Any feedback?
Literature summary for 2.3.1.28 extracted from
- Thermoadaptation-directed evolution of chloramphenicol acetyltransferase in an error-prone thermophile using improved procedures (2015), Appl. Microbiol. Biotechnol., 99, 5563-5572.
Application
| Application | Comment | Organism |
|---|---|---|
| molecular biology | the enzyme mutant CATA138T may be useful as a genetic marker in Geobacillus spp. | Staphylococcus aureus |
Cloned(Commentary)
| 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
| 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 [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 [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 25000 | - |
3 * 25000, about, SDS-PAGE | Staphylococcus aureus |
| 75000 | - |
recombinant enzyme, gel filtration | Staphylococcus aureus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acetyl-CoA + chloramphenicol | Staphylococcus aureus | - |
CoA + chloramphenicol 3-acetate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Staphylococcus aureus | - |
- |
- |
Purification (Commentary)
| 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 [micromol/min/mg]
| 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 and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acetyl-CoA + chloramphenicol | - |
Staphylococcus aureus | CoA + chloramphenicol 3-acetate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotrimer | 3 * 25000, about, SDS-PAGE | Staphylococcus aureus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CAT | - |
Staphylococcus aureus |
| chloramphenicol acetyltransferase | - |
Staphylococcus aureus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | 60 | - |
Staphylococcus aureus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 75 | activity range, inactivation at 77°C | Staphylococcus aureus |
Turnover Number [1/s]
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Staphylococcus aureus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| acetyl-CoA | - |
Staphylococcus aureus | |
General Information
| 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 |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
