Literature summary for 2.6.1.62 extracted from

Bezsudnova, E.Y.; Stekhanova, T.N.; Popinako, A.V.; Rakitina, T.V.; Nikolaeva, A.Y.; Boyko, K.M.; Popov, V.O.
Diaminopelargonic acid transaminase from Psychrobacter cryohalolentis is active towards (S)-(-)-1-phenylethylamine, aldehydes and alpha-diketones (2018), Appl. Microbiol. Biotechnol., 102, 9621-9633 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene Pcryo-0361, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21Star	Psychrobacter cryohalolentis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, sitting drop vapor diffusion method, mixing of 11 mg/ml protein in 50 mM Tris, 100 mM NaCl, and 0.02 mM PLP, pH 7.9, with precipitant solution containing 0.1 M sodium citrate, pH 6.1, 0.2 M potassium sodium tartrate, and 1.8 M ammonium sulfate. A glass capillary containing 0.008 ml of protein solution is mounted in the gel tube with 1% agarose presoaked in 1 ml of the precipitant solution, method optimization, 1 month at 20°C. X-ray diffraction structure determination and analysis at 1.6 A resolution	Psychrobacter cryohalolentis

Crystallization (Comment)

Organism

purified recombinant enzyme, sitting drop vapor diffusion method, mixing of 11 mg/ml protein in 50 mM Tris, 100 mM NaCl, and 0.02 mM PLP, pH 7.9, with precipitant solution containing 0.1 M sodium citrate, pH 6.1, 0.2 M potassium sodium tartrate, and 1.8 M ammonium sulfate. A glass capillary containing 0.008 ml of protein solution is mounted in the gel tube with 1% agarose presoaked in 1 ml of the precipitant solution, method optimization, 1 month at 20°C. X-ray diffraction structure determination and analysis at 1.6 A resolution

Psychrobacter cryohalolentis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	kinetic analysis of the half-reaction with different substrates	Psychrobacter cryohalolentis
0.22	-	2,3-Butanedione	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis
1.4	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with isobutyraldehyde	Psychrobacter cryohalolentis
3.87	-	Isobutyraldehyde	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis
8.2	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with 2,3-butanedione	Psychrobacter cryohalolentis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	Psychrobacter cryohalolentis	-	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	Psychrobacter cryohalolentis DSM 17306	-	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	Psychrobacter cryohalolentis K5	-	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	Psychrobacter cryohalolentis VKM B-2378	-	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	Psychrobacter cryohalolentis ATCC BAA-1226	-	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?

