EC Number | Cloned (Comment) | Organism |
---|---|---|
2.1.2.B2 | expressed in Escherichia coli BL21(DE3) cells | Streptomyces rimofaciens |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.1.2.B2 | enzyme complexed with various substrates including CMP, dCMP and 5'-hydroxymethyl-CMP, hanging drop vapor diffusion method, using 15% (w/v) polyethylene glycol 3350, 0.08 M lithium sulfate monohydrate and 0.1 M Tris-HCl, pH 8.5 | Streptomyces rimofaciens |
2.1.2.B2 | structures of MilA and its complexes with substrates including CMP, dCMP and hydroxymethyl-CMP. Two residues in the active site of dCMP hydroxymethylase from T4 phage and thymidylate synthase from Escherichia coli, a serine and an arginine, are respectively replaced by an alanine and a lysine, Ala176 and Lys133, in MilA | Streptomyces rimofaciens |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.1.2.B2 | A176S/K133R | mutation results in a reversal of substrate preference from CMP to dCMP | Streptomyces rimofaciens |
2.1.2.B2 | A176S/K133R | the mutation results in completely eliminated catalytic efficiency toward CMP while its efficiency to dCMP is slightly affected | Streptomyces rimofaciens |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.2.B2 | 0.0719 | - |
CMP | at pH 7.5 and 37°C | Streptomyces rimofaciens | |
2.1.2.B2 | 0.245 | - |
dCMP | at pH 7.5 and 37°C | Streptomyces rimofaciens |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | Streptomyces rimofaciens | - |
tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | Streptomyces rimofaciens ZJU5119 | - |
tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.1.2.B2 | Streptomyces rimofaciens | - |
- |
- |
2.1.2.B2 | Streptomyces rimofaciens | B4Y380 | - |
- |
2.1.2.B2 | Streptomyces rimofaciens ZJU5119 | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.1.2.B2 | Ni-NTA column chromatography and Superdex S200 gel filtration | Streptomyces rimofaciens |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | - |
Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | CMP is the strongly preferred substrate over dCMP | Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | - |
Streptomyces rimofaciens ZJU5119 | tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + CMP | CMP is the strongly preferred substrate over dCMP | Streptomyces rimofaciens ZJU5119 | tetrahydrofolate + 5-hydroxymethyl-CMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + cytidylate | - |
Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethylcytidylate | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + dCMP | - |
Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethyl-dCMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + dCMP | - |
Streptomyces rimofaciens ZJU5119 | tetrahydrofolate + 5-hydroxymethyl-dCMP | - |
? | |
2.1.2.B2 | 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate | - |
Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethyldeoxycytidylate | - |
? | |
2.1.2.B2 | additional information | MilA catalyzes the conversion of CMP into 5-hydroxymethyl-CMP with an efficiency (kcat/KM) of 5fold faster than for deoxycytidylate | Streptomyces rimofaciens | ? | - |
- |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.1.2.B2 | homodimer | - |
Streptomyces rimofaciens |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.1.2.B2 | CMP hydroxymethylase | - |
Streptomyces rimofaciens |
2.1.2.B2 | MilA | - |
Streptomyces rimofaciens |
2.1.2.B2 | thymidylate synthase | - |
Streptomyces rimofaciens |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.2.B2 | 0.032 | - |
dCMP | at pH 7.5 and 37°C | Streptomyces rimofaciens | |
2.1.2.B2 | 0.047 | - |
CMP | at pH 7.5 and 37°C | Streptomyces rimofaciens |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.1.2.B2 | metabolism | the enzyme is involved in mildiomycin biosynthesis | Streptomyces rimofaciens |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.2.B2 | 0.13 | - |
dCMP | at pH 7.5 and 37°C | Streptomyces rimofaciens | |
2.1.2.B2 | 0.65 | - |
CMP | at pH 7.5 and 37°C | Streptomyces rimofaciens |