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Literature summary for 1.20.1.1 extracted from

  • Woodyer, R.; Wheatley, J.L.; Relyea, H.A.; Rimkus, S.; van der Donk, W.A.
    Site-directed mutagenesis of active site residues of phosphite dehydrogenase (2005), Biochemistry, 44, 4765-4774.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
wild type and mutant proteins expressed in Escherichia coli BL21(DE3) as His-tag fusion protein Pseudomonas stutzeri

Protein Variants

Protein Variants Comment Organism
E266Q significant increase in Km for both substrates, increase in turnover Pseudomonas stutzeri
H292F almost complete loss of activity Pseudomonas stutzeri
H292K almost complete loss of activity Pseudomonas stutzeri
H292N almost complete loss of activity Pseudomonas stutzeri
K76A significant increase in Km for both substrates Pseudomonas stutzeri
K76C significant increase in Km for both substrates Pseudomonas stutzeri
K76M significant increase in Km for both substrates Pseudomonas stutzeri
K76R significant increase in Km for both substrates Pseudomonas stutzeri
R237H almost complete loss of activity Pseudomonas stutzeri
R237K significant increase in Km for both substrates Pseudomonas stutzeri
R237L almost complete loss of activity Pseudomonas stutzeri
R237Q almost complete loss of activity Pseudomonas stutzeri

Inhibitors

Inhibitors Comment Organism Structure
2-Bromoethylamine 1 mM NAD+ protects against inactivation Pseudomonas stutzeri
diethyl dicarbonate
-
Pseudomonas stutzeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.053
-
phosphonate pH 7.5, 25°C, wild type enzyme Pseudomonas stutzeri
0.057
-
NAD+ pH 7.5, 25°C, wild type enzyme Pseudomonas stutzeri
0.11
-
NAD+ pH 7.5, 25°C, K76R mutant enzyme Pseudomonas stutzeri
0.2
-
NAD+ pH 7.5, 25°C, K76C mutant enzyme Pseudomonas stutzeri
0.26
-
NAD+ pH 7.5, 25°C, K76A mutant enzyme Pseudomonas stutzeri
0.35
-
phosphonate pH 7.5, 25°C, K76R mutant enzyme Pseudomonas stutzeri
0.6
-
phosphonate pH 7.5, 25°C, K76C mutant enzyme Pseudomonas stutzeri
0.9
-
NAD+ pH 7.5, 25°C, K76M mutant enzyme Pseudomonas stutzeri
1
-
NAD+ pH 7.5, 25°C, E266Q mutant enzyme Pseudomonas stutzeri
1
-
NAD+ pH 7.5, 25°C, R237K mutant enzyme Pseudomonas stutzeri
1.1
-
phosphonate pH 7.5, 25°C, K76A mutant enzyme Pseudomonas stutzeri
2
-
phosphonate pH 7.5, 25°C, K76M mutant enzyme Pseudomonas stutzeri
5
-
phosphonate pH 7.5, 25°C, E266Q mutant enzyme Pseudomonas stutzeri
6
-
phosphonate pH 7.5, 25°C, R237K mutant enzyme Pseudomonas stutzeri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36400
-
x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein Pseudomonas stutzeri
38500
-
x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein Pseudomonas stutzeri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphonate + H2O + NAD+ Pseudomonas stutzeri NADH regeneration system phosphate + NADH
-
ir
phosphonate + H2O + NAD+ Pseudomonas stutzeri WM 88 NADH regeneration system phosphate + NADH
-
ir

Organism

Organism UniProt Comment Textmining
Pseudomonas stutzeri
-
-
-
Pseudomonas stutzeri WM 88
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzymes using His-tag Pseudomonas stutzeri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
phosphonate + H2O + NAD+
-
Pseudomonas stutzeri phosphate + NADH
-
ir
phosphonate + H2O + NAD+ NADH regeneration system Pseudomonas stutzeri phosphate + NADH
-
ir
phosphonate + H2O + NAD+
-
Pseudomonas stutzeri WM 88 phosphate + NADH
-
ir
phosphonate + H2O + NAD+ NADH regeneration system Pseudomonas stutzeri WM 88 phosphate + NADH
-
ir

Subunits

Subunits Comment Organism
? x * 38500, SDS-PAGE, His-tag fusion protein, x * 36400, SDS-PAGE, native protein Pseudomonas stutzeri

Synonyms

Synonyms Comment Organism
PTDH
-
Pseudomonas stutzeri

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.28
-
phosphonate pH 7.5, 25°C, R237K mutant enzyme Pseudomonas stutzeri
0.58
-
phosphonate pH 7.5, 25°C, K76C mutant enzyme Pseudomonas stutzeri
0.82
-
phosphonate pH 7.5, 25°C, K76A mutant enzyme Pseudomonas stutzeri
1.62
-
phosphonate pH 7.5, 25°C, K76M mutant enzyme Pseudomonas stutzeri
2.33
-
phosphonate pH 7.5, 25°C, K76R mutant enzyme Pseudomonas stutzeri
3.2
-
phosphonate pH 7.5, 25°C, wild type enzyme Pseudomonas stutzeri
8
-
phosphonate pH 7.5, 25°C, E266Q mutant enzyme Pseudomonas stutzeri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Pseudomonas stutzeri

pH Range

pH Minimum pH Maximum Comment Organism
6 9.5
-
Pseudomonas stutzeri

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pseudomonas stutzeri