Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21(DE3) | Streptococcus pneumoniae |
Crystallization (Comment) | Organism |
---|---|
micro-batch and hanging-drop vapor-diffusion methods. The crystal structure is determined at 1.69 A resolution. The crystal structure of SpIlvC contains an asymmetric dimer in which one subunit is in apo-form and the other in NADP(H) and Mg2+-bound form | Streptococcus pneumoniae |
Protein Variants | Comment | Organism |
---|---|---|
D191G | crystal structure of the mutants (R49E, D83G, D191G and E195S) reveals local conformational changes only in the NADP(H) binding site. Mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
D191K | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
D83G | crystal structure of the mutants (R49E, D83G, D191G and E195S) reveals local conformational changes only in the NADP(H) binding site. Mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
E195A | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
E195K | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
E195S | crystal structure of the mutants (R49E, D83G, D191G and E195S) reveals local conformational changes only in the NADP(H) binding site. Mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
R49A | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
R49E | crystal structure of the mutants (R49E, D83G, D191G and E195S) reveals local conformational changes only in the NADP(H) binding site. Mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
R49G | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
S53A | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
S53G | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
S53K | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
S53T | mutant enzyme exhibits significantly reduced activities compared to wild-type enzyme | Streptococcus pneumoniae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
NADPH | wild-type enzyme, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.199 | - |
NADPH | mutant enzyme S53G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.33 | - |
NADPH | mutant enzyme D83G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.345 | - |
NADPH | mutant enzyme S53T, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.355 | - |
NADPH | mutant enzyme D191G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.395 | - |
NADPH | mutant enzyme R49G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.47 | 1 | NADPH | mutant enzyme S53K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.472 | - |
NADPH | mutant enzyme R49E, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.588 | - |
NADPH | mutant enzyme R49A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.677 | - |
NADPH | mutant enzyme S53A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.732 | - |
NADPH | mutant enzyme E195A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.895 | - |
NADPH | mutant enzyme E195K, pH 8.0, 30°C | Streptococcus pneumoniae | |
1.036 | - |
NADPH | mutant enzyme D191K, pH 8.0, 30°C | Streptococcus pneumoniae | |
1.467 | - |
NADPH | mutant enzyme E195S, pH 8.0, 30°C | Streptococcus pneumoniae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pneumoniae | Q04M32 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Streptococcus pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2S)-2-acetolactate + NADPH + H+ | - |
Streptococcus pneumoniae | (R)-2,3-dihydroxyisovalerate + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | the enzyme retains dimeric arrangement in solution | Streptococcus pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
ilvC | - |
Streptococcus pneumoniae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
NADPH | wild-type enzyme, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.11 | - |
NADPH | mutant enzyme S53G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.12 | - |
NADPH | mutant enzyme D191K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.12 | - |
NADPH | mutant enzyme E195A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.12 | - |
NADPH | mutant enzyme S53T, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.13 | - |
NADPH | mutant enzyme D83G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.14 | - |
NADPH | mutant enzyme S53K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.15 | - |
NADPH | mutant enzyme D191G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.16 | - |
NADPH | mutant enzyme R49G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.17 | - |
NADPH | mutant enzyme S53A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.18 | - |
NADPH | mutant enzyme E195K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.18 | - |
NADPH | mutant enzyme R49A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.18 | - |
NADPH | mutant enzyme R49E, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.25 | - |
NADPH | mutant enzyme E195S, pH 8.0, 30°C | Streptococcus pneumoniae |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme catalyzes the second step in the BCAA biosynthetic pathway | Streptococcus pneumoniae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
NADPH | mutant enzyme E195A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.17 | - |
NADPH | mutant enzyme E195S, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.2 | - |
NADPH | mutant enzyme E195K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.21 | - |
NADPH | mutant enzyme D191K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.25 | - |
NADPH | mutant enzyme S53A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.3 | - |
NADPH | mutant enzyme R49A, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.3 | - |
NADPH | mutant enzyme S53K, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.34 | - |
NADPH | mutant enzyme S53T, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.38 | - |
NADPH | mutant enzyme R49E, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.39 | - |
NADPH | mutant enzyme D83G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.42 | - |
NADPH | mutant enzyme D191G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.43 | - |
NADPH | mutant enzyme R49G, pH 8.0, 30°C | Streptococcus pneumoniae | |
0.55 | - |
NADPH | mutant enzyme S53G, pH 8.0, 30°C | Streptococcus pneumoniae | |
1.4 | - |
NADPH | wild-type enzyme, pH 8.0, 30°C | Streptococcus pneumoniae |