Protein Variants | Comment | Organism |
---|---|---|
del I81/delK82 | phenylpyruvate is the only substrate which is converted at a significant catalytic rate by the mutant enzyme. In the publication this mutant is referred to as Ile100ADELTA/Lys100BDELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
del I81/delK82/delN87/delP88 | specific modifications in catalytic rates and substrate recognition | Weissella confusa |
del L 83 | deletion mutant shows an altered substrate specificity. In the publication this mutant is referred to as Leu101DELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
delI81 | drastic reductions in the catalytic activity for all tested substrates. In the publication this mutant is referred to as Ile100ADELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
delK82 | drastic reductions in the catalytic activity for all tested substrates. In the publication this mutant is referred to as Lys100BDELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
delN87 | deletion mutant shows an altered substrate specificity. In the publication this mutant is referred to as Asn105ADELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
delN87/delP88 | for the deletion mutant enzyme 2-oxo carboxylic acids branched at C4 are better substrates than 2-oxocaproate, the substrate with the best kcat,/KM ratio known for the wild-type enzyme.The mutation results in a 5.2fold increased catalytic efficiency towards 4-methyl-2-oxopentanoate compared to the wild-type enzyme. In the publication this mutant is referred to as Asn105ADELTA/Pro105BDELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
delP88 | deletion mutant shows an altered substrate specificity. In the publication this mutant is referred to as Pro105BDELTA, according to the numbering system of the dogfish L-lactate dehydrogenase coenzyme loop region | Weissella confusa |
additional information | the coenzyme loop, a functional element which is essential for catalysis and substrate specificity, is modified in order to identify the residues involved in the catalytic reaction and substrate specificity | Weissella confusa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.026 | - |
phenylpyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
0.067 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
0.071 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
0.1 | 1 | 2-oxocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
0.1 | 2 | phenylpyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
0.17 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
0.33 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
0.41 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
0.62 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
0.9 | 1 | 4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
1 | 4 | 4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
2 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
2 | 5 | pyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
2.8 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
2.8 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del L83 | Weissella confusa | |
2.8 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
3.3 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
3.5 | - |
pyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
5 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
5.8 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
6.4 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del L83 | Weissella confusa | |
9.3 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
11 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del L83 | Weissella confusa | |
18 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
18 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
19 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
19 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
27 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
30 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
43 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
50 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
62 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
110 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
140 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
200 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Weissella confusa | P14295 | i.e. Lactobacillus confusus | - |
Purification (Comment) | Organism |
---|---|
- |
Weissella confusa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxocaproate + NADH + H+ | - |
Weissella confusa | 2-hydroxycaproate + NAD+ | - |
r | |
4-methyl-2-oxopentanoate + NADH + H+ | - |
Weissella confusa | (S)-2-hydroxy-4-methylpentanoate + NAD+ | - |
r | |
phenylpyruvate + NADH + H+ | - |
Weissella confusa | 3-phenyllactate + NAD+ | - |
r | |
pyruvate + NADH + H+ | - |
Weissella confusa | L-lactate + NAD+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
L-2-hydroxyisocaproate dehydrogenase | - |
Weissella confusa |
L-HicDH | - |
Weissella confusa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Weissella confusa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
0.03 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
0.23 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
0.63 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
0.76 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
1.7 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
8 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
13 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
15 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
17 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
20 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
25 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
39 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
40 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
46 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
90 | - |
pyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
280 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
490 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
620 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
750 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
920 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
1000 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
1200 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
2400 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
2600 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
4200 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
4400 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
7300 | - |
phenylpyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
12000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
15000 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
19000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
21000 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
89000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
360000 | - |
2-oxocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
520000 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Weissella confusa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00013 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
0.00021 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
0.0038 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
0.025 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
0.035 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
0.092 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
0.32 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
0.6 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
0.63 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
0.68 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
0.91 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
0.92 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
1.4 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81 | Weissella confusa | |
1.8 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
8 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
8.4 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
15 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delK82 | Weissella confusa | |
26 | - |
pyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
160 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
380 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
790 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82 | Weissella confusa | |
2200 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
2900 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
4300 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del I81/delK82/delN87/delP88 | Weissella confusa | |
4300 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme del L 83 | Weissella confusa | |
6300 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
8700 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delP88 | Weissella confusa | |
16000 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
34000 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
36000 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
46000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87 | Weissella confusa | |
190000 | - |
4-methyl-2-oxopentanoate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
270000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme delN87/delP88 | Weissella confusa | |
280000 | - |
phenylpyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
3300000 | - |
2-oxocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa |