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Literature summary for 1.1.1.337 extracted from

  • Schütte, W.; Hummel, W.; Kula, M.R.
    L-2-Hydroxyisocaproate dehydrogenase - A new enzyme from Lactobacillus confusus for the stereospecific reduction of 2-ketocarboxylic acids (1984), Appl. Microbiol. Biotechnol., 19, 167-176.
No PubMed abstract available

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 10 mM, 15% inhibition Weissella confusa
4-chloromercuribenzoate weak inhibition Weissella confusa
iodoacetamide weak inhibition Weissella confusa
iodoacetate weak inhibition Weissella confusa
KCN weak inhibition Weissella confusa
additional information reduced glutathione (20 mM) has no effect. EDTA, 2,2'-bipyridyl in 1 mM and 10 mM final concentrations have no effect on enzymatic activity Weissella confusa
NADH inhibition above 0.24 mM Weissella confusa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
2-oxoisocaproate pH 7.0, 30°C Weissella confusa
0.065
-
2-oxoisovalerate pH 7.0, 30°C Weissella confusa
0.1
-
2-oxovalerate pH 7.0, 30°C Weissella confusa
0.1 2 phenylpyruvate pH 7.0, 30°C Weissella confusa
0.1 2 2-oxocaproate pH 7.0, 30°C Weissella confusa
0.17
-
2-oxooctanoate pH 7.0, 30°C Weissella confusa
0.45
-
2-oxobutyrate pH 7.0, 30°C Weissella confusa
0.6
-
2-Hydroxyisovalerate pH 8.0, 30°C Weissella confusa
0.62
-
2-hydroxyisocaproate pH 8.0, 30°C Weissella confusa
0.64
-
3-Phenyllactate pH 8.0, 30°C Weissella confusa
1.8
-
2-hydroxyvalerate pH 8.0, 30°C Weissella confusa
1.9
-
2-Hydroxyoctanoate pH 8.0, 30°C Weissella confusa
2.2
-
2-hydroxycaproate pH 8.0, 30°C Weissella confusa
4.5
-
pyruvate pH 7.0, 30°C Weissella confusa
100
-
L-lactate pH 8.0, 30°C Weissella confusa

Metals/Ions

Metals/Ions Comment Organism Structure
additional information at 1 mM no influence is observed with Mg2+, Ca2+, Co2+, Ni2+, and Cd2+ and very weak effects with Mn2+, Cu2+, and Zn2+ Weissella confusa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33000
-
4 * 33000, SDS-PAGE Weissella confusa
125000
-
gel filtration Weissella confusa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
a 2-oxocarboxylate + NADH + H+ Weissella confusa
-
(2S)-2-hydroxycarboxylate + NAD+
-
r

Organism

Organism UniProt Comment Textmining
Weissella confusa P14295 i.e. Lactobacillus confusus
-

Purification (Commentary)

Purification (Comment) Organism
-
Weissella confusa

Source Tissue

Source Tissue Comment Organism Textmining
culture condition maximum specific and volume activities are found at the end of the exponential growth phase, during further fermentation enzyme activity declines rapidly and is no longer detectable after 15 h Weissella confusa
-

Storage Stability

Storage Stability Organism
4°C, pH 6.5, concentrated enzyme solution, 10% loss of activity after 6 months Weissella confusa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxycaproate + NAD+
-
Weissella confusa 2-oxocaproate + NADH + H+
-
r
2-hydroxyisocaproate + NAD+ the initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima Weissella confusa 2-oxoisocaproate + NADH + H+
-
r
2-hydroxyisovalerate + NAD+
-
Weissella confusa 2-oxoisovalerate + NADH + H+
-
r
2-hydroxyoctanoate + NAD+
-
Weissella confusa 2-oxooctanoate + NADH + H+
-
?
2-hydroxyvalerate + NAD+
-
Weissella confusa 2-oxovalerate + NADH + H+
-
r
2-oxobutyrate + NADH + H+
-
Weissella confusa 2-hydroxybutyrate + NAD+
-
r
2-oxocaproate + NADH + H+
-
Weissella confusa 2-hydroxycaproate + NAD+
-
r
2-oxoisocaproate + NADH + H+ 2-oxoisocaproate is the best substrate with the lowest KM-valus of 0.065 mM. Tthe initial rate of the reduction of 2-oxoisocaproate is about seven times faster than the reverse reaction, the dehydrogenation of L-2-hydroxyisocaproate, measured in buffers at the respective pH optima Weissella confusa 2-hydroxyisocaproate + NAD+
-
r
2-oxoisovalerate + NADH + H+
-
Weissella confusa 2-hydroxyisovalerate + NAD+
-
r
2-oxooctanoate + NADH + H+
-
Weissella confusa 2-hydroxyoctanoate + NAD+
-
r
2-oxovalerate + NADH + H+
-
Weissella confusa 2-hydroxyvalerate + NAD+
-
r
3-phenyllactate + NAD+
-
Weissella confusa pyruvate + NADH + H+
-
r
a 2-oxocarboxylate + NADH + H+
-
Weissella confusa (2S)-2-hydroxycarboxylate + NAD+
-
r
a 2-oxocarboxylate + NADH + H+ various 2-oxocarboxylic acids are stereospecifically reduced to the corresponding (S)-2-hydroxycarboxylic acids. In the reverse reaction the NAD+-dependent dehydrogenation of L-2-hydroxycarboxylic acids is observed Weissella confusa (2S)-2-hydroxycarboxylate + NAD+
-
r
L-lactate + NAD+
-
Weissella confusa pyruvate + NADH + H+
-
r
phenylpyruvate + NADH + H+
-
Weissella confusa 3-phenyllactate + NAD+
-
r
pyruvate + NADH + H+
-
Weissella confusa L-lactate + NAD+
-
r

Subunits

Subunits Comment Organism
tetramer 4 * 33000, SDS-PAGE Weissella confusa

Synonyms

Synonyms Comment Organism
L-2-hydroxyisocaproate dehydrogenase
-
Weissella confusa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Weissella confusa
50
-
-
Weissella confusa

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 the maximal temperature is around 50°C. At this temperature the reaction proceeded at about 3fold increase of the rate compared to 30°C Weissella confusa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
1 h, stable up to Weissella confusa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
reduction of 2-oxoisocaproate Weissella confusa
8 8.5 dehydrogenation of L-2-hydroxyisocaproate Weissella confusa

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 9 4°C, 24 h, stable Weissella confusa
6.5
-
4°C, 6 months, concentrated enzyme solutions, 10% loss of activity Weissella confusa

Cofactor

Cofactor Comment Organism Structure
NAD+ the enzyme requires NAD+ as a cofactor, no reaction with NADP+ in concentrations up to 3 mM Weissella confusa
NADH the enzyme requires NADH as a cofactor, no reaction with NADPH up to 0.24 mM Weissella confusa