Cloned (Comment) | Organism |
---|---|
gene nahI, the naphthalene catabolic genes (nah) of NAH7 are organized into two operons on a 83 kilobase plasmid, recombinant expression of soluble N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
enzyme structure analysis by dynamic light scattering, small-angle X-ray scattering experiments and circular dichroism spectroscopy | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Pseudomonas putida | |
0.0013 | - |
2-hydroxymuconate-6-semialdehyde | pH 8.5, 25°C, recombinant enzyme | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
184000 | - |
recombinant enzyme, gel filtration | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxymuconate-6-semialdehyde + NAD+ + H2O | Pseudomonas putida | - |
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate + NADH + 2 H+ | - |
? | |
2-hydroxymuconate-6-semialdehyde + NAD+ + H2O | Pseudomonas putida G7 | - |
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate + NADH + 2 H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | Q1XGK8 | - |
- |
Pseudomonas putida G7 | Q1XGK8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant soluble N-terminally His6-tagged enzyme from Escherichia coli by nickel affinity chromatography, tag cleavage by TEV protease, and another nickel affinity chromatography step, followed by gel filtration and ultrafiltration | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxymuconate-6-semialdehyde + NAD+ + H2O | - |
Pseudomonas putida | (2Z,4E)-2-hydroxyhexa-2,4-dienedioate + NADH + 2 H+ | - |
? | |
2-hydroxymuconate-6-semialdehyde + NAD+ + H2O | - |
Pseudomonas putida G7 | (2Z,4E)-2-hydroxyhexa-2,4-dienedioate + NADH + 2 H+ | - |
? | |
additional information | enzyme NahI is highly specific for its biological substrate, 2-hydroxymuconate semialdehyde, no activity with salicylaldehyde, another intermediate in the naphthalene-degradation pathway | Pseudomonas putida | ? | - |
? | |
additional information | enzyme NahI is highly specific for its biological substrate, 2-hydroxymuconate semialdehyde, no activity with salicylaldehyde, another intermediate in the naphthalene-degradation pathway | Pseudomonas putida G7 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | enzyme protein structure analysis by circular dichroism spectroscopy, and three-dimensional structure model, overview | Pseudomonas putida |
tetramer | x * 51712, recombinant detagged enzyme, mass spectrometry, x * 55000, recombinant N-terminally His-tagged enzyme, SDS-PAGE | Pseudomonas putida |
Synonyms | Comment | Organism |
---|---|---|
2-hydroxymuconate semialdehyde dehydrogenase | - |
Pseudomonas putida |
NahI | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
2-hydroxymuconate-6-semialdehyde | pH 8.5, 25°C, recombinant enzyme | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pseudomonas putida |
General Information | Comment | Organism |
---|---|---|
evolution | the naphthalene catabolic genes (nah) of NAH7 are organized into two operons. The nah operon (nahAaAbAcAdBFCED) encodes the upper pathway enzymes involved in the conversion of naphthalene to salicylate. On the other hand, the sal operon (nahGTHINLOMKJ) codes for the lower pathway enzymes involved in the conversion of salicylate to pyruvate and acetaldehyde. The lower pathway starts with the oxidation of salicylate to catechol by salicylate hydroxylase (NahG), which is extradiol-cleaved by catechol-2,3-dioxygenase (NahH) and further transformed to pyruvate and acetyl-CoA by the remaining meta-cleavage pathway gene products, NahI, NahJ, NahK, NahN, NahL, NahM, and NahO. Enzyme NahI is further classified into the ALDH8 family together with different 2-hydroxymuconate semialdehyde dehydrogenases | Pseudomonas putida |
physiological function | the enzyme is involved in the degradation of intermediate 2-hydroxymuconate 6-semialdehyde in the naphthalene-degradation pathway | Pseudomonas putida |