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 | 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 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli | - |
gene lacA | - |
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 | 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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Escherichia coli |
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 | Comment | Organism |
---|---|---|
homotrimer | - |
Escherichia coli |
trimer | 3 * 28000, recombinant enzyme, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
galactoside O-acetyltransferase | - |
Escherichia coli |
galactoside transacetylase | - |
Escherichia coli |
LacA | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
25 | - |
activity assay | Escherichia coli |
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 | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Escherichia coli |