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Literature summary for 4.2.1.119 extracted from
- Contribution of the distal pocket residue to the acyl-chain-length specificity of (R)-specific enoyl-coenzyme A hydratases from Pseudomonas spp. (2015), Appl. Environ. Microbiol., 81, 8076-8083 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene phaJ1, sequence comparison, recombinant expression of wild-type and mutant enzymes in Escherichia coli strains DH5alpha or JM109, and LS5218, overexpression of the enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas aeruginosa |
| gene phaJ1, sequence comparison, recombinant expression of wild-type and mutant enzymes in Escherichia coli strains DH5alpha or JM109, and LS5218, overexpression of the enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| homology modeling of strcuture. In the acyl-chain binding pocket, the amino acid at position 72 is the only difference between the two structures of Pseudomonas aeruginosa and Pseudomonas putida isoforms | Pseudomonas aeruginosa |
| homology modeling of strcuture. In the acyl-chain binding pocket, the amino acid at position 72 is the only difference between the two structures of Pseudomonas aeruginosa and Pseudomonas putida isoforms | Pseudomonas putida |
| purified recombinant enzyme PhaJ1Pa, sitting drop vapor diffusion, mixing of 10 mg/ml protein in 20 mM Tris-HCl, pH 7.5, with mother liquor containing 15 to 20% w/v PEG 3350, 20% v/v glycerol, and 0.1 M bis-Tris, pH 6.0-6.5, X-ray diffraction structure determination and analysis at 1.7 A resolution | Pseudomonas aeruginosa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| I72V | site-directed mutagenesis, the mutant has an increased preference for enoyl-CoAs with longer chain lengths compared to wild-type broadening the substrate specificity range, PHA accumulation in recombinant Escherichia coli LS5218 harboring parental or Val72/Ile72 mutant genes of phaJ1, overview | Pseudomonas aeruginosa |
| additional information | construction of three chimeric PhaJ1 enzymes, composed from Pseudomonas aeruginosa and Pseudomonas putida isoforms. All chimera show significant hydratase activity, and their substrate preferences is within the range exhibited by the parental PhaJ1 enzymes | Pseudomonas aeruginosa |
| additional information | construction of three chimeric PhaJ1 enzymes, composed from Pseudomonas aeruginosa and Pseudomonas putida isoforms. All chimera show significant hydratase activity, and their substrate preferences is within the range exhibited by the parental PhaJ1 enzymes | Pseudomonas putida |
| additional information | construction of three chimeric PhaJ1 enzymes, composed from the enzymes from Pseudomonas putida and Pseudomonas aeruginosa, PhaJ1Pp and PhaJ1Pa. All mutants show significant hydratase activity, and their substrate preferences are within the range exhibited by the parental PhaJ1 enzymes | Pseudomonas aeruginosa |
| additional information | construction of three chimeric PhaJ1 enzymes, composed from the enzymes from Pseudomonas putida and Pseudomonas aeruginosa, PhaJ1Pp and PhaJ1Pa. All mutants show significant hydratase activity, and their substrate preferences are within the range exhibited by the parental PhaJ1 enzymes | Pseudomonas putida |
| V72I | site-directed mutagenesis, the mutant has an increased preference for enoyl-coenzyme A (CoA) elements with shorter chain lengths compared to wild-type narrowing the substrate specificity range, PHA accumulation in recombinant Escherichia coli LS5218 harboring parental or Val72/Ile72 mutant genes of phaJ1, overview | Pseudomonas putida |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (2E)-2-enoyl-CoA + H2O | Pseudomonas aeruginosa | - |
(3R)-3-hydroxyacyl-CoA | - |
r |  |
| (2E)-2-enoyl-CoA + H2O | Pseudomonas putida | - |
(3R)-3-hydroxyacyl-CoA | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | Q9LBK2 | - |
- |
| Pseudomonas putida | A0A177YY42 | - |
- |
