Literature summary for 2.6.1.82 extracted from

Wilding, M.; Scott, C.; Newman, J.; Peat, T.S.
Crystal structure of a putrescine aminotransferase from Pseudomonas sp. strain AAC (2017), Acta Crystallogr. Sect. F, 73, 29-35 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant His-tagged enzyme expression in Escherichia coli strain Rosetta 2 (DE3) as soluble protein	Pseudomonas sp.

Crystallization (Commentary)

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.

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

Inhibitors	Comment	Organism	Structure
gabaculine	a transaminase suicide inhibitor	Pseudomonas sp.

Natural Substrates/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Pseudomonas sp.	-	-	-
Pseudomonas sp. AAC	-	-	-

Purification (Commentary)

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Synonyms	Comment	Organism
FG99_07980	-	Pseudomonas sp.
KES24511	-	Pseudomonas sp.
putrescine aminotransferase	-	Pseudomonas sp.

Temperature Stability [°C]

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

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

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.