Organism

Organism	UniProt	Comment	Textmining
Psychrobacter cryohalolentis	Q1QDV8	-	-
Psychrobacter cryohalolentis ATCC BAA-1226	Q1QDV8	-	-
Psychrobacter cryohalolentis DSM 17306	Q1QDV8	-	-
Psychrobacter cryohalolentis K5	Q1QDV8	-	-
Psychrobacter cryohalolentis VKM B-2378	Q1QDV8	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2,3-butanedione + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis	?	-	?
2,3-butanedione + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis DSM 17306	?	-	?
2,3-butanedione + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis K5	?	-	?
2,3-butanedione + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis VKM B-2378	?	-	?
2,3-butanedione + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis ATCC BAA-1226	?	-	?
2,3-dihydroxypropanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
2,3-dihydroxypropanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis DSM 17306	?	-	?
2,3-dihydroxypropanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis K5	?	-	?
2,3-dihydroxypropanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis VKM B-2378	?	-	?
2,3-dihydroxypropanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis ATCC BAA-1226	?	-	?
2,3-hexanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
2,3-hexanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis DSM 17306	?	-	?
2,3-hexanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis K5	?	-	?
2,3-hexanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis VKM B-2378	?	-	?
2,3-hexanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis ATCC BAA-1226	?	-	?
2,3-pentanedione + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
2-oxo-2-phenylacetaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
benzaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	propiophenone + ?	-	?
glyoxylate + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
hexanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
isobutyraldehyde + (S)-(-)-1-phenylethylamine	molecular docking study	Psychrobacter cryohalolentis	acetophenone + ?	-	?
isobutyraldehyde + (S)-(-)-1-phenylpropylamine	-	Psychrobacter cryohalolentis	acetophenone + ?	-	?
additional information	7,8-diaminopelargonic acid transaminase (synthase), previously considered to be highly specific, is able to convert (S)-(-)-1-phenylethylamine and a number of aldehydes and diketones. Only a weak activity towards pyruvate is detected. The enzyme shows (S)-amine transaminase activity. The Pcryo361 affinity towards (S)-(-)-1-phenylethylamine arises from the recognition of the hydrophobic parts of the specific substrates, S-adenosyl-L-methionine and 7-keto-8-aminopelargonic acid, and from the flexibility of the active site. Analysis of the half-transamination reaction measuring a decrease in aldimine concentration in the presence of different concentrations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, L-lysine, and (S)-(-)-1,2-diaminopropane, substrate specificity of enzyme Pcryo361 in the half and the overall reactions, overview. No activity towards 2,4-pentanedione, 2-oxobutyrate, or acetone as amine acceptor substrates, poor activity with pyruvate, no activity with (R)-(-)-1-phenylethylamine as amine donor substrate	Psychrobacter cryohalolentis	?	-	-
additional information	7,8-diaminopelargonic acid transaminase (synthase), previously considered to be highly specific, is able to convert (S)-(-)-1-phenylethylamine and a number of aldehydes and diketones. Only a weak activity towards pyruvate is detected. The enzyme shows (S)-amine transaminase activity. The Pcryo361 affinity towards (S)-(-)-1-phenylethylamine arises from the recognition of the hydrophobic parts of the specific substrates, S-adenosyl-L-methionine and 7-keto-8-aminopelargonic acid, and from the flexibility of the active site. Analysis of the half-transamination reaction measuring a decrease in aldimine concentration in the presence of different concentrations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, L-lysine, and (S)-(-)-1,2-diaminopropane, substrate specificity of enzyme Pcryo361 in the half and the overall reactions, overview. No activity towards 2,4-pentanedione, 2-oxobutyrate, or acetone as amine acceptor substrates, poor activity with pyruvate, no activity with (R)-(-)-1-phenylethylamine as amine donor substrate	Psychrobacter cryohalolentis DSM 17306	?	-	-
additional information	7,8-diaminopelargonic acid transaminase (synthase), previously considered to be highly specific, is able to convert (S)-(-)-1-phenylethylamine and a number of aldehydes and diketones. Only a weak activity towards pyruvate is detected. The enzyme shows (S)-amine transaminase activity. The Pcryo361 affinity towards (S)-(-)-1-phenylethylamine arises from the recognition of the hydrophobic parts of the specific substrates, S-adenosyl-L-methionine and 7-keto-8-aminopelargonic acid, and from the flexibility of the active site. Analysis of the half-transamination reaction measuring a decrease in aldimine concentration in the presence of different concentrations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, L-lysine, and (S)-(-)-1,2-diaminopropane, substrate specificity of enzyme Pcryo361 in the half and the overall reactions, overview. No activity towards 2,4-pentanedione, 2-oxobutyrate, or acetone as amine acceptor substrates, poor activity with pyruvate, no activity with (R)-(-)-1-phenylethylamine as amine donor substrate	Psychrobacter cryohalolentis K5	?	-	-
additional information	7,8-diaminopelargonic acid transaminase (synthase), previously considered to be highly specific, is able to convert (S)-(-)-1-phenylethylamine and a number of aldehydes and diketones. Only a weak activity towards pyruvate is detected. The enzyme shows (S)-amine transaminase activity. The Pcryo361 affinity towards (S)-(-)-1-phenylethylamine arises from the recognition of the hydrophobic parts of the specific substrates, S-adenosyl-L-methionine and 7-keto-8-aminopelargonic acid, and from the flexibility of the active site. Analysis of the half-transamination reaction measuring a decrease in aldimine concentration in the presence of different concentrations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, L-lysine, and (S)-(-)-1,2-diaminopropane, substrate specificity of enzyme Pcryo361 in the half and the overall reactions, overview. No activity towards 2,4-pentanedione, 2-oxobutyrate, or acetone as amine acceptor substrates, poor activity with pyruvate, no activity with (R)-(-)-1-phenylethylamine as amine donor substrate	Psychrobacter cryohalolentis VKM B-2378	?	-	-
additional information	7,8-diaminopelargonic acid transaminase (synthase), previously considered to be highly specific, is able to convert (S)-(-)-1-phenylethylamine and a number of aldehydes and diketones. Only a weak activity towards pyruvate is detected. The enzyme shows (S)-amine transaminase activity. The Pcryo361 affinity towards (S)-(-)-1-phenylethylamine arises from the recognition of the hydrophobic parts of the specific substrates, S-adenosyl-L-methionine and 7-keto-8-aminopelargonic acid, and from the flexibility of the active site. Analysis of the half-transamination reaction measuring a decrease in aldimine concentration in the presence of different concentrations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, L-lysine, and (S)-(-)-1,2-diaminopropane, substrate specificity of enzyme Pcryo361 in the half and the overall reactions, overview. No activity towards 2,4-pentanedione, 2-oxobutyrate, or acetone as amine acceptor substrates, poor activity with pyruvate, no activity with (R)-(-)-1-phenylethylamine as amine donor substrate	Psychrobacter cryohalolentis ATCC BAA-1226	?	-	-
propanaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
pyruvaldehyde + (S)-(-)-1-phenylethylamine	-	Psychrobacter cryohalolentis	?	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	-	Psychrobacter cryohalolentis	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	-	Psychrobacter cryohalolentis DSM 17306	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	-	Psychrobacter cryohalolentis K5	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	-	Psychrobacter cryohalolentis VKM B-2378	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?
S-adenosyl-L-methionine + 8-amino-7-oxononanoate	-	Psychrobacter cryohalolentis ATCC BAA-1226	S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate	-	?

