EC Number | Cloned (Comment) | Organism |
---|---|---|
4.1.1.7 | wild-type and mutant enzymes A460I, F464I and A460I/F464I | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
4.1.1.1 | I472A/I476F | increase in substrate binding affinity and specificity, highest enantioselectivity for (S)-acetoin, very low yield of product | Zymomonas mobilis |
4.1.1.1 | I476F | rapid loss of cofactor thiamine diphosphate. Improvement of enantioselectivity for (S)-acetoin | Zymomonas mobilis |
4.1.1.1 | mutant I472A | 2fold decrease in pyruvate decarboxylase activity, switch in substrate specificity to catalyse decarboxylation of benzoylformate, chimera between pyruvate decarboxylase and benzoylformate decarboxylase. Preferred substrates are 2-ketopentanoic acid and 2-ketohexanoic acid. Improvement of enantioselectivity for (S)-acetoin | Zymomonas mobilis |
4.1.1.7 | A460I | activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 2.3fold higher than wild-type activity, activity with 2-oxopentanoic acid is 1.1fold lower than wild-type activity, activity with 2-oxohexanoic acid is 3.3fold higher than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.3fold higher than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.1fold lower than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.3fold higher than wild-type activity, activity with 4,4-dimethyl-2-oxopentanoic acid is identical to wild-type activity, activity with 2-oxo-4-methylhexanoic acid is 2.2fold higher than wild-type activity, activity with benzoylformate is 22.5fold lower than wild-type activity, activity with 2-oxo-3-phenylpropanoic acid is identical to wild-type activity, activity with 2-oxo-4-phenylbutanoic acid is 1.8fold higher than wild-type activity, activity with 2-oxo-5-phenylpentanoic acid is identical to wild-type activity. The ratio of turnover number to KM-value for 2-oxo-4-methylpentanoic acid is 10fold higher than the wild-type value, the ratio of turnover number to KM-value for 2-oxo-4-methylhexanoic acid is 1.13fold higher than the wild-type value, the ratio of turnover number to KM-value for benzoylformate is 5.4fold lower than the wild-type value, the ratio of turnover number to KM-value for 2-oxohexanoic acid is 14.1fold higher than the wild-type value, the ratio of turnover number to KM-value for 2-oxopentanoic acid is 4.6fold higher than the wild-type value. Mutation has no effect on the range of products obtained by carboligation of acetaldehyde and benzaldehyde, the yield of the product 2-hydroxypropiophenone an decreases about 3fold and the enantioselectivity of acetoin and 2-hydroxypropiophenone is altered | Pseudomonas putida |
4.1.1.7 | A460I/F464I | activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 1.8fold lower than wild-type activity, activity with 2-oxopentanoic acid is 9.2fold lower than wild-type value, activity with 2-oxohexanoic acid is 3.3fold lower than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.5fold lower than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.2fold lower than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.5fold lower than wild-type activity, activity with 4,4-dimethyl-2-oxopentanoic acid is 10fold lower than wild-type activity, activity with 2-oxo-4-methylhexanoic acid is 1.1fold lower than wild-type activity, activity with benzoylformate is 64.3fold lower than wild-type activity, no activity with 2-oxo-3-phenylpropanoic acid, activity with 2-oxo-4-phenylbutanoic acid is 1.3fold lower than wild-type activity, activity with 2-oxo-5-phenylpentanoic acid is fold higher than wild-type activity. the ratio of turnover number to KM-value for benzoylformate is 88.7fold lower than the wild-type value | Pseudomonas putida |
4.1.1.7 | G464I | activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 1.2fold lower than wild-type activity, activity with 2-oxopentanoic acid is 4.8fold lower than wild-type value, activity with 2-oxohexanoic acid is 1.6fold lower than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.1fold higher than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.3fold higher than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.1fold higher than wild-type activity, activity with 4,4-dimethyl-2-oxopentanoic acid is identical to wild-type activity, activity with 2-oxo-4-methylhexanoic acid is 1.3fold higher than wild-type activity, activity with, activity with benzoylformate is 9.2fold lower than wild-type activity, activity with 2-oxo-3-phenylpropanoic acid is identical to wild-type activity, activity with 2-oxo-4-phenylbutanoic acid is9.3fold lower than wild-type activity, no activity with 2-oxo-5-phenylpentanoic acid. The ratio of turnover number to KM-value for benzoylformate is 31.8fold lower than the wild-type value. Mutation has no effect on the range of products obtained by carboligation of acetaldehyde and benzaldehyde, the yield of the product 2-hydroxypropiophenone an decreases about 3fold and the enantioselectivity of acetoin and 2-hydroxypropiophenone is altered | Pseudomonas putida |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
