BRENDA - Enzyme Database
show all sequences of 1.1.99.31

(S)-Mandelate dehydrogenase from Pseudomonas putida: mechanistic studies with alternate substrates and pH and kinetic isotope effects

Lehoux, I.E.; Mitra, B.; Biochemistry 38, 5836-5848 (1999)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed in Escherichia coli
Pseudomonas putida
Inhibitors
Inhibitors
Commentary
Organism
Structure
(R,S)-2-hydroxy-3-butynoic acid
irreversible inactivation
Pseudomonas putida
(S)-1-phenyl-1,2-ethanediol
competitive inhibitor
Pseudomonas putida
(S)-2-methoxy-2-phenylacetate
competitive inhibitor
Pseudomonas putida
(S)-3-phenyllactate
competitive inhibitor
Pseudomonas putida
2-phenylacetate
competitive inhibitor
Pseudomonas putida
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.33
-
(R,S)-mandelate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.4
-
(R,S)-indoleglycolate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.75
-
(R,S)-2-hydroxyoctanoate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.9
-
(R,S)-3-indolelactate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
2
-
DL-3-phenyllactate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
4.3
-
(R,S)-2-hydroxyisocaproate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
4.9
-
(R,S)-2-hydroxyhexanoate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
9.5
-
(R,S)-2-hydroxyisovalerate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
15.3
-
(R,S)-2-hydroxyvalerate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
22
-
(R,S)-2-hydroxy-3-butynoic acid
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
32
-
(R,S)-2-hydroxybutyrate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas putida
P20932
strain ATCC 12633
-
Purification (Commentary)
Purification (Commentary)
Organism
His6-tagged enzyme, Ni-resin column chromatography and Sephadex G-25 gel filtration
Pseudomonas putida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
(R,S)-2-hydroxy-3-butenoic acid + acceptor
very low binding affinity for MDH
678122
Pseudomonas putida
2-oxo-3-butenoic acid + reduced acceptor
-
-
-
?
(R,S)-2-hydroxybutyrate + acceptor
slow substrate
678122
Pseudomonas putida
2-oxobutyrate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxybutyric acid + acceptor
very low binding affinity for MDH
678122
Pseudomonas putida
2-oxo-3-butynoic acid + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyhexanoate + acceptor
-
678122
Pseudomonas putida
2-oxohexanoate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyisocaproate + acceptor
-
678122
Pseudomonas putida
2-oxoisocaproate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyisovalerate + acceptor
-
678122
Pseudomonas putida
2-oxo-isovalerate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyoctanoate + acceptor
slow substrate
678122
Pseudomonas putida
2-oxooctanoate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyvalerate + acceptor
-
678122
Pseudomonas putida
2-oxovalerate + reduced acceptor
-
-
-
?
(R,S)-3-indolelactate + acceptor
-
678122
Pseudomonas putida
3-(1H-indol-2-yl)-2-oxopropanoate + reduced acceptor
-
-
-
?
(R,S)-3-phenyllactate + 2,6-dichlorophenolindophenol
slow substrate
678122
Pseudomonas putida
phenylpyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R,S)-indoleglycolate + acceptor
-
678122
Pseudomonas putida
indol-2-yl(oxo)acetate + reduced acceptor
-
-
-
?
(R,S)-mandelate + 2,6-dichlorophenolindophenol
-
678122
Pseudomonas putida
2-oxo-2-phenylacetate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R,S)-mandelate + ferricyanide
-
678122
Pseudomonas putida
2-oxo-2-phenylacetate + ferrocyanide
-
-
-
?
