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Literature summary for 5.1.3.2 extracted from

  • Pardeshi, P.; Rao, K.K.; Balaji, P.V.
    Rv3634c from Mycobacterium tuberculosis H37Rv encodes an enzyme with UDP-Gal/Glc and UDP-GalNAc 4-epimerase activities (2017), PLoS ONE, 12, e0175193 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.3.2

Cloned(Commentary)

Cloned (Comment) Organism
gene galE1 or Rv3634c, sequence comparisons, e.g. with the enzyme from Mycobacterium smegmatis strain mc2 4517, recombinant expression of His-tagged enzyme in Escherichia coli strains BL21(DE3)pLysS, BL21(DE3), BL21-CodonPlus(DE3)-RIPL, OverExpress C41(DE3), or OverExpress C43(DE3)pLysS, and recombinant expression of the enzyme in Mycobacterium smegmatis strain mc2 4517, none of these approaches leads to expression of the enzyme in the soluble form, except for strain BL21(DE3)pLysS, recombinant expression of enzyme mutants Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
K150R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Mycobacterium tuberculosis
additional information addition of NAD+ to Lys150 significantly abrogates the loss of activity. None of the mutations affected the quaternary structure of the protein Mycobacterium tuberculosis
S121A site-directed mutagenesis, inactive mutant Mycobacterium tuberculosis
Y146F site-directed mutagenesis, inactive mutant Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
67000
-
 about, gel filtration, recombinant enzyme Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-glucose Mycobacterium tuberculosis
-
 UDP-galactose
-
 r
UDP-glucose Mycobacterium tuberculosis H37Rv
-
 UDP-galactose
-
 r

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WN67
-
-
Mycobacterium tuberculosis H37Rv P9WN67
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, ultrafiltration, and gel filtration Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.78
-
 recombinant His-tagged enzyme, pH 8.0, 37°C, substrate UDP-glucose Mycobacterium tuberculosis
7.2
-
 recombinant His-tagged enzyme, pH 8.0, 37°C, substrate UDP-galactose Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the type II enzyme also has UDP-GalNAc 4-epimerase activity, EC 5.1.3.7 Mycobacterium tuberculosis ?
-
 ?
additional information the type II enzyme also has UDP-GalNAc 4-epimerase activity, EC 5.1.3.7 Mycobacterium tuberculosis H37Rv ?
-
 ?
UDP-glucose
-
 Mycobacterium tuberculosis UDP-galactose
-
 r
UDP-glucose
-
 Mycobacterium tuberculosis H37Rv UDP-galactose
-
 r

Subunits

Subunits Comment Organism
dimer
-
 Mycobacterium tuberculosis
More enzyme secondary structure analysis using circular dichroism Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
galE1
-
 Mycobacterium tuberculosis
H3634
-
 Mycobacterium tuberculosis
Rv3634c
-
 Mycobacterium tuberculosis
UDP-Gal/Glc 4-epimerase
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
NAD+ associated to enzyme residue Lys150 Mycobacterium tuberculosis

General Information

General Information Comment Organism
evolution sugar nucleotide epimerases, including UDP-Gal/Glc 4-epimerase, are a class of enzymes that belong to the extended SDR subfamily of the short chain dehydrogenase reductase (SDR) superfamily, enzyme Rv3634c belongs to the NAD+-dependent epimerase/dehydratase family of proteins (accession no. PF01370) Mycobacterium tuberculosis
malfunction mutations S121A and Y146F lead to complete loss of activity whereas mutation K150R leads to partial loss of activity. There are no gross changes in the secondary structures of any of these three mutants Mycobacterium tuberculosis
additional information Ser121 and Tyr146 are essential for epimerase activity of Rv3634c. The catalytic triad consisting of Ser, Tyr and Lys carries out proton transfer from nucleotide sugar to NAD+ and back, thus effecting the epimerization of the substrate Mycobacterium tuberculosis
physiological function UDP-Gal/GalNAc epimerases differ from each other in their substrate specificity with respect to the utilization of N-acetylated derivatives and are accordingly classified into three types viz., type I, type II and type III. Type I enzymes utilize only UDP-Glc/Gal as substrates whereas Type III enzymes use only UDP-GlcNAc/GalNAc as substrates. Type II enzymes can utilize both as substrates Mycobacterium tuberculosis