Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 | Pseudomonas mevalonii |
Crystallization (Comment) | Organism |
---|---|
wild-type and mutants, H381A, H381N, H381K, H381Q, 3 A resolution | Pseudomonas mevalonii |
Protein Variants | Comment | Organism |
---|---|---|
H381A | 6% of wild-type activity | Pseudomonas mevalonii |
H381K | 45% of wild-type activity at pH 11 | Pseudomonas mevalonii |
H381N | 16% of wild-type activity | Pseudomonas mevalonii |
H381Q | 23% of wild-type activity | Pseudomonas mevalonii |
General Stability | Organism |
---|---|
diethyl polycarbonate causes loss of activity | Pseudomonas mevalonii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
CoASH | H381A mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.05 | - |
CoASH | wild-type enzyme, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.11 | - |
CoASH | H381Q mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.14 | - |
NAD+ | H381A mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.2 | - |
CoASH | H381N mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.22 | - |
NAD+ | wild-type enzyme, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.27 | - |
(R,S)-mevalonate | H381A mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.45 | - |
(R,S)-mevalonate | wild-type enzyme, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.5 | - |
CoASH | H381K mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.62 | - |
NAD+ | H381Q mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.71 | - |
(R,S)-mevalonate | H381Q mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.75 | - |
(R,S)-mevalonate | H381K mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
0.81 | - |
NAD+ | H381N mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
1.23 | - |
NAD+ | H381K mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
3.7 | - |
(R,S)-mevalonate | H381N mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas mevalonii | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas mevalonii |
Renatured (Comment) | Organism |
---|---|
after inactivation with diethyl polycarbonate addition of hydroxylamine restores 50% of activity | Pseudomonas mevalonii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.011 | - |
H381A-mutant, mevalonate acylation | Pseudomonas mevalonii |
0.029 | - |
H381N-mutant, mevalonate acylation | Pseudomonas mevalonii |
0.041 | - |
H381Q-mutant, mevalonate acylation | Pseudomonas mevalonii |
0.091 | - |
H381K-mutant, mevalonate acylation | Pseudomonas mevalonii |
18 | - |
mevalonate acylation pH 8.1 | Pseudomonas mevalonii |
20.3 | - |
mevalonate acylation pH 11 | Pseudomonas mevalonii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-mevalonate + CoA + 2 NAD+ | - |
Pseudomonas mevalonii | (S)-3-hydroxy-3-methylglutaryl-CoA + 2 NADH + 2 H+ | - |
? | |
3-hydroxy-3-methylglutaryl-CoA + NADH + H+ | - |
Pseudomonas mevalonii | mevalonate + CoA + NAD+ | - |
? |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
oxidative acylation of mevalonate | Pseudomonas mevalonii |
9.5 | - |
H381N mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii |
10 | - |
H381A mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii |
10.5 | - |
H381Q mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii |
11 | - |
H381K mutant, oxidative acylation of mevalonate | Pseudomonas mevalonii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas mevalonii | |
NADH | reverse reaction | Pseudomonas mevalonii |