Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 | Pseudomonas mevalonii |
Protein Variants | Comment | Organism |
---|---|---|
D146A | 6670-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146A/L148K | 72200-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146A/L148R | 83300-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146A/Q147K | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
D146A/T192K | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
D146A/T192R | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
D146G | 1170-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146G/L148K | 55600-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146G/L148R | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
D146G/T192K | 3170-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146G/T192R | 4500-fold more specific for NADP+ than wild-type | Pseudomonas mevalonii |
D146N | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
D146S | no activity with NADP+ as cofactor | Pseudomonas mevalonii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.21 | - |
NAD+ | wild-type enzyme, oxidative acylation of mevalonate | Pseudomonas mevalonii | |
52 | - |
NADP+ | wild-type enzyme, oxidative acylation of mevalonate | Pseudomonas mevalonii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas mevalonii | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-mevalonate + CoA + NADP+ | - |
Pseudomonas mevalonii | 3-hydroxy-3-methylglutaryl-CoA + NADPH + H+ | - |
r |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | very detailed kinetic studies of mutants concerning NAD+/NADP+-affinities, the best specificity improvement is achieved by the D146A/L148R-mutant | Pseudomonas mevalonii | |
NAD+ | - |
Pseudomonas mevalonii | |
NADH | reverse reaction | Pseudomonas mevalonii | |
NADP+ | NAD+ is 600000fold more efficient than NADP+ in wild-type enzyme | Pseudomonas mevalonii |