| Pseudomonas putida | E9P3X9 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Pseudomonas aeruginosa |
| recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Pseudomonas putida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (2E)-2-enoyl-CoA + H2O | - |
Pseudomonas aeruginosa | (3R)-3-hydroxyacyl-CoA | - |
r | |
| (2E)-2-enoyl-CoA + H2O | - |
Pseudomonas putida | (3R)-3-hydroxyacyl-CoA | - |
r | |
| (2E)-crotonyl-CoA + H2O | - |
Pseudomonas aeruginosa | (3R)-3-hydroxybutanoyl-CoA | - |
r | |
| (2E)-crotonyl-CoA + H2O | - |
Pseudomonas putida | (3R)-3-hydroxybutanoyl-CoA | - |
r | |
| (2E)-oct-2-enoyl-CoA + H2O | - |
Pseudomonas aeruginosa | (R)-3-hydroxyoctanoyl-CoA | - |
r | |
| (2E)-oct-2-enoyl-CoA + H2O | - |
Pseudomonas putida | (R)-3-hydroxyoctanoyl-CoA | - |
r | |
| additional information | chain-length specificity of PhaJ1 is determined mainly by the bulkiness of the amino acid residue at position 72, but other factors, such as structural fluctuations, also affect specificity | Pseudomonas aeruginosa | ? | - |
? | |
| additional information | chain-length specificity of PhaJ1 is determined mainly by the bulkiness of the amino acid residue at position 72, but other factors, such as structural fluctuations, also affect specificity | Pseudomonas putida | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| (R)-specific enoyl-coenzyme A hydratase | - |
Pseudomonas aeruginosa |
| (R)-specific enoyl-coenzyme A hydratase | - |
Pseudomonas putida |
| PhaJ1 | - |
Pseudomonas aeruginosa |
| PhaJ1 | - |
Pseudomonas putida |
| PhaJ1Pa | - |
Pseudomonas aeruginosa |
| PhaJ1Pp | - |
Pseudomonas putida |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Pseudomonas aeruginosa |
| 30 | - |
assay at | Pseudomonas putida |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Pseudomonas aeruginosa |
| 8 | - |
assay at | Pseudomonas putida |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | contribution of the distal pocket residue to the acyl-chain-length specificity of (R)-specific enoyl-coenzyme A hydratases from Pseudomonas spp., enzyme structure homology modeling, structure comparisons of the enzymes from Pseudomonas putida and Pseudomonas aeruginosa, PhaJ1Pp and PhaJ1Pa, overview | Pseudomonas putida |
| additional information | contribution of the distal pocket residue to the acyl-chain-length specificity of (R)-specific enoyl-coenzyme A hydratases from Pseudomonas spp., enzyme structure homology modeling, structure comparisons of the enzymes from Pseudomonas putida and Pseudomonas aeruginosa, PhaJ1Pp and PhaJ1Pa, overview. Active site and acyl-chain-binding pocket structure | Pseudomonas aeruginosa |
| physiological function | (R)-specific enoyl-coenzyme A (enoyl-CoA) hydratases (PhaJs) are capable of supplying monomers from fatty acid beta-oxidation to polyhydroxyalkanoate (PHA) biosynthesis. PhaJ1Pp from Pseudomonas putida shows a broader substrate specificity | Pseudomonas aeruginosa |
| physiological function | (R)-specific enoyl-coenzyme A (enoyl-CoA) hydratases (PhaJs) are capable of supplying monomers from fatty acid beta-oxidation to polyhydroxyalkanoate (PHA) biosynthesis. PhaJ1Pp from Pseudomonas putida shows a broader substrate specificity | Pseudomonas putida |
| physiological function | comparison of the enzymes from Pseudomonas putida with residue 72Val resulting in increased preference for enoyl-coenzyme A substrates with shorter chain lengths and Pseudomonas aeruginosa with residue 72Ile resulting in an increased preference for enoyl-CoAs with longer chain lengths | Pseudomonas putida |
| physiological function | comparison of the enzymes from Pseudomonas putida with residue Val72 resulting in increased preference for enoyl-coenzyme A substrates with shorter chain lengths and Pseudomonas aeruginosa with residue Ile72 resulting in an increased preference for enoyl-CoAs with longer chain lengths | Pseudomonas aeruginosa |
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Release 2023.1 (January 2023)
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