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Literature summary for 1.1.1.67 extracted from
- Structure-guided engineering of the coenzyme specificity of Pseudomonas fluorescens mannitol 2-dehydrogenase to enable efficient utilization of NAD(H) and NADP(H) (2008), FEBS Lett., 582, 233-237.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no activation of M2DH by bovine serum albumin | Pseudomonas fluorescens |
Application
| 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(Commentary)
| 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
| 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
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no inhibition of M2DH by bovine serum albumin | Pseudomonas fluorescens |
KM Value [mM]
| 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/ Products (Substrates)
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas fluorescens | O08355 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| mutants purified to apparent homogeneity | Pseudomonas fluorescens |
Substrates and Products (Substrate)
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| M2DH | - |
Pseudomonas fluorescens |
| mannitol 2-dehydrogenase | - |
Pseudomonas fluorescens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at, both reaction directions | Pseudomonas fluorescens |
Turnover Number [1/s]
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at, both reaction directions | Pseudomonas fluorescens |
Cofactor
| 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 | |
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