Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli Rosetta (DE3) | Saccharolobus solfataricus |
Protein Variants | Comment | Organism |
---|---|---|
I336R/Y388H | enhanced enzyme activity | Saccharolobus solfataricus |
V203N/I336R | enhanced enzyme activity | Saccharolobus solfataricus |
V203N/I336R/Y388H | enhanced enzyme activity, 21fold higher enzyme efficiency (kcat/KM) compared to the wildtype enzyme. Biotransformation reaches more than 49% conversion and more than 99% enantiomeric excess at 80 °C after 2 h (with the same condition, the wild-type enzyme reached 12.3% conversion). Optimal temperature is 10°C lower than the optimum of the wild-type enzyme | Saccharolobus solfataricus |
V203N/Y388H | enhanced enzyme activity | Saccharolobus solfataricus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Co2+ | inhibition on the enzyme activity of both the wild-type and mutant proteins | Saccharolobus solfataricus | |
Cu2+ | inhibition on the enzyme activity of both the wild-type and mutant proteins | Saccharolobus solfataricus | |
Zn2+ | inhibition on the enzyme activity of both the wild-type and mutant proteins | Saccharolobus solfataricus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
17.5 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H/I336R | Saccharolobus solfataricus | |
22 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H | Saccharolobus solfataricus | |
26.5 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/I336R | Saccharolobus solfataricus | |
55.9 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme I336R | Saccharolobus solfataricus | |
60.8 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme Y388H/I336R | Saccharolobus solfataricus | |
61.3 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, wild-type enzyme | Saccharolobus solfataricus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | slightly enhances activity of mutant enzyme V203N/I336R | Saccharolobus solfataricus | |
Mn2+ | slightly enhances activity of mutant enzyme V203N/I336R | Saccharolobus solfataricus | |
Sn2+ | slightly enhances activity of mutant enzyme V203N/I336R | Saccharolobus solfataricus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | P95896 | - |
- |
Saccharolobus solfataricus ATCC 35092 | P95896 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharolobus solfataricus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one + H2O | - |
Saccharolobus solfataricus | + (+)-4-amino-cyclopent-2-enecarboxylic acid | - |
? | |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one + H2O | - |
Saccharolobus solfataricus ATCC 35092 | + (+)-4-amino-cyclopent-2-enecarboxylic acid | - |
? | |
2-azabicyclo[2.2.1]hept-5-en-3-one + H2O | - |
Saccharolobus solfataricus | (-)-2-azabicyclo[2.2.1]hept-5-en-3-one + (+)-4-amino-cyclopent-2-enecarboxylic acid | - |
? | |
2-azabicyclo[2.2.1]hept-5-en-3-one + H2O | - |
Saccharolobus solfataricus ATCC 35092 | (-)-2-azabicyclo[2.2.1]hept-5-en-3-one + (+)-4-amino-cyclopent-2-enecarboxylic acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Sslact | - |
Saccharolobus solfataricus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Saccharolobus solfataricus |
80 | - |
mutant enzyme V203N/Y388H/I336R | Saccharolobus solfataricus |
90 | - |
wild-type enzyme and mutant enzymes I336R, I336H, V203N/I336R and Y388H/I366R | Saccharolobus solfataricus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | 100 | 60°C: about 45% of maximal activity, 100°C: about 55% of maximal activity, mutant enzyme I336R | Saccharolobus solfataricus |
60 | 100 | 60°C: about 50% of maximal activity, 90°C: about 75% of maximal activity, 100°C: about 30% of maximal activity, mutant enzyme V203N/I336R/Y388H | Saccharolobus solfataricus |
80 | 100 | 80°C: about 80% of maximal activity, 100°C: about 75% of maximal activity | Saccharolobus solfataricus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.41 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, wild-type enzyme | Saccharolobus solfataricus | |
1.54 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H | Saccharolobus solfataricus | |
4.76 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme I336R | Saccharolobus solfataricus | |
7.69 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme Y388H/I336R | Saccharolobus solfataricus | |
8.06 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/I336R | Saccharolobus solfataricus | |
9.04 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H/I336R | Saccharolobus solfataricus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Saccharolobus solfataricus |
8 | - |
wild-type enzyme | Saccharolobus solfataricus |
9 | - |
mutant enzyme V203N/I336R/Y388H | Saccharolobus solfataricus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | pH 6.0: about 70% of maximal activity, pH 10.0: about 45% of maximal activity, wild-type enzyme | Saccharolobus solfataricus |
6 | 10 | pH 6.0: about 90% of maximal activity, pH 10.0: about 80% of maximal activity, mutant enzyme V203N/I336R/Y388H | Saccharolobus solfataricus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.023 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, wild-type enzyme | Saccharolobus solfataricus | |
0.07 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H | Saccharolobus solfataricus | |
0.085 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme I336R | Saccharolobus solfataricus | |
0.126 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme Y388H/I336R | Saccharolobus solfataricus | |
0.304 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/I336R | Saccharolobus solfataricus | |
0.517 | - |
(+)-2-azabicyclo[2.2.1]hept-5-en-3-one | 37°C, pH 7.8, mutant enzyme V203N/Y388H/I336R | Saccharolobus solfataricus |