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Literature summary for 1.1.99.31 extracted from

  • Dewanti, A.R.; Xu, Y.; Mitra, B.
    Role of glycine 81 in (S)-mandelate dehydrogenase from Pseudomonas putida in substrate specificity and oxidase activity (2004), Biochemistry, 43, 10692-10700.
    View publication on PubMed
Search for more literature for 1.1.99.31

Protein Variants

Protein Variants Comment Organism
G81A higher specificity for small substrates, compared to that of wild-type enzyme, the affinity for (S)-mandelate is relatively unchanged. The rate of the first half-reaction is slower than the wild-type rate. 2fold increase in oxidative half-reaction. Affinity for oxygen increases 10-15fold Pseudomonas putida
G81D extreme low activity, reduction in activity is due to the decrease in electrophilicity of the FMN Pseudomonas putida
G81S the rate of the first half-reaction is slower than the wild-type rate. The rate of the first half-reaction is slower than the wild-type rate. Affinity for O2 increases 10-15fold Pseudomonas putida
G81V extreme low activity, reduction in activity is due to the decrease in electrophilicity of the FMN Pseudomonas putida
additional information MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach. G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
sulfite
-
 Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
 O2 pH 7.5, 20°C, mutant enzyme G81V Pseudomonas putida
0.04
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme, G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH Pseudomonas putida
0.09
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach Pseudomonas putida
0.13
-
 (S)-Mandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.14
-
 2-Hydroxyoctanoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.15
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.17
-
 3-indolelactate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.17
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81V Pseudomonas putida
0.22
-
 O2 pH 7.5, 20°C, mutant enzyme G81D Pseudomonas putida
0.24
-
 Mandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.24
-
 Mandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.27
-
 4-chloromandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.37
-
 3-Phenyllactate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.4
-
 indoleglycolate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.43
-
 4-chloromandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.49
-
 O2 pH 7.5, 20°C, mutant enzyme, G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH Pseudomonas putida
0.63
-
 indoleglycolate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.68
-
 O2 pH 7.5, 20°C, mutant enzyme MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with 205 residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach Pseudomonas putida
0.8
-
 2-Hydroxyoctanoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.9
-
 3-indolelactate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
1.1
-
 O2 pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
1.4
-
 2-hydroxyhexanoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
1.4
-
 O2 pH 7.5, 20°C, mutant enzyme G81S Pseudomonas putida
1.6
-
 2-hydroxyisocaproate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
2
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81D Pseudomonas putida
2
-
 3-Phenyllactate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
2.3
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81S Pseudomonas putida
3.2
-
 O2 pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
4.3
-
 2-Hydroxy-3-butynoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
4.3
-
 2-hydroxyisocaproate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
4.4
-
 2-Hydroxybutyrate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
4.9
-
 2-hydroxyhexanoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
6.4
-
 2-hydroxyvalerate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
15.3
-
 2-hydroxyvalerate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
22
-
 2-Hydroxy-3-butynoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
32
-
 2-Hydroxybutyrate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzymes Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-2-hydroxy-2-phenylacetate + O2 in absence of any other electron acceptor, oxygen is used by the wild-type enzyme for reoxidation of the reduced flavin, at a rather slow rate. Ping-pong kinetics Pseudomonas putida 2-oxo-2-phenylacetate + H2O2
-
 ?
(S)-mandelate + acceptor phenazine methosulfate and 2,6-dichloroindophenol as acceptor. In absence of any other electron acceptor, oxygen is used by the wild-type enzyme for reoxidation of the reduced flavin, at a rather slow rate Pseudomonas putida 2-oxo-2-phenylacetate + reduced acceptor
-
 ?
2-hydroxy-3-butynoate + acceptor
-
 Pseudomonas putida 2-oxo-3-butynoate + reduced acceptor
-
 ?
2-hydroxybutyrate + acceptor
-
 Pseudomonas putida 2-oxobutyrate + reduced acceptor
-
 ?
2-hydroxyhexanoate + acceptor
-
 Pseudomonas putida 2-oxohexanoate + reduced acceptor
-
 ?
2-hydroxyisocaproate + acceptor
-
 Pseudomonas putida 2-oxoisocaproate + reduced acceptor
-
 ?
2-hydroxyoctanoate + acceptor
-
 Pseudomonas putida 2-oxooctanoate + reduced acceptor
-
 ?
2-hydroxyvalerate + acceptor
-
 Pseudomonas putida 2-oxovalerate + reduced acceptor
-
 ?
3-indolelactate + acceptor
-
 Pseudomonas putida 3-(1H-indol-2-yl)-2-oxopropanoate + reduced acceptor
-
 ?
3-phenyllactate + acceptor
-
 Pseudomonas putida 2-oxo-3-phenylpropanoate + reduced acceptor
-
 ?
4-chloromandelate + acceptor
-
 Pseudomonas putida 2-oxo-2-(4-chlorophenyl)acetate + reduced acceptor
-
 ?
indoleglycolate + acceptor
-
 Pseudomonas putida indol-2-yl(oxo)acetate + reduced acceptor
-
 ?
mandelate + acceptor
-
 Pseudomonas putida 2-oxo-2-phenylacetate + reduced acceptor
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.03
-
 O2 pH 7.5, 20°C, mutant enzyme MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with 205 residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach Pseudomonas putida
0.05
-
 O2 pH 7.5, 20°C, mutant enzyme G81D Pseudomonas putida
0.05
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81V Pseudomonas putida
0.07
-
 O2 pH 7.5, 20°C, mutant enzyme G81V Pseudomonas putida
0.1
-
 2-Hydroxybutyrate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.11
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81D Pseudomonas putida
0.15
-
 3-Phenyllactate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.19
-
 2-hydroxyhexanoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.22
-
 O2 pH 7.5, 20°C, mutant enzyme, G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH Pseudomonas putida
0.26
-
 2-Hydroxyoctanoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.27
-
 3-indolelactate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.27
-
 2-hydroxyvalerate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.34
-
 2-hydroxyhexanoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.36
-
 2-hydroxyvalerate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.37
-
 2-Hydroxybutyrate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.48
-
 2-hydroxyisocaproate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
0.5
-
 2-Hydroxyoctanoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.52
-
 3-Phenyllactate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
0.61
-
 O2 pH 7.5, 20°C, mutant enzyme G81S Pseudomonas putida
0.8
-
 2-hydroxyisocaproate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
1
-
 3-indolelactate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
1.2
-
 indoleglycolate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
1.2
-
 O2 pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
2.3
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme, G81A/MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach and a G81A mutation in MDH Pseudomonas putida
2.8
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81S Pseudomonas putida
3.4
-
 O2 pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
3.9
-
 2-Hydroxy-3-butynoate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
9.3
-
 4-chloromandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
14.8
-
 2-Hydroxy-3-butynoate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
15.2
-
 Mandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
19.2
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme G81A Pseudomonas putida
122
-
 indoleglycolate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
205
-
 (S)-Mandelate pH 7.5, 20°C, mutant enzyme MDH-GOX2, a chimeric mutant of (S)-mandelate dehydrogenase with 205 residues 177-215 replaced by residues 176-195 of glycolate oxidase from spinach Pseudomonas putida
334
-
 4-chloromandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
350
-
 Mandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida
360
-
 (S)-Mandelate pH 7.5, 20°C, wild-type enzyme Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
FMN dependent on. In the reductive half-reaction (S)-mandelate is oxidized to 2-oxo-2-phenylacetate, resulting in the reduction of FMN Pseudomonas putida