Application | Comment | Organism |
---|---|---|
additional information | the increase in activity induced by some substitutions and truncations may be a useful feature that can be exploited for commercial application of this enzyme | Sulfurisphaera tokodaii |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain BL21-Codon Plus(DE3)-RIL | Sulfurisphaera tokodaii |
gene ST0452, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Codon Plus(DE3)-RIL | Sulfurisphaera tokodaii |
Protein Variants | Comment | Organism |
---|---|---|
H308A | specific activity is 0.7% compared to the wild-type enzyme | Sulfurisphaera tokodaii |
H308A | site-directed mutagenesis, the mutation diminishes both amino-sugar-1-P AcTase activities of the ST0452 protein. The mutant shows 7.7% and 0.7% of wild-type GalN-1-P AcTase and GlcN-1-P AcTase activity, respetively | Sulfurisphaera tokodaii |
K337A | site-directed mutagenesis, the mutant enzyme shows slightly decreasing GalN-1-P AcTase activity and slightly increasing GlcN-1-P AcTase activity compared to the wild-type enzyme. The mutant shows 82.6% and 137.7% of wild-type GalN-1-P AcTase and GlcN-1-P AcTase activity, respetively | Sulfurisphaera tokodaii |
K340A | specific activity is 147.1% compared to the wild-type enzyme | Sulfurisphaera tokodaii |
K340A | site-directed mutagenesis, the mutant enzyme shows moderately decreasing GalN-1-P AcTase activity and moderately increasing GlcN-1-P AcTase activity compared to the wild-type enzyme. The mutant shows 63.3% and 147.1% of wild-type GalN-1-P AcTase and GlcN-1-P AcTase activity, respetively | Sulfurisphaera tokodaii |
K377A | specific activity is 137.7% compared to the wild-type enzyme | Sulfurisphaera tokodaii |
additional information | glucosamine-1-phosphate acetyltransferase activity of C-terminal deletion mutants DC005 and DC011 (deletion of the C-terminal 5 or 11 residues of the ST0452 protein) are respectively, 4.8 and 16.8 times higher than that of the wild-type ST0452 protein. The mutant enzyme DC011 (deletion of the C-terminal 11 residues of the ST0452 protein) shows little thermal stability at 80°C. The C-terminal domain of the ST0452 protein, with its LbetaH structure, appears to be essential for the formation of its trimeric form and, in turn, the high stability of the entire ST0452 protein. The deletion mutant enzymes DC021, DC031, DC041, DC071 and DC121, are produced in an insoluble form or aggregated immediately after purification. Mutant enzymes DC051 and DC171 can be expressed in a soluble form. Mutant enzyme DC051 becomes completely insoluble after 5 min treatment at 60°C, while mutant enzyme DC171 is insoluble after 5 min treatment at 70 °C | Sulfurisphaera tokodaii |
additional information | construction of expression vectors encoding a series of ST0452 C-terminal deletion mutants with hexahistidine tags at their C-termini, designated pST0452(DC005)H, pST0452(DC011)H, pST0452(DC021)H, pST0452(DC031)H, pST0452(DC041) H, pST0452(DC051)H, pST0452(DC071)H, pST0452 (DC121)H and pST0452(DC171)H. The deletion mutants retain the same tertiary structures as the wild-type ST0452 protein, but some show an altered thermostability, overview | Sulfurisphaera tokodaii |
N331A | specific activity is 46.1% compared to the wild-type enzyme | Sulfurisphaera tokodaii |
N331A | site-directed mutagenesis, the mutant enzyme shows highly decreasing GalN-1-P AcTase activity and decreasing GlcN-1-P AcTase activity compared to the wild-type enzyme. The mutant shows 3.1% and 46.1% of wild-type GalN-1-P AcTase and GlcN-1-P AcTase activity, respetively | Sulfurisphaera tokodaii |
Y311A | specific activity is 118.4% compared to the wild-type enzyme | Sulfurisphaera tokodaii |
Y311A | site-directed mutagenesis, the mutant enzyme shows highly decreasing GalN-1-P AcTase activity and increasing GlcN-1-P AcTase activity compared to the wild-type enzyme. The mutant shows 3.3% and 118.4% of wild-type GalN-1-P AcTase and GlcN-1-P AcTase activity, respetively | Sulfurisphaera tokodaii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | residues Tyr311, Lys337 and Lys340 plus C-terminal 11-residue region of the ST0452 protein enhance its GalN-1-P AcTase activity and suppress its GlcN-1-P AcTase activity, this function might be lost in bacterial enzymes | Sulfurisphaera tokodaii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.52 | - |
acetyl-CoA | pH 7.5, 80°C, mutant enzyme DC005 with a deletion of the C-terminal 5 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
0.52 | - |
acetyl-CoA | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
0.56 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC005 with a deletion of the C-terminal 5 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
0.56 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
0.59 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.59 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
0.63 | - |
acetyl-CoA | pH 7.5, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.63 | - |
acetyl-CoA | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
0.66 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii | |
0.84 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
1.55 | - |
acetyl-CoA | pH 7.5, 80°C, mutant enzyme DC011 with a deletion of the C-terminal 11 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
1.55 | - |
