Cloned (Comment) | Organism |
---|---|
gene thiC, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21AI, which also expresses Azotobacter vinelandii [Fe-S] cluster loading genes (iscSUA, hscBA, fdx, orf3, ndK) | Salmonella enterica |
Protein Variants | Comment | Organism |
---|---|---|
A527T | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
D468N | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
D509G | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
D61N/G513E | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
E281K | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
G273N | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
G355D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
G472D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
G479R | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
G481S | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
G486D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
G92D | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
H501Y | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
additional information | mutant phenotypes, overview | Salmonella enterica |
P498L | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
R397H | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
R544C | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
S247F | random mutagenesis by by hydroxylamine, inactive mutant | Salmonella enterica |
V267M | random mutagenesis by by hydroxylamine, the mutant shows reduced activity compared to the wild-type enzyme | Salmonella enterica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | the enzyme requires an [Fe-S] cluster | Salmonella enterica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | Salmonella enterica | - |
4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate | ? | |
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | Salmonella enterica DM851 | - |
4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Salmonella enterica | - |
serovar Typhimurium LT2, gene thiC | - |
Salmonella enterica DM851 | - |
serovar Typhimurium LT2, gene thiC | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21AI by nickel affinity chromatography and gel filtration | Salmonella enterica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | - |
Salmonella enterica | 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | - |
? | |
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | - |
Salmonella enterica | 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate | ? | |
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | - |
Salmonella enterica DM851 | 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | - |
? | |
5-amino-1-(5-phospho-D-ribosyl)imidazole + S-adenosyl-L-methionine | - |
Salmonella enterica DM851 | 4-amino-2-methyl-5-phosphomethylpyrimidine + 5'-deoxyadenosine + L-methionine + formate + CO | 4-amino-2-methyl-5-phosphomethylpyrimidine is sequentially phosphorylated and combined with 4-methyl-5-(2-hydroxyethyl)-thiazole phosphate to generate thiamine monophosphate before a final phosphorylation generates the active cofactor thiamine diphosphate | ? | |
additional information | reaction assay in the anaerobic chamber | Salmonella enterica | ? | - |
? | |
additional information | reaction assay in the anaerobic chamber | Salmonella enterica DM851 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
HMP-P synthase | - |
Salmonella enterica |
thiC | - |
Salmonella enterica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Salmonella enterica |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
85 | - |
purified recombinant His-tagged wild-type and mutant enzymes, 3 min, inactivation | Salmonella enterica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Salmonella enterica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Salmonella enterica |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the radical S-adenosylmethionine (AdoMet) superfamily | Salmonella enterica |
malfunction | a compromised iron-sulfur ([Fe-S]) cluster metabolism reduces ThiC activity | Salmonella enterica |
metabolism | the enzyme catalyzes the biosynthesis of on part of the thiamine diphosphate cofactor that is essentially used by enzymes in central metabolism such as pyruvate dehydrogenase and2-oxoglutarate dehydrogenase to stabilize the acyl carbanion | Salmonella enterica |