Protein Variants | Comment | Organism |
---|---|---|
I472A/I476F | increase in substrate binding affinity and specificity, highest enantioselectivity for (S)-acetoin, very low yield of product | Zymomonas mobilis |
I476F | rapid loss of cofactor thiamine diphosphate. Improvement of enantioselectivity for (S)-acetoin | Zymomonas mobilis |
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 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
2-ketohexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
0.5 | - |
2-ketohexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
1.1 | - |
pyruvate | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
1.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
1.8 | - |
benzoylformate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
2.5 | - |
2-ketopentanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
2.6 | - |
pyruvate | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
3.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.4 | - |
benzoylformate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4.7 | - |
2-ketobutanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
6.7 | - |
2-ketobutanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
7.6 | - |
2-ketopentanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
7.8 | - |
pyruvate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
11 | - |
2-ketopentanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
12.7 | - |
2-ketohexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
40 | - |
2-keto-4-methylhexanoic acid | and above, wild type, pH 6.5, 30°C | Zymomonas mobilis | |
50 | - |
pyruvate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
50 | - |
2-ketobutanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Zymomonas mobilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Zymomonas mobilis | P06672 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-keto-4-methylhexanoic acid | - |
Zymomonas mobilis | 3-methylpentanal + CO2 | - |
? | |
2-ketobutanoic acid | - |
Zymomonas mobilis | propanal + CO2 | - |
? | |
2-ketohexanoic acid | - |
Zymomonas mobilis | pentanal + CO2 | - |
? | |
2-ketopentanoic acid | - |
Zymomonas mobilis | butanal + CO2 | - |
? | |
benzoylformate | substrate only for mutant mutant I472A | Zymomonas mobilis | benzaldehyde + CO2 | - |
? | |
pyruvate | - |
Zymomonas mobilis | acetaldehyde + CO2 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.4 | - |
2-keto-4-methylhexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
0.8 | - |
2-ketohexanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
1.2 | - |
benzoylformate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
4 | - |
2-ketohexanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
6.9 | - |
benzoylformate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
7.7 | - |
2-keto-4-methylhexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
8 | - |
pyruvate | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
8.1 | - |
2-keto-4-methylhexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
9 | - |
2-ketobutanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
11 | - |
2-ketopentanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
30 | - |
2-ketopentanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
32 | - |
2-ketobutanoic acid | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
53 | - |
2-ketopentanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
77 | - |
pyruvate | mutant I476F, pH 6.5, 30°C | Zymomonas mobilis | |
82 | - |
2-ketohexanoic acid | mutant I472A/I476F, pH 6.5, 30°C | Zymomonas mobilis | |
130 | - |
2-ketohexanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
200 | - |
pyruvate | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
220 | - |
2-ketopentanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
250 | - |
2-ketobutanoic acid | mutant I472A, pH 6.5, 30°C | Zymomonas mobilis | |
320 | - |
2-ketobutanoic acid | wild type, pH 6.5, 30°C | Zymomonas mobilis | |
486 | - |
pyruvate | wild type, pH 6.5, 30°C | Zymomonas mobilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
thiamine diphosphate | - |
Zymomonas mobilis |