EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.13.33 | gene praI, functional recombinant expression in different strains of Pseudomonas putida replacing the endogenous gene pobA that encodes a PHBH enzyme | Paenibacillus sp. JJ-1b |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.14.13.33 | additional information | in Pseudomonas putida KT2440 strains engineered to convert lignin-related aromatic compounds to muconic acid (MA), PHBH activity is rate-limiting, as indicated by the accumulation of 4-HBA, which ultimately limits MA productivity. Replacement of PobA, the native Pseudomonas putida PHBH, with PraI, a PHBH from Paenibacillus sp. JJ-1b with a broader nicotinamide cofactor preference, can alleviate this bottleneck | Paenibacillus sp. JJ-1b |
1.14.13.33 | additional information | in Pseudomonas putida KT2440 strains engineered to convert lignin-related aromatic compounds to muconic acid (MA), PHBH activity is rate-limiting, as indicated by the accumulation of 4-hydroxybenzoate (4-HBA), which ultimately limits MA productivity. Replacement of PobA, the native Pseudomonas putida PHBH, with PraI, a PHBH from Paenibacillus sp. JJ-1b with a broader nicotinamide cofactor preference, can alleviate this bottleneck. Comparison of three Pseudomonas putida strains engineered to produce MA from p-coumarate (pCA), showing that expression of praI leads to lower 4-HBA accumulation and decreased NADP+/NADPH ratios relative to strains harboring pobA, indicative of a relieved 4-HBA bottleneck due to increased NADPH availability. In bioreactor cultivations, a strain exclusively expressing praI achieved a titer of 40 g/l MA at 100% molar yield and a productivity of 0.5 g/l/h | Pseudomonas putida |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.13.33 | 4-hydroxybenzoate + NADH + H+ + O2 | Pseudomonas putida | - |
3,4-dihydroxybenzoate + NAD+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADH + H+ + O2 | Paenibacillus sp. JJ-1b | - |
3,4-dihydroxybenzoate + NAD+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADPH + H+ + O2 | Pseudomonas putida | - |
3,4-dihydroxybenzoate + NADP+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADPH + H+ + O2 | Paenibacillus sp. JJ-1b | - |
3,4-dihydroxybenzoate + NADP+ + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.13.33 | Paenibacillus sp. JJ-1b | C4TP09 | - |
- |
1.14.13.33 | Pseudomonas putida | Q9R9T1 | - |
- |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.13.33 | 4-hydroxybenzoate + NADH + H+ + O2 | - |
Pseudomonas putida | 3,4-dihydroxybenzoate + NAD+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADH + H+ + O2 | - |
Paenibacillus sp. JJ-1b | 3,4-dihydroxybenzoate + NAD+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADPH + H+ + O2 | - |
Pseudomonas putida | 3,4-dihydroxybenzoate + NADP+ + H2O | - |
? | |
1.14.13.33 | 4-hydroxybenzoate + NADPH + H+ + O2 | - |
Paenibacillus sp. JJ-1b | 3,4-dihydroxybenzoate + NADP+ + H2O | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.13.33 | para-hydroxybenzoate-3-hydroxylases | - |
Pseudomonas putida |
1.14.13.33 | para-hydroxybenzoate-3-hydroxylases | - |
Paenibacillus sp. JJ-1b |
1.14.13.33 | PHBH | - |
Pseudomonas putida |
1.14.13.33 | PHBH | - |
Paenibacillus sp. JJ-1b |
1.14.13.33 | PobA | - |
Pseudomonas putida |
1.14.13.33 | PRAI | - |
Paenibacillus sp. JJ-1b |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.13.33 | additional information | enzyme PraI can use NADPH and also efficiently utilizes NADH at roughly half the catalytic efficiency | Paenibacillus sp. JJ-1b | |
1.14.13.33 | additional information | strict preference of NADPH for PobA | Pseudomonas putida | |
1.14.13.33 | NADH | - |
Paenibacillus sp. JJ-1b | |
1.14.13.33 | NADPH | - |
Pseudomonas putida | |
1.14.13.33 | NADPH | - |
Paenibacillus sp. JJ-1b |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.14.13.33 | additional information | the X-ray crystal structure of PraI is solved and reveals absolute conservation of the active site architecture to other PHBH structures despite their differing cofactor preferences | Paenibacillus sp. JJ-1b |
1.14.13.33 | physiological function | the transformation of 4-hydroxybenzoate (4-HBA) to protocatechuate (PCA) is catalyzed by flavoprotein oxygenases known as para-hydroxybenzoate-3-hydroxylases (PHBHs) | Pseudomonas putida |
1.14.13.33 | physiological function | the transformation of 4-hydroxybenzoate (4-HBA) to protocatechuate (PCA) is catalyzed by flavoprotein oxygenases known as para-hydroxybenzoate-3-hydroxylases (PHBHs) | Paenibacillus sp. JJ-1b |