Application | Comment | Organism |
---|---|---|
molecular biology | the enzyme mutant CATA138T may be useful as a genetic marker in Geobacillus spp. | Staphylococcus aureus |
Cloned (Comment) | Organism |
---|---|
gene cat, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-CodonPlus(DE3)-RIPL and in Geobacillus kaustophilus strain MK480 | Staphylococcus aureus |
Protein Variants | Comment | Organism |
---|---|---|
A138S | site-directed mutagenesis, the enzyme mutant shows increased thermostability at 60-65°C for 24 h compared to the wild-type enzyme, thermostability enhancement results from the A138T replacement and can attributed to both the presence of a hydroxyl group and the bulk of the threonine side chain. CAT A138S mutation confers chloramphenicol resistance to Geobacillus kaustophilus cells at high temperature more efficiently than the wild-type enzyme | Staphylococcus aureus |
A138T | site-directed mutagenesis, the enzyme mutant shows increased thermostability at 60-65°C for 24 h compared to the wild-type enzyme, thermostability enhancement results from the A138T replacement and can attributed to both the presence of a hydroxyl group and the bulk of the threonine side chain. CAT A138T mutation confers chloramphenicol resistance to Geobacillus kaustophilus cells at high temperature more efficiently than the wild-type enzyme. The A138T substitution has no effect on CAT activity | Staphylococcus aureus |
A138V | site-directed mutagenesis, the enzyme mutant shows highly increased thermostability at 60-65°C for 24 h compared to the wild-type enzyme, thermostability enhancement results from the A138T replacement and can attributed to both the presence of a hydroxyl group and the bulk of the threonine side chain. CAT A138V mutation confers chloramphenicol resistance to Geobacillus kaustophilus cells at high temperature more efficiently than the wild-type enzyme | Staphylococcus aureus |
additional information | thermoadaptation-directed enzyme evolution approach for generation of mutant genes encoding enzyme variants that are more thermostable than the parent enzyme using an error-prone thermophile strain MK480 derived from Geobacillus kaustophilus strain HTA426, increase of the thermostability of the chloramphenicol acetyltransferase (CAT) from Staphylococcus aureus and successfully generation of a CAT variant with an A138 replacement (CATA138X), method, overview | Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.153 | - |
chloramphenicol | pH 7.0, 60°C, recombinant mutant A138T | Staphylococcus aureus | |
0.162 | - |
chloramphenicol | pH 7.0, 60°C, recombinant wild-type enzyme | Staphylococcus aureus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
25000 | - |
3 * 25000, about, SDS-PAGE | Staphylococcus aureus |
75000 | - |
recombinant enzyme, gel filtration | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + chloramphenicol | Staphylococcus aureus | - |
CoA + chloramphenicol 3-acetate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and gel filtration | Staphylococcus aureus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
140 | - |
pH 7.0, 60°C, purified recombinant mutant A138T | Staphylococcus aureus |
240 | - |
pH 7.0, 60°C, purified recombinant mutant A138V | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + chloramphenicol | - |
Staphylococcus aureus | CoA + chloramphenicol 3-acetate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotrimer | 3 * 25000, about, SDS-PAGE | Staphylococcus aureus |
Synonyms | Comment | Organism |
---|---|---|
CAT | - |
Staphylococcus aureus |
chloramphenicol acetyltransferase | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | 60 | - |
Staphylococcus aureus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 75 | activity range, inactivation at 77°C | Staphylococcus aureus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
193 | - |
chloramphenicol | pH 7.0, 60°C, recombinant mutant A138T | Staphylococcus aureus | |
209 | - |
chloramphenicol | pH 7.0, 60°C, recombinant wild-type enzyme | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Staphylococcus aureus |
General Information | Comment | Organism |
---|---|---|
evolution | CAT members are currently classified into three types on the basis of their substrate specificities. Staphylococcus aureus CAT is classified as type III | Staphylococcus aureus |
additional information | homology modeling of enzyme CAT structures using the crystal structure of Escherichia coli CAT, PDB ID 3CLA, overview. Residue A138 resides on beta-sheet 7 and is located on the protein surfaces far from the active center. Although the side chains of I134, P135, and L164 surround this residue, no hydrophobic interactions are observed within 4 A. Modeling of CATA138T, overview | Staphylococcus aureus |