BRENDA - Enzyme Database
show all sequences of 1.20.1.1

Chemical rescue and inhibition studies to determine the role of Arg301 in phosphite dehydrogenase

Hung, J.E.; Fogle, E.J.; Garg, N.; Chekan, J.R.; Nair, S.K.; van der Donk, W.A.; PLoS ONE 9, e87134 (2014)

Data extracted from this reference:

Crystallization (Commentary)
Crystallization
Organism
mutant enzymes R301K and R301A, hanging drop vapor diffusion method, using 0.1 M NaCl, 0.1 M sodium cacodylate pH 5.8, 16% PEG 6000 (w/v) (for mutant R301K) or 0.1 M ammonium acetate, 0.1 M Bis Tris pH 5.5, 17% PEG 10000 (w/v) (for mutant R301A)
Pseudomonas stutzeri
Engineering
Amino acid exchange
Commentary
Organism
R301A
the mutant shows about 100fold loss in kcat and a 500fold increased Km value for phosphonate. The Ki value of sulfite with the mutant is 400fold increased compared to the wild type enzyme
Pseudomonas stutzeri
R301K
the mutant exhibits a slightly higher kcat than the wild type enzyme and a 20fold increased Km value for phosphonate
Pseudomonas stutzeri
Inhibitors
Inhibitors
Commentary
Organism
Structure
sulfite
competitive inhibitor
Pseudomonas stutzeri
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.028
-
phosphonate
wild type enzyme, at pH 7.3 and 25°C
Pseudomonas stutzeri
1.1
-
phosphonate
mutant enzyme R301K, at pH 7.3 and 25°C
Pseudomonas stutzeri
20
-
phosphonate
mutant enzyme R301A, at pH 7.3 and 25°C
Pseudomonas stutzeri
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
phosphonate + NAD+ + H2O
Pseudomonas stutzeri
-
phosphate + NADH + H+
-
-
?
phosphonate + NAD+ + H2O
Pseudomonas stutzeri WM88
-
phosphate + NADH + H+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas stutzeri
O69054
-
-
Pseudomonas stutzeri WM88
O69054
-
-
Purification (Commentary)
Commentary
Organism
Ni2+-affinity column chromatography and Superdex 200 gel filtration
Pseudomonas stutzeri
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
phosphonate + NAD+ + H2O
-
746256
Pseudomonas stutzeri
phosphate + NADH + H+
-
-
-
?
phosphonate + NAD+ + H2O
-
746256
Pseudomonas stutzeri WM88
phosphate + NADH + H+
-
-
-
?
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Pseudomonas stutzeri
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00031
-
sulfite
wild type enzyme, at pH 7.3 and 25°C
Pseudomonas stutzeri
0.078
-
sulfite
mutant enzyme R301K, at pH 7.3 and 25°C
Pseudomonas stutzeri
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Pseudomonas stutzeri
Crystallization (Commentary) (protein specific)
Crystallization
Organism
mutant enzymes R301K and R301A, hanging drop vapor diffusion method, using 0.1 M NaCl, 0.1 M sodium cacodylate pH 5.8, 16% PEG 6000 (w/v) (for mutant R301K) or 0.1 M ammonium acetate, 0.1 M Bis Tris pH 5.5, 17% PEG 10000 (w/v) (for mutant R301A)
Pseudomonas stutzeri
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
R301A
the mutant shows about 100fold loss in kcat and a 500fold increased Km value for phosphonate. The Ki value of sulfite with the mutant is 400fold increased compared to the wild type enzyme
Pseudomonas stutzeri
R301K
the mutant exhibits a slightly higher kcat than the wild type enzyme and a 20fold increased Km value for phosphonate
Pseudomonas stutzeri
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
sulfite
competitive inhibitor
Pseudomonas stutzeri
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00031
-
sulfite
wild type enzyme, at pH 7.3 and 25°C
Pseudomonas stutzeri
0.078
-
sulfite
mutant enzyme R301K, at pH 7.3 and 25°C
Pseudomonas stutzeri
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.028
-
phosphonate
wild type enzyme, at pH 7.3 and 25°C
Pseudomonas stutzeri
1.1
-
phosphonate
mutant enzyme R301K, at pH 7.3 and 25°C
Pseudomonas stutzeri
20
-
phosphonate
mutant enzyme R301A, at pH 7.3 and 25°C
Pseudomonas stutzeri
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
phosphonate + NAD+ + H2O
Pseudomonas stutzeri
-
phosphate + NADH + H+
-
-
?
