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Literature summary extracted from

  • Shi, X.; Sha, S.; Liu, L.; Li, X.; Ma, Y.
    A 96-well microtiter plate assay for high-throughput screening of Mycobacterium tuberculosis dTDP-D-glucose 4,6-dehydratase inhibitors (2016), Anal. Biochem., 498, 53-58 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.2.1.46 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

EC Number Application Comment Organism
4.2.1.46 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
4.2.1.46 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)

EC Number Cloned (Comment) Organism
4.2.1.46 expression in Escherichia coli BL21(DE3) Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.46 Ca2+ 0.5 mM, about 50% loss of activity Mycobacterium tuberculosis
4.2.1.46 dTDP competitive inhibitor Mycobacterium tuberculosis
4.2.1.46 dTTP competitive inhibitor Mycobacterium tuberculosis
4.2.1.46 K+ 1 mM, about 10% loss of activity Mycobacterium tuberculosis
4.2.1.46 Mg2+ 0.5 mM, about 30% loss of activity Mycobacterium tuberculosis
4.2.1.46 Mn2+ 0.2 mM, about 50% loss of activity Mycobacterium tuberculosis
4.2.1.46 Na+ 1 mM, about 25% loss of activity Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.46 0.05
-
dTDP-alpha-D-glucose pH 7.5, 30°C Mycobacterium tuberculosis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.46 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.46 39000
-
SDS-PAGE Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.46 Mycobacterium tuberculosis P9WN65
-
-
4.2.1.46 Mycobacterium tuberculosis H37Rv P9WN65
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.46
-
Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.46 dTDP-alpha-D-glucose
-
Mycobacterium tuberculosis dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
?
4.2.1.46 dTDP-alpha-D-glucose
-
Mycobacterium tuberculosis H37Rv dTDP-4-dehydro-6-deoxy-alpha-D-glucose + H2O
-
?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.46 RmlB
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.46 30
-
-
Mycobacterium tuberculosis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.2.1.46 26 60 25°C: about 55% of maximal activity, 60°C: about 40% of maximal activity Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.46 466.6
-
dTDP-alpha-D-glucose pH 7.5, 30°C Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.46 7.5
-
Hepes buffer Mycobacterium tuberculosis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.46 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]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.2.1.46 0.00267
-
dTDP pH 7.5, 30°C Mycobacterium tuberculosis
4.2.1.46 0.00567
-
dTTP pH 7.5, 30°C Mycobacterium tuberculosis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
4.2.1.46 0.006
-
pH 7.5, 30°C Mycobacterium tuberculosis dTDP
4.2.1.46 0.032
-
pH 7.5, 30°C Mycobacterium tuberculosis dTTP

General Information

EC Number General Information Comment Organism
4.2.1.46 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