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Literature summary extracted from
- Characterization of a novel immobilized biocatalyst obtained by matrix-assisted refolding of recombinant polyhydroxyoctanoate depolymerase from Pseudomonas putida KT2442 isolated from inclusion bodies (2011), J. Ind. Microbiol. Biotechnol., 38, 1203-1209.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.1.76 | overexpression of the His-tagged enzyme in Escherichia coli strain M15 in inclusion bodies | Pseudomonas putida |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 3.1.1.76 | additional information | PhaZ depolymerase is an intracellular depolymerase that is located in PHA granules | Pseudomonas putida | - |
- |
| 3.1.1.76 | soluble | - |
Pseudomonas putida | - |
- |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.1.76 | additional information | Pseudomonas putida | the enzyme is specific for medium-chain length PHA, i.e. mcl-PHA, with six or more carbon atoms per monomer. PhaZ depolymerase is an intracellular depolymerase that is located in PHA granules and hydrolyzes specifically mcl-PHAs containing aliphatic and aromatic monomers | ? | - |
? |  |
| 3.1.1.76 | additional information | Pseudomonas putida KT 2442 | the enzyme is specific for medium-chain length PHA, i.e. mcl-PHA, with six or more carbon atoms per monomer. PhaZ depolymerase is an intracellular depolymerase that is located in PHA granules and hydrolyzes specifically mcl-PHAs containing aliphatic and aromatic monomers | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.1.76 | Pseudomonas putida | - |
gene phaZ | - |
| 3.1.1.76 | Pseudomonas putida KT 2442 | - |
gene phaZ | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.1.1.76 | recombinant solubilized and refolded His-tagged enzyme from Escherichia coli strain M15 by nickel affinity chromatography. The refolded enzyme cannot be eluted with 1 M imidazole buffer, leading to an immobilized biocatalyst where PhaZ depolymerase is homogeneously distributed in the agarose support as shown by confocal scanning microscopy. Polyhydroxyoctanoate cannot be hydrolyzed by the immobilized biocatalyst, whereas the attached enzyme is active in the hydrolysis of 4-nitrophenyl alkanoate esters, which differed in their alkyl chain length | Pseudomonas putida |
Renatured (Commentary)
| EC Number | Renatured (Comment) | Organism |
|---|---|---|
| 3.1.1.76 | recombinant His-tagged enzyme from Escherichia coli strain M15 is solubilized in 8 M urea, immobilized on Ni2+-nitrilotriacetate-agarose matrix, and refolded by gradual removal of the chaotropic agent | Pseudomonas putida |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.1.76 | 4-nitrophenyl acetate + H2O | - |
Pseudomonas putida | 4-nitrophenol + acetate | - |
? | |
| 3.1.1.76 | 4-nitrophenyl acetate + H2O | - |
Pseudomonas putida KT 2442 | 4-nitrophenol + acetate | - |
? | |
| 3.1.1.76 | 4-nitrophenyl octanoate + H2O | - |
Pseudomonas putida | 4-nitrophenol + octanoate | - |
? | |
| 3.1.1.76 | 4-nitrophenyl octanoate + H2O | - |
Pseudomonas putida KT 2442 | 4-nitrophenol + octanoate | - |
? | |
| 3.1.1.76 | additional information | the enzyme is specific for medium-chain length PHA, i.e. mcl-PHA, with six or more carbon atoms per monomer. PhaZ depolymerase is an intracellular depolymerase that is located in PHA granules and hydrolyzes specifically mcl-PHAs containing aliphatic and aromatic monomers | Pseudomonas putida | ? | - |
? | |
| 3.1.1.76 | additional information | the enzyme is specific for medium-chain length PHA, i.e. mcl-PHA, with six or more carbon atoms per monomer. PhaZ depolymerase is an intracellular depolymerase that is located in PHA granules and hydrolyzes specifically mcl-PHAs containing aliphatic and aromatic monomers | Pseudomonas putida KT 2442 | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.1.76 | PHA depolymerase | - |
Pseudomonas putida |
| 3.1.1.76 | PhaZ | - |
Pseudomonas putida |
| 3.1.1.76 | polyhydroxyalkanoate depolymerase | - |
Pseudomonas putida |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.76 | 37 | - |
assay at | Pseudomonas putida |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.76 | 40 | - |
the enzyme immobilized on agarose maintains 100% activity up to 40°C, but the activity quickly diminishes at higher temperatures | Pseudomonas putida |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.76 | 8 | - |
assay at | Pseudomonas putida |
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