phosphonate + NAD+ + H2O
Pseudomonas stutzeri WM88
-
phosphate + NADH + H+
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
Ni2+-affinity column chromatography and Superdex 200 gel filtration
Pseudomonas stutzeri
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
phosphonate + NAD+ + H2O
-
746256
Pseudomonas stutzeri
phosphate + NADH + H+
-
-
-
?
phosphonate + NAD+ + H2O
-
746256
Pseudomonas stutzeri WM88
phosphate + NADH + H+
-
-
-
?
Other publictions for EC 1.20.1.1
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
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)
744126
Beyer
P450BM3 fused to phosphite de ...
Pseudomonas stutzeri, Pseudomonas stutzeri WM88
Appl. Microbiol. Biotechnol.
101
2319-2331
2017
-
-
1
-
-
-
-
1
-
-
-
2
-
5
-
-
1
-
-
-
-
-
2
-
-
-
-
1
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
1
-
-
-
2
-
-
-
1
-
-
-
-
2
-
-
-
-
1
-
-
-
-
-
-
-
-
1
1
744691
Ranaghan
-
A catalytic role for methioni ...
Pseudomonas stutzeri, Pseudomonas stutzeri WM88
Chem. Sci.
5
2191-2199
2014
-
-
-
-
3
-
1
6
-
-
-
2
-
3
-
-
-
-
-
-
-
-
2
-
-
-
-
3
-
-
-
1
-
-
-
-
-
-
1
-
3
-
-
1
-
6
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
3
-
-
-
-
-
-
-
-
3
3
745418
Kanda
Application of a phosphite de ...
Ralstonia sp.
J. Biotechnol.
182-183
68-73
2014
-
-
1
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
746256
Hung
Chemical rescue and inhibitio ...
Pseudomonas stutzeri, Pseudomonas stutzeri WM88
PLoS ONE
9
e87134
2014
-
-
-
1
2
-
1
3
-
-
-
2
-
4
-
-
1
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
2
-
-
-
-
-
1
1
2
-
-
1
2
3
-
-
-
2
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
726694
Liu
Cloning, expression, and chara ...
Pseudomonas sp., Pseudomonas sp. K
Appl. Biochem. Biotechnol.
166
1301-1313
2012
-
-
1
1
-
-
4
2
-
3
2
-
-
6
-
-
1
-
-
-
1
-
2
1
1
2
3
-
1
2
1
-
-
-
-
-
-
1
-
1
-
-
-
4
-
2
-
3
2
-
-
-
-
1
-
-
1
-
2
1
1
2
3
-
1
2
1
-
-
-
-
-
-
-
726981
Hung
Investigation of the role of A ...
Cupriavidus metallidurans, Cupriavidus metallidurans CH34, Methylorubrum extorquens, Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1, Nostoc punctiforme, Pseudomonas stutzeri
Biochemistry
51
4254-4262
2012
-
-
4
-
28
-
4
22
-
-
-
-
-
25
-
-
-
-
-
-
-
-
6
-
-
-
-
11
-
-
-
-
3
-
-
-
-
4
-
-
28
-
-
4
3
22
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
11
-
-
-
-
-
-
-
-
7
7
726982
Zou
Crystal structures of phosphit ...
Pseudomonas stutzeri
Biochemistry
51
4263-4270
2012
-
-
1
1
1
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
728004
Hirota
Isolation and characterization ...
Pseudomonas stutzeri, Pseudomonas stutzeri WM88, Ralstonia sp., Ralstonia sp. 4506
J. Biosci. Bioeng.
113
445-450
2012
-
-
2
-
-
-
7
8
1
-
-
-
-
11
-
-
-
-
-
-
-
-
4
-
2
-
2
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
7
-
8
1
-
-
-
-
-
-
-
-
-
-
-
4
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
691424
McLachlan
Further improvement of phosphi ...
Pseudomonas stutzeri
Biotechnol. Bioeng.
