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Literature summary for 1.20.1.1 extracted from

  • Hung, J.E.; Fogle, E.J.; Garg, N.; Chekan, J.R.; Nair, S.K.; van der Donk, W.A.
    Chemical rescue and inhibition studies to determine the role of Arg301 in phosphite dehydrogenase (2014), PLoS ONE, 9, e87134 .
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) 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

Protein Variants

Protein Variants Comment 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 Comment Organism Structure
sulfite competitive inhibitor Pseudomonas stutzeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphonate + NAD+ + H2O Pseudomonas stutzeri
-
phosphate + NADH + H+
-
?
phosphonate + NAD+ + H2O Pseudomonas stutzeri WM88
-
phosphate + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas stutzeri O69054
-
-
Pseudomonas stutzeri WM88 O69054
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-affinity column chromatography and Superdex 200 gel filtration Pseudomonas stutzeri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
phosphonate + NAD+ + H2O
-
Pseudomonas stutzeri phosphate + NADH + H+
-
?
phosphonate + NAD+ + H2O
-
Pseudomonas stutzeri WM88 phosphate + NADH + H+
-
?

Synonyms

Synonyms Comment Organism
phosphite dehydrogenase
-
Pseudomonas stutzeri
PTDH
-
Pseudomonas stutzeri

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pseudomonas stutzeri

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment 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