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Literature summary for 1.1.5.12 extracted from

  • Jiang, T.; Guo, X.; Yan, J.; Zhang, Y.; Wang, Y.; Zhang, M.; Sheng, B.; Ma, C.; Xu, P.; Gao, C.
    A bacterial multidomain NAD-independent D-lactate dehydrogenase utilizes flavin adenine dinucleotide and Fe-S clusters as cofactors and quinone as an electron acceptor for D-lactate oxidization (2017), J. Bacteriol., 199, e00342 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.5.12

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pseudomonas putida KT2440

Inhibitors

Inhibitors Comment Organism Structure
Cd2+ complete inhibition Pseudomonas putida KT2440
Co2+ complete inhibition Pseudomonas putida KT2440
Cu2+ complete inhibition Pseudomonas putida KT2440
EDTA partial inhibition Pseudomonas putida KT2440
Fe2+ complete inhibition Pseudomonas putida KT2440
Mn2+ complete inhibition Pseudomonas putida KT2440
Zn2+ complete inhibition Pseudomonas putida KT2440

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0041
-
 coenzyme Q10 cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
0.0095
-
 cytochrome c cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
0.031
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
0.048
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
0.0515
-
 cytochrome c cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
0.057
-
 (R)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
0.094
-
 (R)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
0.1
-
 D-2-hydroxybutanoate pH 7.4. 30°C Pseudomonas putida KT2440
0.17
-
 D-2-hydroxybutanoate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
1.04
-
 (S)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
1.38
-
 D-glycerate pH 7.4. 30°C Pseudomonas putida KT2440
1.67
-
 D-glycerate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
1.78
-
 (S)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
 Pseudomonas putida KT2440 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
K+ slight activation Pseudomonas putida KT2440

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
118000
-
 gel filtration Pseudomonas putida KT2440

Organism

Organism UniProt Comment Textmining
Pseudomonas putida KT2440 Q88DT2
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
44.5
-
 substrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida KT2440 pyruvate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
(R)-lactate + coenzyme Q10
-
 Pseudomonas putida KT2440 pyruvate + reduced coenzyme Q10
-
 ?
(R)-lactate + cytochrome c
-
 Pseudomonas putida KT2440 pyruvate + reduced cytochrome c
-
 ?
(S)-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide poor substrate Pseudomonas putida KT2440 pyruvate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
D-2-hydroxybutanoate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida KT2440 2-oxobutanoate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
D-glycerate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide poor substrate Pseudomonas putida KT2440 D-glyceraldehyde + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
additional information quinone is the preferred electron acceptor of the enzyme. No substrates: glycolate, L-2-hydroxybutyrate, D-mandelate, L-mandelate, D-3-phenyllactate, L-glycerate, DL-2-hydroxyisocaproate, and DL-2-hydroxyoctanoate Pseudomonas putida KT2440 ?
-
-

Subunits

Subunits Comment Organism
monomer 1 * 100000, SDS-PAGE Pseudomonas putida KT2440

Synonyms

Synonyms Comment Organism
D-iLDH
-
 Pseudomonas putida KT2440
Dld2
-
 Pseudomonas putida KT2440
pp4737
-
 Pseudomonas putida KT2440

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.49
-
 cytochrome c cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
0.78
-
 D-glycerate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
1.5
-
 (S)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
2
-
 cytochrome c cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
2.4
-
 coenzyme Q10 cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
5.3
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
8
-
 D-glycerate pH 7.4. 30°C Pseudomonas putida KT2440
9.5
-
 (S)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
13
-
 D-2-hydroxybutanoate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
17
-
 (R)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
66
-
 D-2-hydroxybutanoate pH 7.4. 30°C Pseudomonas putida KT2440
120
-
 (R)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
128
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440

Cofactor

Cofactor Comment Organism Structure
FAD the ratio of Fe-S D-iLDH to FAD is 0.92. Compared to the intact enzyme, the FAD-containing dehydrogenase domain shows increased catalytic efficiency with cytochrome c as the electron acceptor, but it completely lost the ability to use coenzyme Q10. The isolated FAD-containing dehydrogenase domain is no longer associated with the cell membrane, and it can not support the utilization of D-lactate as a carbon source Pseudomonas putida KT2440
Fe-S center protein is predicted to bind three [4Fe-4S] clusters and a [3Fe-4S] cluster overall Pseudomonas putida KT2440

General Information

General Information Comment Organism
metabolism the isolated FAD-containing dehydrogenase domain retains 2-hydroxyacid-oxidizing activity, although it decreased compared to the full Fe-S D-iLDH. Compared to the intact enzyme, the FAD-containing dehydrogenase domain shows increased catalytic efficiency with cytochrome c as the electron acceptor, but it completely lost the ability to use coenzyme Q10. The Fe-S oxidoreductase domain functions as an electron transfer component to facilitate the utilization of quinone as an electron acceptor by Fe-S D-iLDH, and it helps the enzyme associate with the cell membrane Pseudomonas putida KT2440

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.47
-
 D-glycerate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
0.84
-
 (S)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
6
-
 D-glycerate pH 7.4. 30°C Pseudomonas putida KT2440
9
-
 (S)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
9.5
-
 cytochrome c cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
78
-
 D-2-hydroxybutanoate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
170
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
176
-
 (R)-lactate isolated FAD-containing dehydrogenase domain, pH 7.4. 30°C Pseudomonas putida KT2440
210
-
 cytochrome c cosubstrate (R)-lactate, isolated FAD-containing dehydrogenase domain, pH 7.4, 30°C Pseudomonas putida KT2440
590
-
 coenzyme Q10 cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440
660
-
 D-2-hydroxybutanoate pH 7.4. 30°C Pseudomonas putida KT2440
2100
-
 (R)-lactate pH 7.4. 30°C Pseudomonas putida KT2440
2700
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cosubstrate (R)-lactate, pH 7.4, 30°C Pseudomonas putida KT2440