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Literature summary for 3.2.1.54 extracted from
- Recombinant expression, purification, and characterization of a cyclodextrinase from Massilia timonae (2019), Protein Expr. Purif., 154, 74-84 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21 Star(DE3) cells | Massilia timonae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| CTAB | complete inhibition at 5 mM | Massilia timonae |  |
| Cu2+ | 3.6% residual activity at 5 mM | Massilia timonae | |
| EDTA | complete inhibition at 5 mM | Massilia timonae | |
| Fe3+ | 13.9% residual activity at 5 mM | Massilia timonae | |
| K+ | 78% residual activity at 10 mM | Massilia timonae | |
| Mg2+ | 60.7% residual activity at 5 mM, complete inhibition at 10 mM | Massilia timonae | |
| Mn2+ | 76.2% residual activity at 5 mM, complete inhibition at 10 mM | Massilia timonae | |
| Na+ | 65.9% residual activity at 10 mM | Massilia timonae | |
| SDS | complete inhibition at 5 mM | Massilia timonae | |
| Tween 80 | 62.1% residual activity at 1.0% (v/v) | Massilia timonae | |
| Zn2+ | complete inhibition at 5 mM | Massilia timonae | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2.1 | - |
beta-cyclodextrin | at pH 7.5 and 40°C | Massilia timonae | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | 276.8% activity at 5 mM | Massilia timonae | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 260000 | - |
gel filtration | Massilia timonae |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Massilia timonae | K9DE84 | - |
- |
| Massilia timonae CCUG 45783 | K9DE84 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| HisTrap nickel column chromatography | Massilia timonae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| beta-cyclodextrin + H2O | 100% activity | Massilia timonae | D-glucose + maltose | - |
? | |
| maltodextrin + H2O | 62% activity compared to beta-cyclodextrin | Massilia timonae | ? | - |
? | |
| maltodextrin + H2O | 62% activity compared to beta-cyclodextrin | Massilia timonae CCUG 45783 | ? | - |
? | |
| maltoheptaose + H2O | - |
Massilia timonae | D-glucose + maltose + ? | - |
? | |
| maltoheptaose + H2O | - |
Massilia timonae CCUG 45783 | D-glucose + maltose + ? | - |
? | |
| maltotetraose + H2O | - |
Massilia timonae | 2 D-glucose + maltose | - |
? | |
| maltotetraose + H2O | - |
Massilia timonae CCUG 45783 | 2 D-glucose + maltose | - |
? | |
| additional information | no activity with maltose | Massilia timonae | ? | - |
? | |
| additional information | no activity with maltose | Massilia timonae CCUG 45783 | ? | - |
? | |
| starch + H2O | 8% activity compared to beta-cyclodextrin | Massilia timonae | ? | - |
? | |
| starch + H2O | 8% activity compared to beta-cyclodextrin | Massilia timonae CCUG 45783 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotetramer | 4 * 65000, SDS-PAGE | Massilia timonae |
| homotetramer | 4 * 69400, calculated from amino acid sequence | Massilia timonae |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| cyclodextrinase | - |
Massilia timonae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
- |
Massilia timonae |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 50 | the enzyme remains stable after 30 min incubation at 45°C and loses all its activity when incubated for 30 min at 50°C. The temperature at which the enzyme loses 50% of its activity with 2 mM calcium is 48.5°C. The enzyme remains 60% active after 24 h at 25 or 40°C but is 20% active at 0°C | Massilia timonae |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 138.8 | - |
beta-cyclodextrin | at pH 7.5 and 40°C | Massilia timonae | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Massilia timonae |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 8 | the enzyme remains 100% active when incubated at pH values 6.5 and 8.0 after 24 h of incubation, and is 60% active when incubated at pH 7.0 for 24 h | Massilia timonae |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 68.3 | - |
beta-cyclodextrin | at pH 7.5 and 40°C | Massilia timonae | |
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