acetyl-CoA | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii | |
1.69 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC011 with a deletion of the C-terminal 11 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
1.69 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii | |
1.71 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Sulfurisphaera tokodaii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + alpha-D-galactosamine 1-phosphate | Sulfurisphaera tokodaii | - |
CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-galactosamine 1-phosphate | Sulfurisphaera tokodaii 7 | - |
CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-galactosamine 1-phosphate | Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | - |
CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | Sulfurisphaera tokodaii | - |
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | Sulfurisphaera tokodaii 7 | - |
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | - |
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sulfurisphaera tokodaii | Q975F9 | - |
- |
Sulfurisphaera tokodaii 7 | Q975F9 | - |
- |
Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | Q975F9 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Sulfurisphaera tokodaii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.35 | - |
pH 7.5, 80°C, mutant enzyme H308A | Sulfurisphaera tokodaii |
3 | 8 | alpha-D-galactosamine 1-phosphate substrate, pH 7.5, 80°C, recombinant mutant D005 protein | Sulfurisphaera tokodaii |
23.1 | - |
pH 7.5, 80°C, mutant enzyme N331A | Sulfurisphaera tokodaii |
29.6 | - |
alpha-D-galactosamine 1-phosphate aubstrate, pH 7.5, 80°C, recombinant mutant D011 protein | Sulfurisphaera tokodaii |
47.6 | - |
alpha-D-galactosamine 1-phosphate aubstrate, pH 7.5, 80°C, recombinant wild-type ST0452 protein | Sulfurisphaera tokodaii |
50 | - |
pH 7.5, 80°C, wild-type enzyme enzyme | Sulfurisphaera tokodaii |
59.2 | - |
pH 7.5, 80°C, mutant enzyme Y311A | Sulfurisphaera tokodaii |
67.4 | - |
alpha-D-glucosamine 1-phosphate substrate, pH 7.5, 80°C, recombinant wild-type ST0452 protein | Sulfurisphaera tokodaii |
68.9 | - |
pH 7.5, 80°C, mutant enzyme K337A | Sulfurisphaera tokodaii |
73.5 | - |
pH 7.5, 80°C, mutant enzyme K340A | Sulfurisphaera tokodaii |
323.5 | - |
alpha-D-glucosamine 1-phosphate substrate, pH 7.5, 80°C, recombinant mutant D005 protein | Sulfurisphaera tokodaii |
1131 | - |
alpha-D-glucosamine 1-phosphate aubstrate, pH 7.5, 80°C, recombinant mutant D011 protein | Sulfurisphaera tokodaii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + alpha-D-galactosamine 1-phosphate | - |
Sulfurisphaera tokodaii | CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-galactosamine 1-phosphate | - |
Sulfurisphaera tokodaii 7 | CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-galactosamine 1-phosphate | - |
Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | CoA + N-acetyl-alpha-D-galactosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | - |
Sulfurisphaera tokodaii | CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | - |
Sulfurisphaera tokodaii 7 | CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? | |
acetyl-CoA + alpha-D-glucosamine 1-phosphate | - |
Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | CoA + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
? | |
additional information | the enzyme has multiple sugar-1-phosphate nucleotidylyltransferase, EC 2.7.7.37, and amino-sugar-1-phosphate acetyltransferase, EC 2.3.1.157, activities, overview. In addition to glucosamine-1-phosphate acetyltransferase activity, it possesses unique galactosamine-1-phosphate acetyltransferase activity. Also, the enzyme possesses GlcNAc-1-phosphate nucleotidylyltransferase, EC 2.7.7.23, and N-acetyl-D-galactosamine-1-phosphate uridyltransferase, EC 2.7.7.83, activities, as well as the expected glucose-1-phosphate thymidylyltransferase, EC 2.7.7.24, activity. The ST0452 protein can catalyze the acetylation of both GlcN-1-P and GalN-1-P, while GalN-1-P AcTase activity is not detected in bacterial enzymes | Sulfurisphaera tokodaii | ? | - |
? | |
additional information | the enzyme has multiple sugar-1-phosphate nucleotidylyltransferase, EC 2.7.7.37, and amino-sugar-1-phosphate acetyltransferase, EC 2.3.1.157, activities, overview. In addition to glucosamine-1-phosphate acetyltransferase activity, it possesses unique galactosamine-1-phosphate acetyltransferase activity. Also, the enzyme possesses GlcNAc-1-phosphate nucleotidylyltransferase, EC 2.7.7.23, and N-acetyl-D-galactosamine-1-phosphate uridyltransferase, EC 2.7.7.83, activities, as well as the expected glucose-1-phosphate thymidylyltransferase, EC 2.7.7.24, activity. The ST0452 protein can catalyze the acetylation of both GlcN-1-P and GalN-1-P, while GalN-1-P AcTase activity is not detected in bacterial enzymes | Sulfurisphaera tokodaii 7 | ? | - |
? | |