2-hydroxy-3-butynoate + acceptor
-
678122
Pseudomonas putida
2-oxo-3-butynoate + reduced acceptor
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
MDH
-
Pseudomonas putida
Cofactor
Cofactor
Commentary
Organism
Structure
FMN
-
Pseudomonas putida
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
36
-
(R,S)-2-hydroxy-3-butynoic acid
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli
Pseudomonas putida
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FMN
-
Pseudomonas putida
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
(R,S)-2-hydroxy-3-butynoic acid
irreversible inactivation
Pseudomonas putida
(S)-1-phenyl-1,2-ethanediol
competitive inhibitor
Pseudomonas putida
(S)-2-methoxy-2-phenylacetate
competitive inhibitor
Pseudomonas putida
(S)-3-phenyllactate
competitive inhibitor
Pseudomonas putida
2-phenylacetate
competitive inhibitor
Pseudomonas putida
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
36
-
(R,S)-2-hydroxy-3-butynoic acid
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.33
-
(R,S)-mandelate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.4
-
(R,S)-indoleglycolate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.75
-
(R,S)-2-hydroxyoctanoate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
0.9
-
(R,S)-3-indolelactate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
2
-
DL-3-phenyllactate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
4.3
-
(R,S)-2-hydroxyisocaproate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
4.9
-
(R,S)-2-hydroxyhexanoate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
9.5
-
(R,S)-2-hydroxyisovalerate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
15.3
-
(R,S)-2-hydroxyvalerate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
22
-
(R,S)-2-hydroxy-3-butynoic acid
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
32
-
(R,S)-2-hydroxybutyrate
in 0.1 M potassium phosphate buffer, pH 7.5 at 20°C
Pseudomonas putida
Purification (Commentary) (protein specific)
Commentary
Organism
His6-tagged enzyme, Ni-resin column chromatography and Sephadex G-25 gel filtration
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
(R,S)-2-hydroxy-3-butenoic acid + acceptor
very low binding affinity for MDH
678122
Pseudomonas putida
2-oxo-3-butenoic acid + reduced acceptor
-
-
-
?
(R,S)-2-hydroxybutyrate + acceptor
slow substrate
678122
Pseudomonas putida
2-oxobutyrate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxybutyric acid + acceptor
very low binding affinity for MDH
678122
Pseudomonas putida
2-oxo-3-butynoic acid + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyhexanoate + acceptor
-
678122
Pseudomonas putida
2-oxohexanoate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyisocaproate + acceptor
-
678122
Pseudomonas putida
2-oxoisocaproate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyisovalerate + acceptor
-
678122
Pseudomonas putida
2-oxo-isovalerate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyoctanoate + acceptor
slow substrate
678122
Pseudomonas putida
2-oxooctanoate + reduced acceptor
-
-
-
?
(R,S)-2-hydroxyvalerate + acceptor
-
678122
Pseudomonas putida
2-oxovalerate + reduced acceptor
-
-
-
?
(R,S)-3-indolelactate + acceptor
-
678122
Pseudomonas putida
3-(1H-indol-2-yl)-2-oxopropanoate + reduced acceptor
-
-
-
?
(R,S)-3-phenyllactate + 2,6-dichlorophenolindophenol
slow substrate
678122
Pseudomonas putida
phenylpyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R,S)-indoleglycolate + acceptor
-
678122
Pseudomonas putida
indol-2-yl(oxo)acetate + reduced acceptor
-
-
-
?
(R,S)-mandelate + 2,6-dichlorophenolindophenol
-
678122
Pseudomonas putida
2-oxo-2-phenylacetate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R,S)-mandelate + ferricyanide
-
678122
Pseudomonas putida
2-oxo-2-phenylacetate + ferrocyanide
-
-
-
?
2-hydroxy-3-butynoate + acceptor
-
678122
Pseudomonas putida
2-oxo-3-butynoate + reduced acceptor
-
-
-
?
Other publictions for EC 1.1.99.31
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
740174
Chen
Stereoselective biotransformat ...
Escherichia coli
Bioresour. Bioproc.
4
2-2
2017
-
-
-
-
-
-
-
1
-
-
-
1
-
1
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1
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1
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1
1
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1
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1
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1
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1
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1
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1
-
-
-
1
-
-
-
-
-
-
-
-
-
740119
Mazurenko
Immobilization of membrane-bou ...
Pseudomonas putida, Pseudomonas putida DSM 291
Bioelectrochemistry
104
65-70
2015
-
-
-
-
-
-
-
-
-
-
-
2
-
4
-
-
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-
-
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2
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1
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-
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2
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2
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-
-
-
-
-
-
-
-
725744
Wang
-
A bi-enzymatic system for effi ...
