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Literature summary for 2.3.1.18 extracted from
- Tyrosine 83 is essential for the activity of E. coli galactoside transacetylase (2007), Biochim. Biophys. Acta, 1774, 243-248.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene lacA, overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli TOP10 cells | Escherichia coli |
| into the pTrcHisB vector | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| Y83F | site-directed mutagenesis, the mutant shows highy reduced activity compared to the wild-type enzyme | Escherichia coli |
| Y83F | tyrosine83 is essential for the activity of galactoside transacetylase | Escherichia coli |
| Y83L | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
| Y83L | tyrosine83 is essential for the activity of galactoside transacetylase | Escherichia coli |
| Y83V | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
| Y83V | tyrosine83 is essential for the activity of galactoside transacetylase | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 12 | - |
p-nitrophenyl-beta-D-galactopyranoside | H155A variant | Escherichia coli |  |
| 24.1 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83L | Escherichia coli | |
| 24.1 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83L mutant | Escherichia coli | |
| 27.2 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, wild-type enzyme | Escherichia coli | |
| 27.2 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged wild type enzyme | Escherichia coli | |
| 29.5 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83F | Escherichia coli | |
| 29.5 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83F mutant | Escherichia coli | |
| 60 | - |
p-nitrophenyl-beta-D-galactopyranoside | wild type enzyme | Escherichia coli | |
| 97 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83V | Escherichia coli | |
| 97 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83V mutant | Escherichia coli | |
| 629 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged wild type enzyme | Escherichia coli | |
| 770 | - |
isopropyl-thio-beta-D-galactopyranoside | wild type enzyme | Escherichia coli | |
| 1150 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83V mutant | Escherichia coli | |
| 2440 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83L mutant | Escherichia coli | |
| 2720 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83F mutant | Escherichia coli | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 28000 | - |
determined by SDS-PAGE and gel filtration, monomer, containing a hexahistidine tag | Escherichia coli |
| 80000 | - |
recombinant enzyme, gel filtration | Escherichia coli |
| 80000 | - |
determined by SDS-PAGE and gel filtration, trimer, containing hexahistidine tags | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acetyl-CoA + a beta-D-galactoside | Escherichia coli | - |
CoA + a 6-acetyl-beta-D-galactoside | - |
? | |
| acetyl-CoA + beta-D-galactoside | Escherichia coli | - |
CoA + 6-acetyl-beta-D-galactoside | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Escherichia coli | - |
gene lacA | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged wild-type and mutant enzymes by nickel affinity chromatography, ion exchange chromatography, and gel filtration | Escherichia coli |
| using a HiTrap metal-chelating affinity chromatography column, a HiTrap desalting, and a Superdex 200 column | Escherichia coli |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| acetyl-CoA + a beta-D-galactoside = CoA + a 6-acetyl-beta-D-galactoside | Tyr83 is essential for enzyme activity | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl beta-D-galactopyranoside + acetyl-CoA | - |
Escherichia coli | 4-nitrophenyl 6-O-acetyl-beta-D-galactopyranoside + CoA | - |
? | |
| acetyl-CoA + a beta-D-galactoside | - |
Escherichia coli | CoA + a 6-acetyl-beta-D-galactoside | - |
? | |
| acetyl-CoA + beta-D-galactoside | - |
Escherichia coli | CoA + 6-acetyl-beta-D-galactoside | - |
? | |
| isopropyl-thio-beta-D-galactopyranoside | - |
Escherichia coli | ? | - |
? | |
| isopropyl-thio-beta-D-galactopyranoside + acetyl-CoA | - |
Escherichia coli | 6-acetyl-isopropyl-thio-beta-D-galactopyranoside + CoA | - |
? | |
| isopropyl-thio-beta-D-galactopyranoside + acetyl-CoA | - |
Escherichia coli | isopropyl 6-O-acetyl-1-thio-beta-D-galactopyranoside + CoA | - |
? | |
| p-nitrophenyl-beta-D-galactopyranoside | - |
Escherichia coli | ? | - |
? | |
| p-nitrophenyl-beta-D-galactopyranoside + acetyl-CoA | - |
Escherichia coli | p-nitrophenyl 6-O-acetyl-beta-D-galactopyranoside + CoA | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotrimer | - |
Escherichia coli |
| trimer | 3 * 28000, recombinant enzyme, SDS-PAGE | Escherichia coli |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| galactoside O-acetyltransferase | - |
Escherichia coli |
| galactoside transacetylase | - |
Escherichia coli |
| LacA | - |
Escherichia coli |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Escherichia coli |
| 25 | - |
activity assay | Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.006 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83L | Escherichia coli | |
| 0.006 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83L mutant | Escherichia coli | |
| 0.0082 | - |
p-nitrophenyl-beta-D-galactopyranoside | H155A variant | Escherichia coli | |
| 0.022 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83V | Escherichia coli | |
| 0.022 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83V mutant | Escherichia coli | |
| 0.175 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83V mutant | Escherichia coli | |
| 0.31 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83L mutant | Escherichia coli | |
| 4.9 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, mutant Y83F | Escherichia coli | |
| 4.9 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged Y83F mutant | Escherichia coli | |
| 11.1 | - |
4-nitrophenyl-beta-D-galactopyranoside | 25°C, wild-type enzyme | Escherichia coli | |
| 11.1 | - |
p-nitrophenyl-beta-D-galactopyranoside | hexahistidine-tagged wild type enzyme | Escherichia coli | |
| 14.7 | - |
p-nitrophenyl-beta-D-galactopyranoside | wild type enzyme | Escherichia coli | |
| 122 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged wild type enzyme | Escherichia coli | |
| 133 | - |
isopropyl-thio-beta-D-galactopyranoside | wild type enzyme | Escherichia coli | |
| 168 | - |
isopropyl-thio-beta-D-galactopyranoside | hexahistidine-tagged Y83F mutant | Escherichia coli | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| acetyl-CoA | - |
Escherichia coli | |
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