Protein Variants | Comment | Organism |
---|---|---|
F236S | phenylpyruvate displays the largest kcat of the tested substrates. All kcat values, with the exception of phenylpyruvate, are reduced, but the relative acceptance of phenylglyoxylate is greatly increased | Weissella confusa |
F236V | decrease in the second-order rate constants for all tested substrates. The kcat values for the smaller substrates decrease drastically. Phenylpyruvate is the favourite substrate (2-oxocaproate is the favourite substrate of the wild-type enzyme) | Weissella confusa |
G234V/G235D | the second-order rate constant decreased for most substrates with the exception of pyruvate, which reacts 2.5times faster in the mutant enzyme than in the wild-type enzyme. The catalysis of 2-oxoisocaproate is unaffected | Weissella confusa |
L239A | shift of enzyme specificity towards the substrates branched at C3 corresponding to an increase in the turnover numbers for 2-oxoisocaproate | Weissella confusa |
L239F | slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values | Weissella confusa |
L239M | KM values of all substrates of the enzyme variant are very similar to those of the wild-type enzyme, large improvement in the kcat values of the C4-branched substrates 2-oxoisocaproate and phenylpyruvate, decrease in the turnover number of 2-oxocaproate (the substrate favoured by the wild-type enzyme). Whereas in the wild-type enzyme the second-order rate constant for 2-oxoisocaproate is only 1% of that for the unbranched substrate, in the mutant enzyme the rate constant of 2-oxocaproate is only 18% of that for 2-oxoisocaproate | Weissella confusa |
L239M/T245A | the catalytic rates are reduced by several orders of magnitude and the KM values shows at least a 100fold increase for most substrates (with the exception of phenylglyoxylate). With respect to L239M the substrate specificity shifts towards keto-tert-leucine and with respect to T245A 2-oxoisocaproate and phenylpyruvate are more favoured | Weissella confusa |
L239W | slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values | Weissella confusa |
T245A | the catalytic rates are reduced by several orders of magnitude, relative shift of substrate specificity for keto-tert-leucine of more than 17 000 compared with the 2-oxocaproate (kcat/KM) | Weissella confusa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Weissella confusa | P14295 | i.e. Lactobacillus confusus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxocaproate + NADH + H+ | 2-oxocaproate is the best substrate for the wild-type enzyme | Weissella confusa | 2-hydroxycaproate + NAD+ | - |
r | |
2-oxoisocaproate + NADH + H+ | - |
Weissella confusa | 2-hydroxyisocaproate + NAD+ | - |
r | |
additional information | broad substrate specificity, utilizes a wide range of 2-oxo acids branched at the C4 atom | Weissella confusa | ? | - |
? | |
oxo-tert-leucine + NADH + H+ | - |
Weissella confusa | ? + NAD+ | - |
r | |
phenylglyoxylate + NADH + H+ | - |
Weissella confusa | ? + NAD+ | - |
r | |
phenylpyruvate + NADH + H+ | - |
Weissella confusa | 3-phenyllactate + NAD+ | - |
r | |
pyruvate + NADH + H+ | - |
Weissella confusa | L-lactate + NAD+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
L-HicDH | - |
Weissella confusa |
L-hydroxyisocaproate dehydrogenase | - |
Weissella confusa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
0.075 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
0.1 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
0.1 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
0.2 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
0.2 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
0.2 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
0.3 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
0.4 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
0.9 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
1.3 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
2 | 8 | 2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
6.58 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
9.5 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
20 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
20 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
22 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
25 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
27 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
32 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
51 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
52 | - |
oxo-tert-leucine | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
90 | - |
pyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
92 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
100 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
150 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
210 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
320 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
340 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
350 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
420 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
520 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
920 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
1000 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
2100 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
2400 | - |
2-oxoisocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
2400 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
2500 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
2500 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
3400 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
3500 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
4100 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
4400 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
5600 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
6400 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
6900 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
7300 | - |
phenylpyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
7600 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
7900 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
11000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
13000 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
15000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
18000 | - |
phenylglyoxylate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
360000 | - |
2-oxocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00526 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
0.01 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
0.012 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
0.0465 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
0.0469 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
0.0684 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
0.07 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
0.357 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
0.411 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
0.654 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
1.08 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
1.82 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
2.97 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
3.09 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
3.84 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
5.88 | - |
pyruvate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
6.06 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
6.13 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
10 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
13 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
15 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme L239M/T245A | Weissella confusa | |
16 | - |
oxo-tert-leucine | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
22.5 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
25.7 | - |
pyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
32.8 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
66.2 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
117 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme T245A | Weissella confusa | |
133 | - |
oxo-tert-leucine | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
158 | - |
phenylglyoxylate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
400 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
677 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
941 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
1460 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239F | Weissella confusa | |
1530 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
1880 | - |
phenylglyoxylate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
1920 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
2190 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
2420 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
2620 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
2670 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
2880 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
5060 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239W | Weissella confusa | |
7330 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239A | Weissella confusa | |
10000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme F236V | Weissella confusa | |
28700 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
35800 | - |
2-oxoisocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
37600 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
39100 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
49200 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme F236S | Weissella confusa | |
76000 | - |
2-oxocaproate | pH 7.0, 30°C, mutant enzyme G234V/G235D | Weissella confusa | |
203000 | - |
2-oxoisocaproate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
224000 | - |
phenylpyruvate | pH 7.0, 30°C, mutant enzyme L239M | Weissella confusa | |
281000 | - |
phenylpyruvate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa | |
3270000 | - |
2-oxocaproate | pH 7.0, 30°C, wild-type enzyme | Weissella confusa |