Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutants in Escherichia coli BL21(DE3) | Alicyclobacillus acidocaldarius |
Protein Variants | Comment | Organism |
---|---|---|
M112S | site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms | Alicyclobacillus acidocaldarius |
M112S/R215L | site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms | Alicyclobacillus acidocaldarius |
M112T | site-directed and saturation mutagenesis, altered substrate specificity | Alicyclobacillus acidocaldarius |
M112V | site-directed and saturation mutagenesis, altered substrate specificity | Alicyclobacillus acidocaldarius |
R215L | site-directed and saturation mutagenesis, altered substrate specificity, higher efficiency with monoacylesters whose chain length is greater than 8 C-atoms | Alicyclobacillus acidocaldarius |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
HEPES | inhibition mechanism, competitive | Alicyclobacillus acidocaldarius | |
propan-2-ol | competitive | Alicyclobacillus acidocaldarius |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
4-nitrophenyl dodecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.008 | - |
4-nitrophenyl dodecanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.008 | - |
4-nitrophenyl decanoate | mutants M211S and M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.01 | - |
4-nitrophenyl dodecanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.011 | - |
4-nitrophenyl decanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.015 | - |
4-nitrophenyl dodecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.017 | - |
4-nitrophenyl tetradecanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.018 | - |
4-nitrophenyl decanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.019 | - |
4-nitrophenyl decanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.019 | - |
4-nitrophenyl tetradecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.019 | - |
4-nitrophenyl dodecanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.02 | - |
4-nitrophenyl tetradecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.021 | - |
4-nitrophenyl hexadecanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.021 | - |
4-nitrophenyl decanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.021 | - |
4-nitrophenyl dodecanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.022 | - |
4-nitrophenyl octanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.022 | - |
4-nitrophenyl tetradecanoate | mutants R215L and M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.023 | - |
4-nitrophenyl hexadecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.025 | - |
4-nitrophenyl octanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.026 | - |
4-nitrophenyl hexadecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.028 | - |
4-nitrophenyl octanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.031 | - |
4-nitrophenyl hexadecanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.031 | - |
4-nitrophenyl tetradecanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.034 | - |
4-nitrophenyl hexadecanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.037 | - |
4-nitrophenyl octanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.042 | - |
4-nitrophenyl hexadecanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.06 | - |
4-nitrophenyl octanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.09 | - |
4-nitrophenyl hexanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.099 | - |
4-nitrophenyl hexanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.1 | - |
4-nitrophenyl octanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.21 | - |
4-nitrophenyl hexanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.22 | - |
4-nitrophenyl hexanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.41 | - |
4-nitrophenyl hexanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.51 | - |
4-nitrophenyl hexanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.73 | - |
4-nitrophenyl butanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
0.75 | - |
4-nitrophenyl butanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1.06 | - |
4-nitrophenyl butanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1.27 | - |
4-nitrophenyl butanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1.77 | - |
4-nitrophenyl butanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1.86 | - |
4-nitrophenyl butanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alicyclobacillus acidocaldarius | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutants from Escherichia coli BL21(DE3) | Alicyclobacillus acidocaldarius |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a carboxylic ester + H2O = an alcohol + a carboxylate | reaction mechanism | Alicyclobacillus acidocaldarius |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + acetate | - |
r | |
4-nitrophenyl butanoate + H2O | best substrate of mutants M211V, M211T, M211S and M211S/R215L | Alicyclobacillus acidocaldarius | 4-nitrophenol + butanoate | - |
r | |
4-nitrophenyl decanoate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + decanoate | - |
r | |
4-nitrophenyl dodecanoate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + dodecanoate | - |
r | |
4-nitrophenyl hexadecanoate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + hexadecanoate | - |
r | |
4-nitrophenyl hexanoate + H2O | best substrate of the wild-type enzyme | Alicyclobacillus acidocaldarius | 4-nitrophenol + hexanoate | - |
r | |
4-nitrophenyl octanoate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + octanoate | - |
r | |
4-nitrophenyl tetradecanoate + H2O | - |
Alicyclobacillus acidocaldarius | 4-nitrophenol + tetradecanoate | - |
r | |
additional information | substrate specificities of wild-type and mutant enzymes, reverse reaction in performed in organic solvents | Alicyclobacillus acidocaldarius | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EST2 | - |
Alicyclobacillus acidocaldarius |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
- |
Alicyclobacillus acidocaldarius |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7 | - |
4-nitrophenyl hexadecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
8.3 | - |
4-nitrophenyl hexadecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
15.6 | - |
4-nitrophenyl hexadecanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
21 | - |
4-nitrophenyl hexadecanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
27 | - |
4-nitrophenyl tetradecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
35 | - |
4-nitrophenyl hexadecanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
40 | - |
4-nitrophenyl tetradecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
44 | - |
4-nitrophenyl hexadecanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
105 | - |
4-nitrophenyl tetradecanoate | wild-type enzyme and mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
110 | - |
4-nitrophenyl dodecanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
112 | - |
4-nitrophenyl dodecanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
232 | - |
4-nitrophenyl tetradecanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
235 | - |
4-nitrophenyl tetradecanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
350 | - |
4-nitrophenyl dodecanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
390 | - |
4-nitrophenyl dodecanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
465 | - |
4-nitrophenyl decanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
626 | - |
4-nitrophenyl decanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
705 | - |
4-nitrophenyl dodecanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
860 | - |
4-nitrophenyl decanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
940 | - |
4-nitrophenyl dodecanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
950 | - |
4-nitrophenyl decanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
970 | - |
4-nitrophenyl decanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1376 | - |
4-nitrophenyl octanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1380 | - |
4-nitrophenyl octanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1500 | - |
4-nitrophenyl hexanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1645 | - |
4-nitrophenyl octanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1650 | - |
4-nitrophenyl octanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1670 | - |
4-nitrophenyl decanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1740 | - |
4-nitrophenyl octanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1855 | - |
4-nitrophenyl octanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1860 | - |
4-nitrophenyl octanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
1960 | - |
4-nitrophenyl octanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
2180 | - |
4-nitrophenyl butanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
2300 | - |
4-nitrophenyl hexanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
2750 | - |
4-nitrophenyl butanoate | mutant M211S/R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
3080 | - |
4-nitrophenyl octanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
3200 | - |
4-nitrophenyl butanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
3420 | - |
4-nitrophenyl hexanoate | wild-type enzyme, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
3660 | - |
4-nitrophenyl hexanoate | mutant R215L, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
3890 | - |
4-nitrophenyl hexanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
4000 | - |
4-nitrophenyl butanoate | mutant M211V, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
4230 | - |
4-nitrophenyl butanoate | mutant M211S, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
4440 | - |
4-nitrophenyl hexanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
5240 | - |
4-nitrophenyl butanoate | mutant M211T, pH 7.1, 70°C | Alicyclobacillus acidocaldarius |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.1 | - |
- |
Alicyclobacillus acidocaldarius |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
130 | - |
propan-2-ol | competitive, pH 7.1, 70°C | Alicyclobacillus acidocaldarius | |
780 | - |
HEPES | competitive, pH 7.1, 70°C | Alicyclobacillus acidocaldarius |