EC Number | Cloned (Comment) | Organism |
---|---|---|
5.1.3.2 | 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 |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
5.1.3.2 | K150R | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type | Mycobacterium tuberculosis |
5.1.3.2 | 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 |
5.1.3.2 | S121A | site-directed mutagenesis, inactive mutant | Mycobacterium tuberculosis |
5.1.3.2 | Y146F | site-directed mutagenesis, inactive mutant | Mycobacterium tuberculosis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
5.1.3.2 | 67000 | - |
about, gel filtration, recombinant enzyme | Mycobacterium tuberculosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.3.2 | UDP-glucose | Mycobacterium tuberculosis | - |
UDP-galactose | - |
r | |
5.1.3.2 | UDP-glucose | Mycobacterium tuberculosis H37Rv | - |
UDP-galactose | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.1.3.2 | Mycobacterium tuberculosis | P9WN67 | - |
- |
5.1.3.2 | Mycobacterium tuberculosis H37Rv | P9WN67 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.1.3.2 | 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 |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
5.1.3.2 | 0.78 | - |
recombinant His-tagged enzyme, pH 8.0, 37°C, substrate UDP-glucose | Mycobacterium tuberculosis |
5.1.3.2 | 7.2 | - |
recombinant His-tagged enzyme, pH 8.0, 37°C, substrate UDP-galactose | Mycobacterium tuberculosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.3.2 | additional information | the type II enzyme also has UDP-GalNAc 4-epimerase activity, EC 5.1.3.7 | Mycobacterium tuberculosis | ? | - |
? | |
5.1.3.2 | additional information | the type II enzyme also has UDP-GalNAc 4-epimerase activity, EC 5.1.3.7 | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
5.1.3.2 | UDP-glucose | - |
Mycobacterium tuberculosis | UDP-galactose | - |
r | |
5.1.3.2 | UDP-glucose | - |
Mycobacterium tuberculosis H37Rv | UDP-galactose | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
5.1.3.2 | dimer | - |
Mycobacterium tuberculosis |
5.1.3.2 | More | enzyme secondary structure analysis using circular dichroism | Mycobacterium tuberculosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.1.3.2 | galE1 | - |
Mycobacterium tuberculosis |
5.1.3.2 | H3634 | - |
Mycobacterium tuberculosis |
5.1.3.2 | Rv3634c | - |
Mycobacterium tuberculosis |
5.1.3.2 | UDP-Gal/Glc 4-epimerase | - |
Mycobacterium tuberculosis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
5.1.3.2 | 37 | - |
assay at | Mycobacterium tuberculosis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.1.3.2 | 8 | - |
assay at | Mycobacterium tuberculosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
5.1.3.2 | NAD+ | associated to enzyme residue Lys150 | Mycobacterium tuberculosis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
5.1.3.2 | 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 |
5.1.3.2 | 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 |
5.1.3.2 | 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 |
5.1.3.2 | 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 |