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Literature summary for 1.1.5.10 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, e00342-17 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
heterologously expressed in Escherichia coli C43(DE3) as a His-tagged protein Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
antimycin A inhibits electron transfer from quinone to cytochrome c Pseudomonas putida
Ba2+ 5 mM, partial inhibition of activity Pseudomonas putida
Ca2+ 5 mM, partial inhibition of activity Pseudomonas putida
Cd2+ 5 mM, complete inhibition of activity Pseudomonas putida
Co2+ 5 mM, complete inhibition of activity Pseudomonas putida
Cu2+ 5 mM, complete inhibition of activity Pseudomonas putida
EDTA 5 mM, partial inhibition of activity Pseudomonas putida
Fe2+ 5 mM, complete inhibition of activity Pseudomonas putida
Mg2+ 5 mM, partial inhibition of activity Pseudomonas putida
Mn2+ 5 mM, complete inhibition of activity Pseudomonas putida
NaN3 inhibits electron transfer from cytochrome c to molecular oxygen Pseudomonas putida
Ni2+ 5 mM, partial inhibition of activity Pseudomonas putida
Zn2+ 5 mM, complete inhibition of activity Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0041
-
Ubiquinone-10 pH 7.4, 30°C Pseudomonas putida
0.048
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide pH 7.4, 30°C Pseudomonas putida
0.057
-
D-lactate pH 7.4, 30°C Pseudomonas putida
0.1
-
D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
1.04
-
L-lactate pH 7.4, 30°C Pseudomonas putida
1.38
-
D-glycerate pH 7.4, 30°C Pseudomonas putida

Localization

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

Metals/Ions

Metals/Ions Comment Organism Structure
K+ 5 mM, slightly positive effect Pseudomonas putida

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Pseudomonas putida Q88DT2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-2-hydroxybutyrate + ubiquinone-10
-
Pseudomonas putida 2-oxobutyrate + ubiquinol-10
-
?
D-glycerate + ubiquinone-10
-
Pseudomonas putida ? + ubiquinol-10
-
?
D-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
Pseudomonas putida pyruvate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
?
D-lactate + ubiquinone-10
-
Pseudomonas putida pyruvate + ubiquinol-10
-
?
L-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
Pseudomonas putida pyruvate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
?
L-lactate + ubiquinone-10
-
Pseudomonas putida pyruvate + ubiquinol-10
-
?
additional information the enzyme exhibits narrow substrate specificity. No activity with D-2-hydroxybutyrate, L-2-hydroxybutyrate, D-mandelate, L-mandelate, D-3-phenyllactate, D-glycerate, L-glycerate, DL-2-hydroxyisocaproate, and DL-2-hydroxyoctanoate, with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as an electron acceptor Pseudomonas putida ?
-
-

Subunits

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

Synonyms

Synonyms Comment Organism
Fe-S D-iLDH
-
Pseudomonas putida
lldE
-
Pseudomonas putida
NAD-independent D-lactate dehydrogenase
-
Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4
-
Ubiquinone-10 pH 7.4, 30°C Pseudomonas putida
8
-
D-glycerate pH 7.4, 30°C Pseudomonas putida
9.5
-
L-lactate pH 7.4, 30°C Pseudomonas putida
66
-
D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
120
-
D-lactate pH 7.4, 30°C Pseudomonas putida
128
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide pH 7.4, 30°C Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
FAD the enzyme contains a flavin adenine dinucleotide (FAD)-containing dehydrogenase domain. Compared to the intact enzyme, the FAD-containing dehydrogenase domain shows increased catalytic efficiency with cytochrome c as the electron acceptor, but it completely loses the ability to use coenzyme Q10. The 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
Fe-S center the enzyme contains an Fe-S oxidoreductase domain. The Fe-S oxidoreductase domain functions as an electron transfer component to facilitate the utilization of quinone as an electron acceptor by the enzyme (Fe-S D-iLDH), and it helps the enzyme associate with the cell membrane Pseudomonas putida

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.59
-
Ubiquinone-10 pH 7.4, 30°C Pseudomonas putida
6
-
D-glycerate pH 7.4, 30°C Pseudomonas putida
9
-
L-lactate pH 7.4, 30°C Pseudomonas putida
660
-
D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
2100
-
D-lactate pH 7.4, 30°C Pseudomonas putida
2700
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide pH 7.4, 30°C Pseudomonas putida