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Literature summary for 4.1.2.49 extracted from

  • Fesko, K.; Suplatov, D.; Svedas, V.
    Bioinformatic analysis of the fold type I PLP-dependent enzymes reveals determinants of reaction specificity in L-threonine aldolase from Aeromonas jandaei (2018), FEBS open bio, 8, 1013-1028 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.2.49

Protein Variants

Protein Variants Comment Organism
D168N mutation reduces the specific activity, which is partially restored by the addition of excess pyridoxal 5'-phosphate to the reaction mixture. The mutant enzyme is able to catalyse the aldol synthesis of L-beta-phenylserine from benzaldehyde and glycine with yields similar to the wild-type enzyme Aeromonas jandaei
D168S mutation reduces the specific activity, which is partially restored by the addition of excess pyridoxal 5'-phosphate to the reaction mixture. The mutant enzyme is able to catalyse the aldol synthesis of L-beta-phenylserine from benzaldehyde and glycine with yields similar to the wild-type enzyme Aeromonas jandaei
D168T mutation reduces the specific activity, which is partially restored by the addition of excess pyridoxal 5'-phosphate to the reaction mixture. The mutant enzyme is able to catalyse the aldol synthesis of L-beta-phenylserine from benzaldehyde and glycine with yields similar to the wild-type enzyme Aeromonas jandaei
D168V mutation reduces the specific activity, which is partially restored by the addition of excess pyridoxal 5'-phosphate to the reaction mixture. The mutant enzyme is able to catalyse the aldol synthesis of L-beta-phenylserine from benzaldehyde and glycine with yields similar to the wild-type enzyme Aeromonas jandaei
K199R mutation reduces the specific activity of the enzyme by 5000fold, which can be partially restored by adding an excess of pyridoxal 5'-phosphate to the reaction medium Aeromonas jandaei
R171F loss of specific activity by more than 2000fold. pyridoxal 5'-phosphate-glycine complex is not obtained for the mutant. The mutant is not active in the aldol condensation reaction to produce L-beta-phenylserine Aeromonas jandaei
R231A the mutant enzyme racemizes L-alanine to D-alanine, whereas the wild-type enzyme is not active towards L-alanine Aeromonas jandaei
R313H catalytic activity of the mutant in the retro-aldol cleavage of L-allo-threonine is decreased by 200fold mainly due to increase in Km. Mutation hinders the formation of the quinonoid complex to stabilize the carbanion after initial bond cleavage. L-beta-Phenylserine product in the aldol condensation is obtained with the conversion rate similar to the wild-type enzyme Aeromonas jandaei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei
0.5
-
 L-allo-threonine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
0.5
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei
0.8
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
1
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
1.2
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
1.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
1.3
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
2.1
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei
2.3
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
2.6
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168N Aeromonas jandaei
2.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
2.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme K199R Aeromonas jandaei
3.7
-
 L-beta-Phenylserine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
3.7
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei
3.8
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
3.8
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
4.1
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
4.1
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
4.6
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
4.8
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
5.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
5.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
5.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
7
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
7.5
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
11.5
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
14.8
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
17
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei
18
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
28
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
69
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei

Organism

Organism UniProt Comment Textmining
Aeromonas jandaei O07051
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glycine + acetaldehyde
-
 Aeromonas jandaei L-allo-threonine
-
 r
glycine + benzyladehyde
-
 Aeromonas jandaei L-beta-phenylserine
-
 r
L-allo-threonine
-
 Aeromonas jandaei glycine + acetaldehyde
-
 r
L-beta-phenylserine
-
 Aeromonas jandaei glycine + benzyladehyde
-
 r

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
0.02
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
0.05
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168N Aeromonas jandaei
0.05
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
0.08
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme K199R Aeromonas jandaei
0.08
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
0.17
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
0.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
0.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
0.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei
0.23
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
0.3
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
0.3
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
0.32
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
0.42
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
0.47
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei
0.6
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
0.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
0.83
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
1
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
1.9
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
2.3
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
2.3
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
3.9
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
4.3
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
5.1
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
8.67
-
 L-allo-threonine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
13.3
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei
13.6
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei
60.5
-
 L-beta-Phenylserine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
74.4
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei
84
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on Aeromonas jandaei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0007
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
0.0045
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
0.0068
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei
0.01
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
0.011
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171Q Aeromonas jandaei
0.02
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168N Aeromonas jandaei
0.024
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R171F Aeromonas jandaei
0.03
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme K199R Aeromonas jandaei
0.05
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85Y Aeromonas jandaei
0.06
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
0.062
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
0.063
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme H85F Aeromonas jandaei
0.067
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
0.08
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64E Aeromonas jandaei
0.08
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R313H Aeromonas jandaei
0.09
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
0.2
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
0.3
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
0.3
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme N64I Aeromonas jandaei
0.35
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme D168S Aeromonas jandaei
0.4
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme D168V Aeromonas jandaei
0.6
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme R231A Aeromonas jandaei
0.9
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
0.9
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme S198H Aeromonas jandaei
1.6
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
1.7
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme E138A Aeromonas jandaei
16
-
 L-beta-Phenylserine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
17
-
 L-allo-threonine pH 8.0, 25°C, wild-type enzyme Aeromonas jandaei
22
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei
26
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme C196T Aeromonas jandaei
35
-
 L-beta-Phenylserine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei
61
-
 L-allo-threonine pH 8.0, 25°C, mutant enzyme G200S Aeromonas jandaei