Activating Compound | Comment | Organism | Structure |
---|---|---|---|
dithiothreitol | 154% activity at 20 mM | Lactococcus lactis | |
dithiothreitol | 20 mM, 1.5fold activation | Lactococcus cremoris | |
EDTA | 160% activity at 10 mM | Lactococcus lactis | |
EDTA | 10 mM, 1.6fold activation | Lactococcus cremoris |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta cells | Lactococcus lactis |
overexpression in Escherichia coli as His-tagged fusion protein | Lactococcus cremoris |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 72% residual activity at 1 mM | Lactococcus lactis | |
CaCl2 | 1 mM, 28% inhibition | Lactococcus cremoris | |
Cu2+ | complete inhibition at 0.1 mM | Lactococcus lactis | |
CuSO4 | 0.1 mM, complete inhibition | Lactococcus cremoris | |
DEPC | 65% residual activity at 1 mM, 12% residual activity at 2 mM | Lactococcus lactis | |
diethyl dicarbonate | 2 mM, 88% inhibition | Lactococcus cremoris | |
Hg2+ | complete inhibition at 0.1 mM | Lactococcus lactis | |
HgCl2 | 0.1 mM, complete inhibition | Lactococcus cremoris | |
iodoacetamide | 0.1 mM, 5% inhibition | Lactococcus cremoris | |
iodoacetamide | 95% residual activity at 1 mM | Lactococcus lactis | |
Mg2+ | 83% residual activity at 1 mM | Lactococcus lactis | |
MgCl2 | 1 mM, 17% inhibition | Lactococcus cremoris |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
NADH | at pH 7.0 and 37°C | Lactococcus lactis | |
0.21 | - |
3-methyl-2-oxopentanoate | pH 7.0, 37°C | Lactococcus cremoris | |
0.21 | - |
2-oxomethylvalerate | at pH 7.0 and 37°C | Lactococcus lactis | |
0.29 | - |
3-methyl-2-oxobutanoate | pH 7.0, 37°C | Lactococcus cremoris | |
0.29 | - |
2-oxoisovalerate | at pH 7.0 and 37°C | Lactococcus lactis | |
0.3 | - |
4-methyl-2-oxopentanoate | pH 7.0, 37°C | Lactococcus cremoris | |
0.3 | - |
2-oxoisocaproate | at pH 7.0 and 37°C | Lactococcus lactis | |
0.53 | - |
2-oxovalerate | at pH 7.0 and 37°C | Lactococcus lactis | |
1.26 | - |
2-formylbutanethioate | pH 7.0, 37°C | Lactococcus cremoris | |
1.26 | - |
4-methylthio-2-oxobutanoate | at pH 7.0 and 37°C | Lactococcus lactis | |
1.5 | - |
benzoylformate | pH 7.0, 37°C | Lactococcus cremoris | |
1.5 | - |
benzoylformate | at pH 7.0 and 37°C | Lactococcus lactis | |
2.4 | - |
2-oxocaproate | at pH 7.0 and 37°C | Lactococcus lactis | |
3 | - |
D-mandelate | at pH 7.0 and 37°C | Lactococcus lactis | |
3 | - |
DL-2-hydroxyisocaproate | at pH 7.0 and 37°C | Lactococcus lactis | |
5.4 | - |
phenylpyruvate | pH 7.0, 37°C | Lactococcus cremoris | |
5.4 | - |
phenylpyruvate | at pH 7.0 and 37°C | Lactococcus lactis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34361 | - |
2 * 34361, calculated from amino acid sequence | Lactococcus lactis |
36000 | - |
2 * 36000, SDS-PAGE | Lactococcus cremoris |
36000 | - |
2 * 36000, His-tagged fusion protein, SDS-PAGE | Lactococcus lactis |
70000 | - |
gel filtration | Lactococcus lactis |
70000 | - |
gel filtration | Lactococcus cremoris |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Lactococcus cremoris | - |
- |
- |
Lactococcus cremoris TIL46 | - |
- |
- |
Lactococcus lactis | - |
- |
- |
Lactococcus lactis IL1403 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Lactococcus cremoris |
Storage Stability | Organism |
---|---|
4°C, concentrated enzyme solution at pH 7.0 (10.6 mg/ml), 3 months, without significant loss of activity | Lactococcus lactis |
4°C, diluted enzyme solution at pH 7.0 (0.004 mg/ml), 1 h, 40% loss of activity | Lactococcus lactis |
4°C, pH 7.0, concentrated enzyme solution (10.6 mg/ml) is stable for at least 2-3 months | Lactococcus cremoris |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-formylbutanethioate + NADH + H+ | i.e. 2-ketomethylthiobutyrate | Lactococcus cremoris | ? | - |
? | |
2-formylbutanethioate + NADH + H+ | i.e. 2-ketomethylthiobutyrate | Lactococcus cremoris TIL46 | ? | - |
? | |
2-oxobutyrate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxobutyrate + NADH + H+ | - |
Lactococcus lactis IL1403 | ? + NAD+ | - |
r | |
2-oxocaproate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxocaproate + NADH + H+ | - |
Lactococcus lactis IL1403 | ? + NAD+ | - |
r | |
2-oxoisocaproate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxoisocaproate + NADH + H+ | - |
Lactococcus lactis IL1403 | ? + NAD+ | - |
r | |
2-oxoisovalerate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxomethylthiobutyrate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxomethylvalerate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
2-oxovalerate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
3-methyl-2-oxobutanoate + NADH + H+ | i.e.2-oxoisovalerate | Lactococcus cremoris | 2-hydroxy-3-methylbutanoate + NAD+ | - |
