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Literature summary for 2.3.1.157 extracted from

  • Zhang, Z.; Shimizu, Y.; Kawarabayasi, Y.
    Characterization of the amino acid residues mediating the unique amino-sugar-1-phosphate acetyltransferase activity of the archaeal ST0452 protein (2015), Extremophiles, 19, 417-427.
    View publication on PubMed

Application

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(Commentary)

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

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

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 [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

Metals/Ions Comment Organism Structure
Mg2+ required Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Sulfurisphaera tokodaii Q975F9
-
-
Sulfurisphaera tokodaii 7 Q975F9
-
-
Sulfurisphaera tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7 Q975F9
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Sulfurisphaera tokodaii

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
More the enzyme contains the the N-terminal nucleotidylyltransferase domain (residues 1–210) and the C-terminal acetyltransferase domain (residues 211–401), 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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
assay at Sulfurisphaera tokodaii

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Sulfurisphaera tokodaii

Cofactor

Cofactor Comment Organism Structure
acetyl-CoA
-
Sulfurisphaera tokodaii

General Information

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 [mM/s]

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