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Literature summary for 1.2.1.3 extracted from

  • Agarwal, V.; Peck, S.C.; Chen, J.H.; Borisova, S.A.; Chekan, J.R.; van der Donk, W.A.; Nair, S.K.
    Structure and function of phosphonoacetaldehyde dehydrogenase: the missing link in phosphonoacetate formation (2014), Chem. Biol., 21, 125-135.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.2.1.3

Activating Compound

Activating Compound Comment Organism Structure
ascorbate required Sinorhizobium meliloti

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta2 cells Sinorhizobium meliloti

Crystallization (Commentary)

Crystallization (Comment) Organism
apoenzyme and in complex with PnA und NAD+, hanging drop vapor diffusion method, using 22%-25% PEG3350, 0.1 M sodium cacodylate (pH 6.5), 0.2 M sodium phosphate dibasic, and 5%-10% glycerol Sinorhizobium meliloti

Protein Variants

Protein Variants Comment Organism
E385A the mutation results in 10fold reduction in kcat, with an approximately 6fold higher KM,NAD+ Sinorhizobium meliloti
N158A the mutation results in a 200fold decrease in kcat Sinorhizobium meliloti
R108A the mutation leads to a 40fold decrease in kcat and a 3fold increase in KM,PnAA Sinorhizobium meliloti
R290A the mutation results in a 20fold decrease in kcat with a slightly elevated KM,PnAA relative to wild type Sinorhizobium meliloti
R447A the mutation reduces the kcat 30fold and increases the KM,PnAA 50fold Sinorhizobium meliloti

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0032
-
 phosphonoacetaldehyde wild type enzyme, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.005
-
 phosphonoacetaldehyde mutant enzyme R290A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.009
-
 phosphonoacetaldehyde mutant enzyme R108A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.018
-
 NAD+ mutant enzyme R108A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.019
-
 phosphonoacetaldehyde mutant enzyme E385A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.029
-
 phosphonoacetaldehyde mutant enzyme N158A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.04
-
 NAD+ mutant enzyme N158A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.054
-
 NAD+ mutant enzyme R447A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.058
-
 NAD+ wild type enzyme, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.075
-
 NAD+ mutant enzyme R290A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.15
-
 phosphonoacetaldehyde mutant enzyme R447A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.37
-
 NAD+ mutant enzyme E385A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ required Sinorhizobium meliloti

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphonoacetaldehyde + NAD+ + H2O Sinorhizobium meliloti
-
 phosphonoacetate + NADH + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Sinorhizobium meliloti Q92UV7
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA resin column chromatography and Superdex 75 gel filtration Sinorhizobium meliloti

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-oxopropylphosphonate + NAD+ + H2O
-
 Sinorhizobium meliloti ?
-
 ?
glyceraldehyde-3-phosphate + NAD+ + H2O
-
 Sinorhizobium meliloti ?
-
 ?
phosphonoacetaldehyde + NAD+ + H2O
-
 Sinorhizobium meliloti phosphonoacetate + NADH + H+
-
 ?

Synonyms

Synonyms Comment Organism
phnY
-
 Sinorhizobium meliloti
phosphonoacetaldehyde oxidase
-
 Sinorhizobium meliloti

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 phosphonoacetaldehyde mutant enzyme N158A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.051
-
 phosphonoacetaldehyde mutant enzyme R108A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.076
-
 phosphonoacetaldehyde mutant enzyme R447A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.12
-
 phosphonoacetaldehyde mutant enzyme R290A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
0.19
-
 phosphonoacetaldehyde mutant enzyme E385A, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti
2.2
-
 phosphonoacetaldehyde wild type enzyme, with NAD+ in 50 mM HEPES, pH 7.5 supplemented with 10 mM MgSO4 for 5 min at 30°C Sinorhizobium meliloti

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Sinorhizobium meliloti