Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | no activation of M2DH by bovine serum albumin | Pseudomonas fluorescens |
Application | Comment | Organism |
---|---|---|
industry | redox balancing between the intracellular NADP(H) and NAD(H) based on NAD(P)(H)-dependent interconversion of mannitol and fructose by M2DH may be a useful strategy of metabolic engineering | Pseudomonas fluorescens |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Pseudomonas fluorescens |
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
D69A | site-directed mutagenesis, the mutant shows an altered cofactor specificity compared to the wild-type enzyme, which is switched to NADP(H), EC 1.1.1.138, NADP(H) is equally utilized as NAD(H) | Pseudomonas fluorescens |
D69A | utilizes NAD(H) and NADP(H) with similar catalytic efficiencies. Uses NADP(H) almost as well as wild-type enzyme uses NAD(H) | Pseudomonas fluorescens |
E68K | site-directed mutagenesis, the mutant shows an altered cofactor specificity compared to the wild-type enzyme, which is switched to NADP(H), EC 1.1.1.138, NADP(H) is preferred by 10fold over NAD(H) | Pseudomonas fluorescens |
E68K/D69A | shows about a 10fold preference for NADP(H) over NAD(H), accompanied by a small decrease in catalytic efficiency for NAD(H)-dependent reactions as compared to wild-type enzyme | Pseudomonas fluorescens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no inhibition of M2DH by bovine serum albumin | Pseudomonas fluorescens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis, recombinant wild-type and mutant enzymes, overview | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-mannitol + NAD+ | Pseudomonas fluorescens | wild-type enzyme | D-fructose + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | O08355 | - |
- |
Purification (Comment) | Organism |
---|---|
mutants purified to apparent homogeneity | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-mannitol + NAD(P)+ | - |
Pseudomonas fluorescens | D-fructose + NAD(P)H + H+ | - |
r | |
D-mannitol + NAD+ | wild-type enzyme | Pseudomonas fluorescens | D-fructose + NADH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
M2DH | - |
Pseudomonas fluorescens |
mannitol 2-dehydrogenase | - |
Pseudomonas fluorescens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at, both reaction directions | Pseudomonas fluorescens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | - |
NAD(P)H | wild-type | Pseudomonas fluorescens | |
4.6 | - |
NADH | mutant D69A/D69A | Pseudomonas fluorescens | |
12 | - |
NAD(P)H | mutant D69A/D69A | Pseudomonas fluorescens | |
15 | - |
NADH | mutant D69A | Pseudomonas fluorescens | |
24 | - |
NAD(P)H | mutant D69A | Pseudomonas fluorescens | |
26 | - |
NADH | wild-type | Pseudomonas fluorescens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at, both reaction directions | Pseudomonas fluorescens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas fluorescens | |
NAD+ | binding structure, the carboxylate group of Asp69 forms a bifurcated hydrogen bond with the 2' and 3' hydroxyl groups of the adenosine of NAD+ and contributes to the 400fold preference of the enzyme for NAD+ as compared to NADP+, overview | Pseudomonas fluorescens | |
NADH | - |
Pseudomonas fluorescens | |
NADP+ | 400fold preference of the enzyme for NAD+ as compared to NADP+ | Pseudomonas fluorescens |