Cloned (Comment) | Organism |
---|---|
gene milA, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptomyces rimofaciens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + H2O + cytidylate | Streptomyces rimofaciens | - |
tetrahydrofolate + 5-hydroxymethylcytidylate | - |
? | |
5,10-methylenetetrahydrofolate + H2O + cytidylate | Streptomyces rimofaciens ZJU5119 | - |
tetrahydrofolate + 5-hydroxymethylcytidylate | - |
? | |
additional information | Streptomyces rimofaciens | 5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA | ? | - |
? | |
additional information | Streptomyces rimofaciens ZJU5119 | 5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces rimofaciens | - |
gene milA | - |
Streptomyces rimofaciens ZJU5119 | - |
gene milA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Streptomyces rimofaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + H2O + cytidylate | - |
Streptomyces rimofaciens | tetrahydrofolate + 5-hydroxymethylcytidylate | - |
? | |
5,10-methylenetetrahydrofolate + H2O + cytidylate | - |
Streptomyces rimofaciens ZJU5119 | tetrahydrofolate + 5-hydroxymethylcytidylate | - |
? | |
additional information | 5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA | Streptomyces rimofaciens | ? | - |
? | |
additional information | MilA uses solvent water as the hydroxyl group donor like cytidylate hydroxymethylase. Thr102 of MilA is a potential critical amino acid anchoring one molecule of water for hydroxylation. Thr102 of MilA is predicted as potential critical amino acid anchoring one molecule of water for hydroxylation | Streptomyces rimofaciens | ? | - |
? | |
additional information | 5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA | Streptomyces rimofaciens ZJU5119 | ? | - |
? | |
additional information | MilA uses solvent water as the hydroxyl group donor like cytidylate hydroxymethylase. Thr102 of MilA is a potential critical amino acid anchoring one molecule of water for hydroxylation. Thr102 of MilA is predicted as potential critical amino acid anchoring one molecule of water for hydroxylation | Streptomyces rimofaciens ZJU5119 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cytidylate hydroxymethylase | - |
Streptomyces rimofaciens |
MilA | - |
Streptomyces rimofaciens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptomyces rimofaciens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Streptomyces rimofaciens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
5,10-methylenetetrahydrofolate | - |
Streptomyces rimofaciens |
General Information | Comment | Organism |
---|---|---|
evolution | since MilA have similar mechanism as CH in hydroxyl group formation, it may belong to the superfamily of thymidylate synthase and cytidylate hydroxymethylase | Streptomyces rimofaciens |
metabolism | mildiomycin formation pathway catalyzed by MilA and MilB, overview | Streptomyces rimofaciens |
additional information | six homologous ORFs originally annotated as putative thymidylate synthase are more likely to be CMP hydroxymethylase | Streptomyces rimofaciens |
physiological function | several nucleotide-derived nucleoside antibiotics feature modified bases such as hydroxymethyl pyrimidine in polyoxin, and hydroxymethyl cytosine in mildiomycin. The CMP hydroxymethylase named MilA in the biosynthetic pathway of nucleoside mildiomycin in Streptoverticillum rimofaciens ZJU5119 can convert CMP in vitro in the presence of tetrahydrofolate to 5-hydroxymethyl-CMP that is immediately hydrolyzed into free 5-hydroxymethylcytosine by MilB | Streptomyces rimofaciens |