Cloned (Comment) | Organism |
---|---|
recombinant His-tagged enzyme expression in Escherichia coli strain Rosetta 2 (DE3) as soluble protein | Pseudomonas sp. |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, sitting drop vapour diffusion metod, mixing of 150 nl of 10.5 mg/ml protein in 25 mM Tris chloride, 137 mM NaCl, and 3 mM KCl, pH 8.0, with 150 nl of reservoir solution containing 0.066 M calcium acetate, 18% w/v PEG MME 5000, 0.1 M Tris chloride, pH 7.6, and equilibration against 0.5 ml of reservoir solution, at 20°C, method optimization, X-ray diffraction structure determination analysis at 2.07 A resolution, molecular replacement method, structure modelling using the structure of a Chromobacterium violaceum omega transaminase monomer (PDB ID 4a6t) as the search model | Pseudomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
gabaculine | a transaminase suicide inhibitor | Pseudomonas sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
putrescine + 2-oxoglutarate | Pseudomonas sp. | - |
1-pyrroline + L-glutamate + H2O | - |
? | |
putrescine + 2-oxoglutarate | Pseudomonas sp. AAC | - |
1-pyrroline + L-glutamate + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Pseudomonas sp. AAC | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant soluble His-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) microfluidized cell extract by nickel affinity chromatography, gel filtration, and ultrafiltration | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
12-aminododecanoic acid + 2-oxoglutarate | - |
Pseudomonas sp. | ? | - |
? | |
12-aminododecanoic acid + 2-oxoglutarate | - |
Pseudomonas sp. AAC | ? | - |
? | |
additional information | the activity of KES24511 is determined using an alanine dehydrogenase enzyme-coupled assay, the purified protein exhibited activity with two substrates, putrescine and 12-aminododecanoic acid, with pyruvate as the co-substrate for transamination. The transaminase reaction co-product (L-alanine) is converted in situ by the dehydrogenase in an NAD+-dependent oxidation, and nicotinamide conversion is spectrophotometrically monitored by the change in UV absorbance at 340 nm | Pseudomonas sp. | ? | - |
- |
|
additional information | the activity of KES24511 is determined using an alanine dehydrogenase enzyme-coupled assay, the purified protein exhibited activity with two substrates, putrescine and 12-aminododecanoic acid, with pyruvate as the co-substrate for transamination. The transaminase reaction co-product (L-alanine) is converted in situ by the dehydrogenase in an NAD+-dependent oxidation, and nicotinamide conversion is spectrophotometrically monitored by the change in UV absorbance at 340 nm | Pseudomonas sp. AAC | ? | - |
- |
|
putrescine + 2-oxoglutarate | - |
Pseudomonas sp. | 1-pyrroline + L-glutamate + H2O | - |
? | |
putrescine + 2-oxoglutarate | overall reaction | Pseudomonas sp. | 1-pyrroline + L-glutamate + H2O | - |
? | |
putrescine + 2-oxoglutarate | - |
Pseudomonas sp. AAC | 1-pyrroline + L-glutamate + H2O | - |
? | |
putrescine + 2-oxoglutarate | overall reaction | Pseudomonas sp. AAC | 1-pyrroline + L-glutamate + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
FG99_07980 | - |
Pseudomonas sp. |
KES24511 | - |
Pseudomonas sp. |
putrescine aminotransferase | - |
Pseudomonas sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
62.3 | - |
Tm of KES24511 in 50 mM Tris chloride, pH 8.0, 200 mM NaCl | Pseudomonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | PLP, the the PLP cofactor is typically covalently bound in the active site to a lysine residue in an internal aldimine conformation | Pseudomonas sp. |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme KES24511 is involved in the amino alkanoate catabolism of the 12-aminododecanoic acid-metabolizing Pseudomonas sp. strain AAC | Pseudomonas sp. |