BRENDA - Enzyme Database

Evidence from 18O feeding studies for hydroxyl group donor in the reaction catalyzed by cytidylate hydroxymethylase MilA

Chen, C.; Gao, T.; Zhao, G.; Deng, Z.; Hu, S.; Xu, H.; He, X.; Chin. Sci. Bull. 58, 864-868 (2013)
No PubMed abstract available

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.2.B2
gene milA, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Streptomyces rimofaciens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
Streptomyces rimofaciens
-
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
?
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
Streptomyces rimofaciens ZJU5119
-
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
?
2.1.2.B2
additional information
Streptomyces rimofaciens
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
?
-
-
-
2.1.2.B2
additional information
Streptomyces rimofaciens ZJU5119
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
?
-
-
-
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
T4virus
the hydroxymethylation is catalyzed by dCMP hydroxymethylase, which transfers a methylene group from methylenetetrahydrofolate to C5 of dCMP and then uses a water molecule to hydrate the methylene group to form hydroxymethyl-dCMP, one precursor for phage DNA replication
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.2.B2
Streptomyces rimofaciens
-
gene milA
-
2.1.2.B2
Streptomyces rimofaciens ZJU5119
-
gene milA
-
2.1.2.8
T4virus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.2.B2
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Streptomyces rimofaciens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
-
735967
Streptomyces rimofaciens
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
-
?
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
-
735967
Streptomyces rimofaciens ZJU5119
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
-
?
2.1.2.B2
additional information
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
735967
Streptomyces rimofaciens
?
-
-
-
-
2.1.2.B2
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
735967
Streptomyces rimofaciens
?
-
-
-
-
2.1.2.B2
additional information
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
735967
Streptomyces rimofaciens ZJU5119
?
-
-
-
-
2.1.2.B2
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
735967
Streptomyces rimofaciens ZJU5119
?
-
-
-
-
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
the hydroxymethylation is catalyzed by dCMP hydroxymethylase, which transfers a methylene group from methylenetetrahydrofolate to C5 of dCMP and then uses a water molecule to hydrate the methylene group to form hydroxymethyl-dCMP, one precursor for phage DNA replication
735967
T4virus
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
-
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
the enzyme uses the solvent water as the hydroxyl group donor
735967
T4virus
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
-
?
2.1.2.8
additional information
5-hydroxymethylcytosine in phage is formed by hydroxymethylation of the cytosine base in deoxycytidylate (dCMP) by deoxycytidylate hydroxymethylase, which uses the solvent water as the hydroxyl group donor
735967
T4virus
?
-
-
-
-
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.2.B2
37
-
assay at
Streptomyces rimofaciens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.2.B2
7.5
-
assay at
Streptomyces rimofaciens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.1.2.B2
5,10-methylenetetrahydrofolate
-
Streptomyces rimofaciens
2.1.2.8
5,10-methylenetetrahydrofolate
-
T4virus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.2.B2
gene milA, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Streptomyces rimofaciens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.2.B2
5,10-methylenetetrahydrofolate
-
Streptomyces rimofaciens
2.1.2.8
5,10-methylenetetrahydrofolate
-
T4virus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
Streptomyces rimofaciens
-
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
?
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
Streptomyces rimofaciens ZJU5119
-
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
?
2.1.2.B2
additional information
Streptomyces rimofaciens
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
?
-
-
-
2.1.2.B2
additional information
Streptomyces rimofaciens ZJU5119
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
?
-
-
-
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
T4virus
the hydroxymethylation is catalyzed by dCMP hydroxymethylase, which transfers a methylene group from methylenetetrahydrofolate to C5 of dCMP and then uses a water molecule to hydrate the methylene group to form hydroxymethyl-dCMP, one precursor for phage DNA replication
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.2.B2
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Streptomyces rimofaciens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
-
735967
Streptomyces rimofaciens
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
-
?
2.1.2.B2
5,10-methylenetetrahydrofolate + H2O + cytidylate
-
735967
Streptomyces rimofaciens ZJU5119
tetrahydrofolate + 5-hydroxymethylcytidylate
-
-
-
?
2.1.2.B2
additional information
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
735967
Streptomyces rimofaciens
?
-
-
-
-
2.1.2.B2
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
735967
Streptomyces rimofaciens
?
-
-
-
-
2.1.2.B2
additional information
5-hydroxymethylcytosine is present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA
735967
Streptomyces rimofaciens ZJU5119
?
-
-
-
-
2.1.2.B2
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
735967
Streptomyces rimofaciens ZJU5119
?
-
-
-
-
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
the hydroxymethylation is catalyzed by dCMP hydroxymethylase, which transfers a methylene group from methylenetetrahydrofolate to C5 of dCMP and then uses a water molecule to hydrate the methylene group to form hydroxymethyl-dCMP, one precursor for phage DNA replication
735967
T4virus
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
-
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
the enzyme uses the solvent water as the hydroxyl group donor
735967
T4virus
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
-
-
?
2.1.2.8
additional information
5-hydroxymethylcytosine in phage is formed by hydroxymethylation of the cytosine base in deoxycytidylate (dCMP) by deoxycytidylate hydroxymethylase, which uses the solvent water as the hydroxyl group donor
735967
T4virus
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.2.B2
37
-
assay at
Streptomyces rimofaciens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.2.B2
7.5
-
assay at
Streptomyces rimofaciens
General Information
EC Number
General Information
Commentary
Organism
2.1.2.B2
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
2.1.2.B2
metabolism
mildiomycin formation pathway catalyzed by MilA and MilB, overview
Streptomyces rimofaciens
2.1.2.B2
additional information
six homologous ORFs originally annotated as putative thymidylate synthase are more likely to be CMP hydroxymethylase
Streptomyces rimofaciens
2.1.2.B2
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
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.1.2.B2
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
2.1.2.B2
metabolism
mildiomycin formation pathway catalyzed by MilA and MilB, overview
Streptomyces rimofaciens
2.1.2.B2
additional information
six homologous ORFs originally annotated as putative thymidylate synthase are more likely to be CMP hydroxymethylase
Streptomyces rimofaciens
2.1.2.B2
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