99
268-274
2008
-
1
1
-
33
-
-
10
-
-
-
-
-
1
-
-
1
-
-
-
-
1
1
-
-
-
-
4
-
-
-
1
-
-
-
-
1
1
1
-
33
-
-
-
-
10
-
-
-
-
-
-
-
1
-
-
-
1
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
672712
Johannes
Efficient regeneration of NADP ...
Pseudomonas stutzeri
Biotechnol. Bioeng.
96
18-26
2007
-
1
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
690884
Fogle
Pre-steady-state studies of ph ...
Pseudomonas stutzeri, Pseudomonas stutzeri WW88
Biochemistry
46
13101-13108
2007
-
1
1
-
6
-
1
8
-
-
-
-
-
2
-
-
1
-
-
-
-
-
2
-
-
-
-
9
-
-
-
1
-
-
-
-
1
1
1
-
6
-
-
1
-
8
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
673144
Woodyer
Optimizing a biocatalyst for i ...
Pseudomonas stutzeri
Comb. Chem. High Throughput Screen.
9
237-245
2006
-
1
-
-
7
-
-
25
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
12
-
-
-
-
-
-
-
-
1
-
-
-
7
-
-
-
-
25
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
658120
Woodyer
Site-directed mutagenesis of a ...
Pseudomonas stutzeri, Pseudomonas stutzeri WM 88
Biochemistry
44
4765-4774
2005
-
-
1
-
12
-
2
13
-
-
2
2
-
2
-
-
1
-
-
-
-
-
4
1
-
-
-
7
1
1
-
1
-
-
-
-
-
1
1
-
12
-
-
2
-
13
-
-
2
2
-
-
-
1
-
-
-
-
4
1
-
-
-
7
1
1
-
-
-
-
-
-
-
-
658126
Relyea
Inhibition and pH dependence o ...
Pseudomonas stutzeri
Biochemistry
44
6640-6649
2005
-
-
-
-
-
-
1
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
658259
Relyea
Mechanism and applications of ...
Alcaligenes faecalis, Pseudomonas stutzeri, Pseudomonas stutzeri WM88
Bioorg. Chem.
33
171-189
2005
-
1
-
-
6
-
2
-
-
-
-
3
-
5
-
-
-
1
-
-
-
-
10
-
1
-
-
-
1
-
-
3
-
-
-
-
1
-
3
-
6
-
-
2
-
-
-
-
-
3
-
-
-
-
-
-
-
-
10
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
671425
Johannes
Directed evolution of a thermo ...
Pseudomonas stutzeri
Appl. Environ. Microbiol.
71
5728-5734
2005
-
1
-
-
17
-
-
33
-
-
-
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
2
12
-
-
-
-
-
-
-
-
1
-
-
-
17
-
-
-
-
33
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
2
12
-
-
-
-
-
-
-
-
-
-
673526
Woodyer
Mechanistic investigation of a ...
Pseudomonas stutzeri
FEBS J.
272
3816-3827
2005
-
1
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
657956
Woodyer
Relaxing the nicotinamide cofa ...
Pseudomonas stutzeri
Biochemistry
42
11604-11614
2003
-
1
1
-
3
-
-
8
-
-
1
1
-
2
-
-
1
-
-
-
-
-
3
1
-
-
1
7
-
-
-
2
-
3
-
-
1
1
2
-
3
-
-
-
-
8
-
-
1
1
-
-
-
1
-
-
-
-
3
1
-
-
1
7
-
-
-
3
-
-
-
-
-
-
287853
Costas
Purification and characterizat ...
Pseudomonas stutzeri
J. Biol. Chem.
276
17429-17436
2001
-
-
1
-
-
-
9
2
-
-
1
1
-
3
-
-
1
-
-
-
1
-
4
1
-
-
-
1
1
1
-
-
-
-
-
-
-
1
-
-
-
-
-
9
-
2
-
-
1
1
-
-
-
1
-
-
1
-
4
1
-
-
-
1
1
1
-
-
-
-
-
-
-
-
287854
Vrtis
Phosphite dehydrogenase: An un ...
Pseudomonas stutzeri
J. Am. Chem. Soc.
123
2672-2673
2001
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
4
-
2
-
1
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
4
-
2
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-