Application | Comment | Organism |
---|---|---|
synthesis | synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate in Escherichia coli. Coexpression of carbonyl reductase CRII and a glucose dehydrogenase gives an activity of 15 U/mg protein using ethyl 4-chloro-3-oxobutanoate as a substrate in a water/butyl acetate system. The transformants give a molar yield of 91%, and an optical purity of the (S)-isomer of more than 99% enantiomeric excess | Scheffersomyces stipitis |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Scheffersomyces stipitis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | 5 mM, 50% residual activity | Scheffersomyces stipitis | |
phenyl methyl sulfonylfluoride | 5 mM, 49% residual activity | Scheffersomyces stipitis | |
sodium dodecylsulfate | 1%, 51% residual activity | Scheffersomyces stipitis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.3 | - |
ethyl 4-chloro-3-oxobutanoate | pH 6.0, 30°C | Scheffersomyces stipitis |
Organic Solvent | Comment | Organism |
---|---|---|
dimethyl formamide | 1%, 56% residual activity | Scheffersomyces stipitis |
dimethyl sulfoxide | 1%, 67% residual activity | Scheffersomyces stipitis |
tetrahydrofuran | 1%, 77% residual activity | Scheffersomyces stipitis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Scheffersomyces stipitis | A3GF05 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Scheffersomyces stipitis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
224 | - |
pH 6.0, 30°C | Scheffersomyces stipitis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,3-butanedione + NADH + H+ | 9% of the activity with ethyl 4-chloro-3-oxobutanoate | Scheffersomyces stipitis | ? + NAD+ | - |
? | |
2,3-pentanedione + NADH + H+ | 82% of the activity with ethyl 4-chloro-3-oxobutanoate | Scheffersomyces stipitis | ? + NAD+ | - |
? | |
ethyl 2-chloro-3-oxobutanoate + NADH + H+ | 49% of the activity with ethyl 4-chloro-3-oxobutanoate | Scheffersomyces stipitis | ethyl 2-chloro-3-hydroxybutanoate + NAD+ | - |
? | |
ethyl 3-oxobutanoate + NADH + H+ | 28% of the activity with ethyl 4-chloro-3-oxobutanoate | Scheffersomyces stipitis | ethyl 3-hydroxybutanoate + NAD+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADH + H+ | - |
Scheffersomyces stipitis | ethyl (S)-4-chloro-3-hydroxybutanoate + NAD+ | product in more than 99% enantiomeric excess | ? |
Synonyms | Comment | Organism |
---|---|---|
CRII | - |
Scheffersomyces stipitis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Scheffersomyces stipitis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 35 | more than 80% of maximum activity within | Scheffersomyces stipitis |
50 | - |
no residual activity | Scheffersomyces stipitis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
- |
Scheffersomyces stipitis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 7 | more than 80% of maximum activity within | Scheffersomyces stipitis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5.6 | - |
between pH 5.6-7.0, 4°C, stable for 48 h | Scheffersomyces stipitis |
7 | - |
between pH 5.6-7.0, 4°C, stable for 48 h | Scheffersomyces stipitis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | preferred over NADPH | Scheffersomyces stipitis | |
NADPH | about 10% of the activity with NADH | Scheffersomyces stipitis |