Pseudomonas aeruginosa
J. Mol. Catal. B
94
47-50
2013
-
-
1
-
-
-
-
2
-
-
-
1
-
1
-
-
1
-
-
-
2
-
1
-
1
1
1
1
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
2
-
-
-
1
-
-
-
1
-
-
2
-
1
-
1
1
1
-
1
-
-
-
-
-
-
-
-
-
695329
Sukumar
Structures of the G81A mutant ...
Pseudomonas putida
Acta Crystallogr. Sect. D
65
543-552
2009
-
-
1
1
2
-
-
12
-
-
-
-
-
4
-
-
1
-
-
-
-
-
4
-
1
-
-
-
12
-
-
-
1
-
-
-
-
-
1
1
1
2
-
-
-
-
12
-
-
-
-
-
-
-
1
-
-
-
-
4
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
667577
Dewanti
Role of glycine 81 in (S)-mand ...
Pseudomonas putida
Biochemistry
43
10692-10700
2004
-
-
-
-
5
-
1
36
-
-
-
-
-
4
-
-
1
-
-
-
-
-
13
-
-
-
-
-
36
-
-
-
1
-
-
-
-
-
-
1
-
5
-
-
1
-
36
-
-
-
-
-
-
-
1
-
-
-
-
13
-
-
-
-
36
-
-
-
-
-
-
-
-
-
-
669259
Sukumar
High resolution structures of ...
Pseudomonas putida
J. Biol. Chem.
279
3749-3757
2004
-
-
-
1
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
678128
Dewanti
Esters of mandelic acid as sub ...
Pseudomonas putida
Biochemistry
43
1883-1890
2004
-
-
-
-
2
-
4
9
-
-
-
-
-
5
-
-
1
-
-
-
-
-
4
-
1
-
-
-
9
-
-
-
1
2
-
-
-
-
-
1
-
2
-
-
4
2
9
-
-
-
-
-
-
-
1
-
-
-
-
4
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
667575
Dewanti
A transient intermediate in th ...
Pseudomonas putida
Biochemistry
42
12893-12901
2003
-
-
-
-
-
-
-
2
-
-
-
-
-
5
-
-
1
-
-
-
-
-
1
-
-
-
-
-
2
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
667572
Xu
Arginine 165/arginine 277 pair ...
Pseudomonas putida
Biochemistry
41
12313-12319
2002
-
-
-
-
10
-
3
9
-
-
-
-
-
5
-
-
1
-
-
-
-
1
1
-
-
-
-
-
9
-
-
-
1
13
-
-
-
-
-
1
-
10
-
-
3
13
9
-
-
-
-
-
-
-
1
-
-
-
1
1
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
667570
Sukumar
Structure of an active soluble ...
Pseudomonas putida
Biochemistry
40
9870-9878
2001
-
-
-
1
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667567
Lehoux
Role of arginine 277 in (S)-ma ...
Pseudomonas putida
Biochemistry
39
10055-10065
2000
-
-
-
-
4
-
-
7
-
-
-
-
-
4
-
-
1
-
-
-
-
-
2
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
7
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
667565
Xu
A highly active, soluble mutan ...
Pseudomonas putida
Biochemistry
38
12367-12376
1999
-
-
-
-
1
-
-
14
1
-
-
-
-
6
-
-
-
-
-
-
-
-
14
-
1
-
-
-
15
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
14
1
-
-
-
-
-
-
-
-
-
-
-
14
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
667566
Lehoux
(S)-Mandelate dehydrogenase fr ...
Pseudomonas putida
Biochemistry
38
9948-9955
1999
-
-
-
-
3
-
-
1
-
-
-
-
-
5
-
-
1
-
-
-
-
-
2
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
678122
Lehoux
(S)-Mandelate dehydrogenase fr ...
Pseudomonas putida
Biochemistry
38
5836-5848
1999
-
-
1
-
-
-
5
11
-
-
-
-
-
4
-
-
1
-
-
-
-
-
14
-
1
-
-
-
-
-
-
-
1
1
-
-
-
-
1
1
-
-
-
-
5
1
11
-
-
-
-
-
-
-
1
-
-
-
-
14
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667458
Illias
L-Mandelate dehydrogenase from ...
Rhodotorula graminis
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