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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, e00342-17 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.5.12

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
C540S [Fe-S]-cluster-to-protein ratio is reduced to 98% of wild-type Pseudomonas putida
C540S/594S/C751S/C752S [Fe-S]-cluster-to-protein ratio is reduced to 27.8% of wild-type Pseudomonas putida
C540S/C543S/C594S/C751S/C752S/C540S/C594S [Fe-S]-cluster-to-protein ratio is reduced to 1.1% of wild-type Pseudomonas putida
C540S/C594S [Fe-S]-cluster-to-protein ratio is reduced to 60.9% of wild-type Pseudomonas putida
C540S/C594S/C751S/C752S/C540S/C594S [Fe-S]-cluster-to-protein ratio is reduced to 9% of wild-type Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
antimycin A inhibits electron transfer from quinone to cytochrome c Pseudomonas putida
Ba2+ 5 mM, partial inhibition; 5 mM, partial inhibition of activity Pseudomonas putida
Ca2+ 5 mM, partial inhibition; 5 mM, partial inhibition of activity Pseudomonas putida
Cd2+ 5 mM, complete inhibition; 5 mM, complete inhibition of activity Pseudomonas putida
Co2+ 5 mM, complete inhibition; 5 mM, complete inhibition of activity Pseudomonas putida
Cu2+ 5 mM, complete inhibition; 5 mM, complete inhibition of activity Pseudomonas putida
EDTA 5 mM, partial inhibition; 5 mM, partial inhibition of activity Pseudomonas putida
Fe2+ 5 mM, complete inhibition; 5 mM, complete inhibition of activity Pseudomonas putida
Mg2+ 5 mM, partial inhibition; 5 mM, partial inhibition of activity Pseudomonas putida
Mn2+ 5 mM, complete inhibition; 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; 5 mM, partial inhibition of activity Pseudomonas putida
Zn2+ 5 mM, complete inhibition; 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
0.057
-
 (R)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
0.1
-
 D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
0.1
-
 D-2-hydroxybutyrate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
1.04
-
 L-lactate pH 7.4, 30°C Pseudomonas putida
1.04
-
 (S)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
1.38
-
 D-glycerate pH 7.4, 30°C Pseudomonas putida
1.38
-
 D-glycerate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida

Localization

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

Metals/Ions

Metals/Ions Comment Organism Structure
K+ 5 mM, slightly positive effect Pseudomonas putida
K+ exhibits a slightly positive effect activity 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
purified as a His-tagged protein. The two domains of the enzyme are heterologously expressed and purified separately Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-lactate + 2 ferricytochrome c quinone is the preferred electron acceptor of the enzyme Pseudomonas putida pyruvate + 2 ferrocytochrome c
-
 ?
(R)-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide quinone is the preferred electron acceptor of the enzyme Pseudomonas putida pyruvate + ?
-
 ?
(R)-lactate + coenzyme Q10 quinone is the preferred electron acceptor of the enzyme Pseudomonas putida pyruvate + reduced coenzyme Q10
-
 ?
(S)-lactate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide quinone is the preferred electron acceptor of the enzyme Pseudomonas putida pyruvate + ?
-
 ?
D-2-hydroxybutyrate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida 2-oxobutanoate + ?
-
 ?
D-2-hydroxybutyrate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida ? + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
D-2-hydroxybutyrate + ubiquinone-10
-
 Pseudomonas putida ? + ubiquinol-10
-
 ?
D-glycerate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida ?
-
 ?
D-glycerate + 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 Pseudomonas putida ? + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
 ?
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
pp4737
-
 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
8
-
 D-glycerate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
9.5
-
 L-lactate pH 7.4, 30°C Pseudomonas putida
9.5
-
 (S)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
66
-
 D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
66
-
 D-2-hydroxybutyrate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
120
-
 D-lactate pH 7.4, 30°C Pseudomonas putida
120
-
 (R)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide 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
FAD the enzyme contains one FAD molecule 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
Fe-S cluster the enzyme contains an Fe-S oxidoreductase domain that functions as an electron transfer component to facilitate the utilization of quinone as an electron acceptor. It helps the enzyme to 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
2.7
-
 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide pH 7.4, 30°C Pseudomonas putida
6
-
 D-glycerate pH 7.4, 30°C Pseudomonas putida
6
-
 D-glycerate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
9
-
 L-lactate pH 7.4, 30°C Pseudomonas putida
9
-
 (S)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
660
-
 D-2-hydroxybutyrate pH 7.4, 30°C Pseudomonas putida
660
-
 D-2-hydroxybutyrate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida
2100
-
 D-lactate pH 7.4, 30°C Pseudomonas putida
2100
-
 (R)-lactate pH 7.4, 30°C, acceptor: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Pseudomonas putida