? | |
3-methyl-2-oxobutanoate + NADH + H+ | i.e.2-oxoisovalerate | Lactococcus cremoris TIL46 | 2-hydroxy-3-methylbutanoate + NAD+ | - |
? | |
3-methyl-2-oxopentanoate + NADH + H+ | i.e. 2-oxomethylvalerate | Lactococcus cremoris | 2-hydroxy-3-methylpentanoate + NAD+ | - |
? | |
3-methyl-2-oxopentanoate + NADH + H+ | i.e. 2-oxomethylvalerate | Lactococcus cremoris TIL46 | 2-hydroxy-3-methylpentanoate + NAD+ | - |
? | |
4-methyl-2-oxopentanoate + NADH + H+ | i.e. 2-oxoisocaproate. The apparent Vmax/Km ratio for the reverse reaction is about 0.5% of that for the forward reaction | Lactococcus cremoris | (R)-2-hydroxy-4-methylpentanoate + NAD+ | i.e. (R)-2-hydroxyisocaproate | r | |
4-methyl-2-oxopentanoate + NADH + H+ | i.e. 2-oxoisocaproate. The apparent Vmax/Km ratio for the reverse reaction is about 0.5% of that for the forward reaction | Lactococcus cremoris TIL46 | (R)-2-hydroxy-4-methylpentanoate + NAD+ | i.e. (R)-2-hydroxyisocaproate | r | |
benzoylformate + NADH + H+ | - |
Lactococcus cremoris | ? | - |
? | |
benzoylformate + NADH + H+ | - |
Lactococcus cremoris TIL46 | ? | - |
? | |
benzoylformate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
D-mandelate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
DL-2-hydroxyisocaproate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
additional information | the enzyme accepts D-2-hydroxyacids but not L-2-hydroxyacids and shows no NADP-dependent 2-ketopantoate reductase activity. No reactions are observed with 10 mM L-2-hydroxyisocaproate and 1 mM NAD+, 10 mM pyruvate and 0.3 mM NADH, 10 mM 2-oxooisocaproate and 0.3 mM NADPH, and 10 mM 2-oxopantoate and 0.3 mM NADH or NADPH | Lactococcus lactis | ? | - |
? | |
additional information | the enzyme accepts D-2-hydroxyacids but not L-2-hydroxyacids and shows no NADP-dependent 2-ketopantoate reductase activity. No reactions are observed with 10 mM L-2-hydroxyisocaproate and 1 mM NAD+, 10 mM pyruvate and 0.3 mM NADH, 10 mM 2-oxooisocaproate and 0.3 mM NADPH, and 10 mM 2-oxopantoate and 0.3 mM NADH or NADPH | Lactococcus lactis IL1403 | ? | - |
? | |
phenylpyruvate + NADH + H+ | - |
Lactococcus lactis | ? + NAD+ | - |
r | |
phenylpyruvate + NADH + H+ | - |
Lactococcus lactis IL1403 | ? + NAD+ | - |
r | |
phenylpyruvate + NADH + H+ | - |
Lactococcus cremoris | phenyllactate + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 36000, SDS-PAGE | Lactococcus cremoris |
homodimer | 2 * 34361, calculated from amino acid sequence | Lactococcus lactis |
homodimer | 2 * 36000, His-tagged fusion protein, SDS-PAGE | Lactococcus lactis |
Synonyms | Comment | Organism |
---|---|---|
D-2-hydroxyacid dehydrogenase | - |
Lactococcus lactis |
D-HicDH | - |
Lactococcus lactis |
HdhD | - |
Lactococcus cremoris |
PanE | - |
Lactococcus lactis |
PanE | formerly | Lactococcus cremoris |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Lactococcus cremoris |
55 | - |
- |
Lactococcus lactis |
55 | - |
- |
Lactococcus cremoris |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 55 | at 37°C, the activity reaches about 65% of peak activity at 55°C | Lactococcus lactis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | 7 | forward reaction | Lactococcus lactis |
7 | - |
assay at | Lactococcus cremoris |
9 | - |
reverse reaction | Lactococcus lactis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | activity is reduced by only about 10% at pH 5.0 and 8.0 | Lactococcus lactis |
5.5 | 7 | reduction of 4-methyl-2-oxopentanoate | Lactococcus cremoris |
9 | - |
oxidation of (R)-2-hydroxy-4-methylpentanoate | Lactococcus cremoris |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | activity is reduced by about 10% at pH 5.0 and 8.0 | Lactococcus cremoris |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | NADH is the exclusive coenzyme | Lactococcus lactis | |
NADH | NADPH shows no activity as cosubstrate | Lactococcus cremoris |
General Information | Comment | Organism |
---|---|---|
malfunction | the inactivation of panE does not affect the total percentage of leucine degraded but totally prevents KIC reduction to 2-hydroxyisocaproate and slightly decreases the production of isovalerate | Lactococcus lactis |
malfunction | the inactivation of panE does not affect the total percentage of leucine degraded but totally prevented 4-methyl-2-oxopentanoate reduction to 2-hydroxyisocaproate and slightly decreased the production of isovalerate | Lactococcus cremoris |
metabolism | its probable physiological role is to regenerate the NAD+ necessary to catabolize branched-chain amino acids, leading to the production of ATP and aroma compounds responsible for the reduction of the 2-keto acids derived from leucine, isoleucine, and valine | Lactococcus cremoris |