Synonyms

Synonyms	Comment	Organism
7,8-diaminopelargonic acid synthase	-	Psychrobacter cryohalolentis
7,8-diaminopelargonic acid transaminase	-	Psychrobacter cryohalolentis
diaminopelargonic acid transaminase	-	Psychrobacter cryohalolentis
More	cf. EC 2.6.1, (S)-specific amine transaminase ((S)-ATA)	Psychrobacter cryohalolentis
Pcryo_0361	-	Psychrobacter cryohalolentis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
35	42	-	Psychrobacter cryohalolentis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.026	-	2,3-Butanedione	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis
0.045	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with 2,3-butanedione	Psychrobacter cryohalolentis
0.16	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with isobutyraldehyde	Psychrobacter cryohalolentis
0.25	-	Isobutyraldehyde	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
9	11	-	Psychrobacter cryohalolentis

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	PLP	Psychrobacter cryohalolentis

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to class III of the superfamily of aminotransferases (transaminase, TA)	Psychrobacter cryohalolentis
additional information	the conversion of amines is a promiscuous activity of many transaminases of class III and is independent from their natural function. Molecular docking calculations of S-adenosyl-L-methionine, (S)-(-)-1-phenylethylamine, and its quinonoid intermediates in the active site of the enzyme	Psychrobacter cryohalolentis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.0055	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with 2,3-butanedione	Psychrobacter cryohalolentis
0.065	-	Isobutyraldehyde	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis
0.11	-	(S)-(-)-1-phenylethylamine	pH 8.0, 35°C, with isobutyraldehyde	Psychrobacter cryohalolentis
0.12	-	2,3-Butanedione	pH 8.0, 35°C, with (S)-(-)-1-phenylethylamine	Psychrobacter cryohalolentis