BRENDA - Enzyme Database
show all sequences of 1.1.99.31

Role of arginine 277 in (S)-mandelate dehydrogenase from Pseudomonas putida in substrate binding and transition state stabilization

Lehoux, I.E.; Mitra, B.; Biochemistry 39, 10055-10065 (2000) View publication on PubMed

Data extracted from this reference:

Engineering
Protein Variants
Commentary
Organism
R277G
kcat for (S)-mandelate is 178fold lower than wild-type value, KM-value for (S)-mandelate is 160fold higher than wild-type value, KM-value for {(S)-3-phenyllactate is 1.6fold lower than wild-type value
Pseudomonas putida
R277H
kcat for (S)-mandelate is 1000fold lower than wild-type value, KM-value for (S)-mandelate is 608fold higher than wild-type value
Pseudomonas putida
R277K
kcat for (S)-mandelate is 3.7fold lower than wild-type value, KM-value for (S)-mandelate is 36.7fold higher than wild-type value
Pseudomonas putida
R277L
kcat for (S)-mandelate is 421fold lower fold than wild-type value, KM-value for (S)-mandelate is 392fold higher than wild-type value
Pseudomonas putida
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.074
-
(S)-3-phenyllactate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
0.12
-
(S)-Mandelate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
4.3
-
(S)-3-phenyllactate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
4.4
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277K
Pseudomonas putida
19.2
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277H
Pseudomonas putida
47
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
73
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277L
Pseudomonas putida
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas putida
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
mutant enzymes R277K, R277G, R277H and R277L
Pseudomonas putida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
(S)-3-phenyllactate + acceptor
-
667567
Pseudomonas putida
?
-
-
-
?
(S)-mandelate + acceptor
-
667567
Pseudomonas putida
2-oxo-2-phenylacetate + reduced acceptor
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.27
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277L
Pseudomonas putida
0.64
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
1.52
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277H
Pseudomonas putida
73
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277K
Pseudomonas putida
201
-
(S)-3-phenyllactate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
270
-
(S)-Mandelate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
270
-
(S)-3-phenyllactate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
Engineering (protein specific)
Protein Variants
Commentary
Organism
R277G
kcat for (S)-mandelate is 178fold lower than wild-type value, KM-value for (S)-mandelate is 160fold higher than wild-type value, KM-value for {(S)-3-phenyllactate is 1.6fold lower than wild-type value
Pseudomonas putida
R277H
kcat for (S)-mandelate is 1000fold lower than wild-type value, KM-value for (S)-mandelate is 608fold higher than wild-type value
Pseudomonas putida
R277K
kcat for (S)-mandelate is 3.7fold lower than wild-type value, KM-value for (S)-mandelate is 36.7fold higher than wild-type value
Pseudomonas putida
R277L
kcat for (S)-mandelate is 421fold lower fold than wild-type value, KM-value for (S)-mandelate is 392fold higher than wild-type value
Pseudomonas putida
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.074
-
(S)-3-phenyllactate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
0.12
-
(S)-Mandelate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
4.3
-
(S)-3-phenyllactate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
4.4
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277K
Pseudomonas putida
19.2
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277H
Pseudomonas putida
47
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
73
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277L
Pseudomonas putida
Purification (Commentary) (protein specific)
Commentary
Organism
mutant enzymes R277K, R277G, R277H and R277L
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
(S)-3-phenyllactate + acceptor
-
667567
Pseudomonas putida
?
-
-
-
?
(S)-mandelate + acceptor
-
667567
Pseudomonas putida
2-oxo-2-phenylacetate + reduced acceptor
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.27
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277L
Pseudomonas putida
0.64
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
1.52
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277H
Pseudomonas putida
73
-
(S)-Mandelate
pH 7.5, 20°C, mutant enzyme R277K
Pseudomonas putida
201
-
(S)-3-phenyllactate
pH 7.5, 20°C, mutant enzyme R277G
Pseudomonas putida
270
-
(S)-Mandelate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
270
-
(S)-3-phenyllactate
pH 7.5, 20°C, wild-type enzyme
Pseudomonas putida
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
-
2
-
-
1
-
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-
1
-
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
-
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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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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
-
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-
-
-
2
-
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1
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-
-
-
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
-
-
-
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-
-
1
-
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-
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-
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
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
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-
-
1
-
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-
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-
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-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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
-
-
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-
-
-
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-
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-
3
-
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-
-
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
Biochem. J.
333
107-115
1998
-
-
1
-
-
-
-
6
2
-
-
-
-
8
-
-
1
-
-
-
-
-
3
-
1
-
-
-
8
-
-
-
-
-
-
-
-
-
1
-
-
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-
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-
6
2
-
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1
-
-
-
-
3
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
667457
Smekal
Substrate analogues as probes ...
Rhodotorula graminis
Biochem. J.
297
647-652
1994
-
-
-
-
-
-
-
11
-
-
-
-
-
3
-
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-
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-
11
-
-
-
-
-
12
-
-
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-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
-
-
-
-
11
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
667455
Smekal
L-mandelate dehydrogenase from ...
Rhodotorula graminis
Biochem. J.
290
103-107
1993
-
-
-
-
-
-
4
3
-
-
-
-
-
7
-
-
-
-
-
-
-
-
3
-
2
-
-
-
3
-
-
-
1
4
-
-
-
-
-
1
-
-
-
-
4
4
3
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
667564
Mitra
A novel structural basis for m ...
Pseudomonas putida
Biochemistry
32
12959-12967
1993
-
-
1
-
1
1
-
6
2
-
-
-
-
6
-
-
1
-
-
-
-
1
2
-
1
-
-
-
4
-
-
-
1
-
-
-
-
-
1
1
-
1
1
-
-
-
6
2
-
-
-
-
-
-
1
-
-
-
1
2
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
669678
Fewson
Comparison of mandelate dehydr ...
Acinetobacter calcoaceticus
J. Gen. Microbiol.
134
967-974
1988
-
-
-
-
-
-
1
-
-
-
-
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-
4
-
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-
-
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-
1
-
2
-
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1
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-
-
1
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1
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1
-
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