streptomyces antibioticus, Streptomyces antibioticus Tu99
J. Microbiol. Biotechnol.
9
206-212
1999
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2
1
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5725
Stein
Combined preparative enzymatic ...
Salmonella enterica subsp. enterica serovar Typhimurium
Glycoconj. J.
15
139-145
1998
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1
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1
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1
-
1
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1
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5724
Naundorf
Substrate specificity of nativ ...
Escherichia coli
Carbohydr. Res.
285
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1996
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1
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3
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3
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3
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3
-
3
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5723
Linton
Cloning of the genes encoding ...
Saccharopolyspora erythraea
Gene
153
33-40
1995
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1
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5726
Thompson
Purification and characterizat ...
Streptomyces peucetius, Streptomyces sp. C5
J. Gen. Microbiol.
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779-786
1992
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6
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4
2
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2
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5722
Romana
High level expression and puri ...
Salmonella enterica subsp. enterica serovar Typhimurium
Biochem. Biophys. Res. Commun.
174
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1991
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3
1
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1
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1
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1
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1
1
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1
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1
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2
1
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1
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1
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3
1
1
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1
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5712
Vara
Purification of thymidine-diph ...
Saccharopolyspora erythraea
J. Biol. Chem.
263
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1988
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1
1
2
1
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1
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1
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1
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2
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2
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2
1
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1
1
2
1
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5713
Matern
Stoffwechselprodukte von Mikro ...
Streptomyces rimosus
Arch. Mikrobiol.
88
37-48
1973
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3
1
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1
2
2
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1
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1
1
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1
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1
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2
2
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1
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1
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5714
Glaser
-
Dehydration in nucleotide-link ...
Escherichia coli
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
5
465-480
1971
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1
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1
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2
1
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1
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1
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1
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4
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3
1
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2
1
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5715
Zarkowsky
The mechanism of 6-deoxyhexose ...
Escherichia coli
J. Biol. Chem.
245
6599-6606
1970
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2
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2
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1
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2
1
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1
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1
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2
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2
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5716
Zarkowsky
The mechanism of 6-deoxyhexose ...
Escherichia coli
Biochem. Biophys. Res. Commun.
38
787-793
1970
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2
1
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2
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1
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1
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2
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1
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1
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5717
Wang
Biological mechanisms involved ...
Escherichia coli
J. Biol. Chem.
245
8-14
1970
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1
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5718
Zarkowsky
The mechanism of 6-deoxyhexose ...
Escherichia coli
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244
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1969
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2
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5719
Wang
Biological mechanisms involved ...
Escherichia coli
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244
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5720
Melo
The mechanism of 6-deoxyhexose ...
Escherichia coli
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243
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1968
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5721
Gilbert
Thymidine diphosphate 4-acetam ...
Escherichia coli
J. Biol. Chem.
240
1305-1308
1965
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2
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