4.1.1.7 | 2-oxo-4-methylhexanoic acid | substrate inhibition | Pseudomonas putida | |
4.1.1.7 | 2-oxohexanoic acid | substrate inhibition | Pseudomonas putida | |
4.1.1.7 | 2-oxopentanoic acid | substrate inhibition | Pseudomonas putida |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.1.1.1 | 0.2 | - |
2-ketohexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 0.5 | - |
2-ketohexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 1.1 | - |
pyruvate | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 1.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 1.8 | - |
benzoylformate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 2.5 | - |
2-ketopentanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 2.6 | - |
pyruvate | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 3.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 4.4 | - |
benzoylformate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 4.7 | - |
2-ketobutanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 6.7 | - |
2-ketobutanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 7.6 | - |
2-ketopentanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 7.8 | - |
pyruvate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 11 | - |
2-ketopentanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 12.7 | - |
2-ketohexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 40 | - |
2-keto-4-methylhexanoic acid | and above, wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 50 | - |
pyruvate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 50 | - |
2-ketobutanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.7 | additional information | - |
additional information | KM-value of wild-type enzyme for 2-oxo-4-methylhexanoic acid (pH 6.0, 30°C) is above 20 mM | Pseudomonas putida | |
4.1.1.7 | 0.2 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 0.8 | - |
benzoylformate | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 1 | - |
2-oxohexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 1.1 | - |
2-oxo-4-methylhexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 1.1 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme A460I/F464I | Pseudomonas putida | |
4.1.1.7 | 1.2 | - |
2-oxopentanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 2.3 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme F464I | Pseudomonas putida | |
4.1.1.7 | 3.4 | - |
2-oxo-4-methylpentanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 4.1 | - |
2-oxohexanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 6 | - |
2-oxopentanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 9.6 | - |
2-oxo-4-methylpentanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 18.3 | - |
2-oxobutanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
4.1.1.1 | Mg2+ | required | Zymomonas mobilis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.1.1.1 | Zymomonas mobilis | P06672 | - |
- |
4.1.1.7 | Pseudomonas putida | P20906 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.1.1.7 | recombinant wild-type and mutant enzymes | Pseudomonas putida |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.1.1.1 | 2-keto-4-methylhexanoic acid | - |
Zymomonas mobilis | 3-methylpentanal + CO2 | - |
? | |
4.1.1.1 | 2-ketobutanoic acid | - |
Zymomonas mobilis | propanal + CO2 | - |
? | |
4.1.1.1 | 2-ketohexanoic acid | - |
Zymomonas mobilis | pentanal + CO2 | - |
? | |
4.1.1.1 | 2-ketopentanoic acid | - |
Zymomonas mobilis | butanal + CO2 | - |
? | |
4.1.1.1 | benzoylformate | substrate only for mutant mutant I472A | Zymomonas mobilis | benzaldehyde + CO2 | - |
? | |
4.1.1.1 | pyruvate | - |
Zymomonas mobilis | acetaldehyde + CO2 | - |
? | |
4.1.1.7 | 2-oxo-3-phenylpropanoic acid | - |
Pseudomonas putida | phenylacetaldehyde + CO2 | - |
? | |
4.1.1.7 | 2-oxo-4-methylhexanoic acid | - |
Pseudomonas putida | 3-methylpentanal + CO2 | - |
? | |
4.1.1.7 | 2-oxo-4-methylpentanoic acid | - |
Pseudomonas putida | 3-methylbutanal + CO2 | - |
? | |
4.1.1.7 | 2-oxo-4-phenylbutanoic acid | - |
Pseudomonas putida | 3-phenylpropionaldehyde + CO2 | - |
? | |
4.1.1.7 | 2-oxo-5-phenylpentanoic acid | - |
Pseudomonas putida | 4-phenylpentanal + CO2 | - |
? | |
4.1.1.7 | 2-oxobutanoic acid | - |