additional information | the enzyme has multiple sugar-1-phosphate nucleotidylyltransferase, EC 2.7.7.37, and amino-sugar-1-phosphate acetyltransferase, EC 2.3.1.157, activities, overview. In addition to glucosamine-1-phosphate acetyltransferase activity, it possesses unique galactosamine-1-phosphate acetyltransferase activity. Also, the enzyme possesses GlcNAc-1-phosphate nucleotidylyltransferase, EC 2.7.7.23, and N-acetyl-D-galactosamine-1-phosphate uridyltransferase, EC 2.7.7.83, activities, as well as the expected glucose-1-phosphate thymidylyltransferase, EC 2.7.7.24, activity. The ST0452 protein can catalyze the acetylation of both GlcN-1-P and GalN-1-P, while GalN-1-P AcTase activity is not detected in bacterial enzymes | Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme contains the the N-terminal nucleotidylyltransferase domain (residues 1210) and the C-terminal acetyltransferase domain (residues 211401), respectively. Comparisons of the crystal structures of the ST0452 protein, PDB ID GGO, and Escherichia coli protein EcGlmU2, PDB ID 2OI5, comparison with ST0452 mutant enzymes, overview. Despite the structural similarities between the N- and C-termini of the ST0452 protein and those of Escherichia coli EcGlmU, the thermostabilities of the two proteins differ greatly, as EcGlmU is a mesophilic enzyme. The structures of these proteins do not correlate directly with their thermostability. The distance between the GlcN-1-P AcTase and GlcNAc-1-P UTase catalytic centers is smaller in the ST0452 protein than the mesophilic bacterial GlmU | Sulfurisphaera tokodaii |
trimer | the C-terminal domain of the ST0452 protein, with its LbetaH structure, appears to be essential for the formation of its trimeric form and, in turn, the high stability of the entire ST0452 protein | Sulfurisphaera tokodaii |
Synonyms | Comment | Organism |
---|---|---|
amino-sugar-1-P AcTase | - |
Sulfurisphaera tokodaii |
amino-sugar-1-phosphate acetyltransferase | - |
Sulfurisphaera tokodaii |
galactosamine-1-phosphate acetyltransferase | - |
Sulfurisphaera tokodaii |
GalN-1-P AcTase | - |
Sulfurisphaera tokodaii |
GlcN-1-P AcTase | - |
Sulfurisphaera tokodaii |
glucosamine-1-phosphate acetyltransferase | - |
Sulfurisphaera tokodaii |
ST0452 | - |
Sulfurisphaera tokodaii |
ST0452 protein | - |
Sulfurisphaera tokodaii |
STK_04520 | locus name | Sulfurisphaera tokodaii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
assay at | Sulfurisphaera tokodaii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
mutant enzyme DC005 shows the same thermostability as wild-type ST0452 protein, whereas mutant enzyme DC011 denatures and becomes insoluble by 5-min treatment at 80 °C. The C-terminal domain of the ST0452 protein, with its LbetaH structure, appears to be essential for the formation of its trimeric form and, in turn, the high stability of the entire ST0452 protein | Sulfurisphaera tokodaii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12.6 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii | |
17.9 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
69.7 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
123.2 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
123.2 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
490.6 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC005 with a deletion of the C-terminal 5 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
490.6 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
2311 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC011 with a deletion of the C-terminal 11 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
2311 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Sulfurisphaera tokodaii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Sulfurisphaera tokodaii |
General Information | Comment | Organism |
---|---|---|
additional information | the C-terminal tail region of the ST0452 protein might be important for recognition of the multiple substrates for amino-sugar-1-P AcTase activity. The ST0452 protein contains only two Cys residues, it is unlikely that Cys-Cys bonds contribute to its thermostability. Residue Asn331 in the ST0452 protein is essential for the GalN-1-P AcTase activity, but it is much less important and not essential for the GlcN-1-P AcTase activity. The C-terminal residues of the ST0452 protein enhance the turnover rate of its GalN-1-P AcTase catalytic activity and slightly suppress substrate binding. Residue H308 is essential for both amino-sugar-1-P AcTase activities of the ST0452 protein | Sulfurisphaera tokodaii |
physiological function | the archaeal enzyme's high GalN-1-P AcTase activity, which is not detected on the bacterial and eukaryotic similar enzymes, supports the production of the UDP-GalNAc in archaeal cells | Sulfurisphaera tokodaii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
19.6 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii | |
21.5 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
40.8 | - |
alpha-D-galactosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
211 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
211 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, wild-type ST0452 protein | Sulfurisphaera tokodaii | |
869.2 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC005 with a deletion of the C-terminal 5 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
869.2 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D005 of ST0452 protein | Sulfurisphaera tokodaii | |
1489 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, mutant enzyme DC011 with a deletion of the C-terminal 11 residues of the ST0452 protein | Sulfurisphaera tokodaii | |
1489 | - |
alpha-D-glucosamine 1-phosphate | pH 7.5, 80°C, deletion mutant D011 of ST0452 protein | Sulfurisphaera tokodaii |