Cloned (Comment) | Organism |
---|---|
expression of mutant enzymes Y201F and Y385F in Escherichia coli | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
Y201F | less than 6% of the activity of the wild-type enzyme. Reduction of FAD by NADPH is slower by 10fold, when the mutant enzyme-4-hydroxybenzoate complex reacts with oxygen, a long-lived flavin-C(4a)-hydroperoxide is observed, which slowly eliminates H2O2 with very little hydroxylation | Pseudomonas aeruginosa |
Y385F | less than 6% of the activity of the wild-type enzyme. Reduction of FAD by NADPH is slower by 100fold, the mutant enzyme reacts with oxygen to form 25% oxidized enzyme and 75% flavin hydroperoxide, which successfully hydroxylates the substrate. The mutant also hydroxylates the product 3,4-dihydroxybenzoate to form gallic acid | Pseudomonas aeruginosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybenzoate + NADPH + O2 | - |
Pseudomonas aeruginosa | protocatechuate + NADP+ + H2O | - |
? | |
additional information | mutant enzyme Y385F hydroxylates 3,4-dihydroxybenzoate to form gallic acid | Pseudomonas aeruginosa | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.36 | - |
4-hydroxybenzoate | mutant enzyme Y385F | Pseudomonas aeruginosa | |
0.42 | - |
4-hydroxybenzoate | mutant enzyme Y201F | Pseudomonas aeruginosa | |
5.7 | - |
4-hydroxybenzoate | wild-type enzyme | Pseudomonas aeruginosa |