Pseudomonas putida | propionaldehyde + CO2 | - |
? | |
4.1.1.7 | 2-oxohexanoic acid | - |
Pseudomonas putida | valeraldehyde + CO2 | - |
? | |
4.1.1.7 | 2-oxopentanoic acid | - |
Pseudomonas putida | butyraldehyde + CO2 | - |
? | |
4.1.1.7 | 3 benzaldehyde + acetaldehyde | - |
Pseudomonas putida | (R)-benzoin + (S)-2-hydroxypropiophenone | - |
? | |
4.1.1.7 | 3-methyl-2-oxobutanoic acid | - |
Pseudomonas putida | 2-methylpropanal + CO2 | - |
? | |
4.1.1.7 | 3-methyl-2-oxopentanoic acid | - |
Pseudomonas putida | 2-methylbutanal + CO2 | - |
? | |
4.1.1.7 | 4,4-dimethyl-2-oxopentanoic acid | - |
Pseudomonas putida | 3,3-dimethylbutanal + CO2 | - |
? | |
4.1.1.7 | benzoylformate | best substrate for wild-type enzyme and mutant enzymes A460I, F464I, and A4670I, F464I | Pseudomonas putida | benzaldehyde + CO2 | - |
? | |
4.1.1.7 | additional information | no activity with 3,3-dimethyl-2-oxopentanoic acid and cyclohexaneglyoxylic acid (wild-type enzyme and mutant enzymes A460I, F464I and A460I/F464I) | Pseudomonas putida | ? | - |
? | |
4.1.1.7 | pyruvate | activity of wild-type enzyme is very low, very low activity with mutant enzymes A460I, F464I, and A4670I, F464I | Pseudomonas putida | acetaldehyde + CO2 | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.1.1.7 | tetramer | - |
Pseudomonas putida |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.1.1.1 | 0.4 | - |
2-keto-4-methylhexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 0.8 | - |
2-ketohexanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 1.2 | - |
benzoylformate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 4 | - |
2-ketohexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 6.9 | - |
benzoylformate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 7.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 8 | - |
pyruvate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 8.1 | - |
2-keto-4-methylhexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 9 | - |
2-ketobutanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 11 | - |
2-ketopentanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 30 | - |
2-ketopentanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 32 | - |
2-ketobutanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 53 | - |
2-ketopentanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 77 | - |
pyruvate | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 82 | - |
2-ketohexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 130 | - |
2-ketohexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 200 | - |
pyruvate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 220 | - |
2-ketopentanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 250 | - |
2-ketobutanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 320 | - |
2-ketobutanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.1 | 486 | - |
pyruvate | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
4.1.1.7 | 8.6 | - |
2-oxo-4-methylhexanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 9.8 | - |
2-oxo-4-methylhexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 11 | - |
2-oxo-4-methylpentanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 17.3 | - |
2-oxobutanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 18.8 | - |
2-oxobutanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 21 | - |
2-oxohexanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 21 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme A460I/F464I | Pseudomonas putida | |
4.1.1.7 | 38 | - |
2-oxo-4-methylpentanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 40.6 | - |
2-oxopentanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 44.3 | - |
2-oxopentanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 62 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 71.8 | - |
2-oxohexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 123 | - |
benzoylformate | pH 6.0, 30°C, mutant enzyme F464I | Pseudomonas putida | |
4.1.1.7 | 1349 | - |
benzoylformate | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
4.1.1.1 | thiamine diphosphate | - |
Zymomonas mobilis | |
4.1.1.7 | thiamine diphosphate | dependent on | Pseudomonas putida |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.1.1.7 | 2 | 5 | 2-oxohexanoic acid | pH 6.0, 30°C, wild-type enzyme | Pseudomonas putida | |
4.1.1.7 | 10 | - |
2-oxohexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 20 | - |
2-oxopentanoic acid | pH 6.0, 30°C, wild-type enzyme and mutant enzyme A460I | Pseudomonas putida | |
4.1.1.7 | 48 | - |
2-oxo-4-methylhexanoic acid | pH 6.0, 30°C, mutant enzyme A460I | Pseudomonas putida |