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Literature summary for 1.2.1.46 extracted from
- Expression, purification, and characterization of formaldehyde dehydrogenase from Pseudomonas aeruginosa (2013), Protein Expr. Purif., 92, 208-213 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| additional information | the enzyme from Pseudomonas aeruginosa does not depend on glutathione for catalysis. The enzyme requires only NAD+ as the electron carrier, and addition of glutathione has no effect on the catalytic activity, indicating glutathione-independent catalysis | Pseudomonas aeruginosa |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)/pG-TF2. 50% of the recombinant proteins are localized in the supernatant fraction of cell-free extract, suggesting that coexpression with chaperones GroES, GroEL, and Tig has significantly improved the solubilization of Pseudomonas aeruginosa FDH in Escherichia coli | Pseudomonas aeruginosa |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant His-tagged enzyme, sitting drop vapor diffusion method, mixing of 0.001 ml of 15 mg/ml protein in 10 mM Tris-HCl. pH 8.0, and 1 mM DTT with 0.001 ml of reservoir solution containing 0.1 M Bis-Tris, pH 5.5, and 1.8 M ammonium sulfate, at 18°C, X-ray diffraction structure determination and analysis at 2.7 A resolution, molecular replacement method, and modelling | Pseudomonas aeruginosa |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | bi-substrate kinetics modelling | Pseudomonas aeruginosa |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Zn2+ | required | Pseudomonas aeruginosa |  |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 150000 | - |
recombinant His-tagged enzyme, gel filtration | Pseudomonas aeruginosa |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| formaldehyde + NAD+ + H2O | Pseudomonas aeruginosa | - |
formate + NADH + 2 H+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme 94.1fold from Escherichia coli strain BL21(DE3)/pG-TF2 by nickel affinity chromatography dialysis, anion exchange chromatography, and gelfiltration | Pseudomonas aeruginosa |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| formaldehyde + NAD+ + H2O = formate + NADH + H+ | the catalytic reaction proceeds by a Ping Pong mechanism as designated by the Michaelis-Menten model of bi-substrate enzymatic kinetics | Pseudomonas aeruginosa |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 9.87 | - |
purified recombinant His-tagged enzyme, pH 7.5, 37°C | Pseudomonas aeruginosa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| formaldehyde + NAD+ + H2O | - |
Pseudomonas aeruginosa | formate + NADH + 2 H+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotetramer | 4 * 44000, recombinant His-tagged enzyme, SDS-PAGE, 4 * 43252, recombinant His-tagged enzyme, mass spectrometry | Pseudomonas aeruginosa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| FDH | - |
Pseudomonas aeruginosa |
| glutathione-independent FDH | - |
Pseudomonas aeruginosa |
| glutathione-independent formaldehyde dehydrogenase | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 37 | assay at | Pseudomonas aeruginosa |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Pseudomonas aeruginosa |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | - |
Pseudomonas aeruginosa | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme is a member of zinc-containing medium-chain alcohol dehydrogenase family | Pseudomonas aeruginosa |
| physiological function | the enzyme oxidizes toxic formaldehyde to less active formate using NAD+ as a cofactor. The enzyme requires only NAD+ as the electron carrier, and addition of glutathione has no effect on the catalytic activity, indicating glutathione-independent catalysis | Pseudomonas aeruginosa |
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