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Literature summary for 1.2.1.3 extracted from

  • van Staalduinen, L.M.; McSorley, F.R.; Schiessl, K.; Seguin, J.; Wyatt, P.B.; Hammerschmidt, F.; Zechel, D.L.; Jia, Z.
    Crystal structure of PhnZ in complex with substrate reveals a di-iron oxygenase mechanism for catabolism of organophosphonates (2014), Proc. Natl. Acad. Sci. USA, 111, 5171-5176 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.2.1.3

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 22%-25% (w/v) PEG3350, 0.1 M sodium cacodylate (pH 6.5), 0.2 M sodium phosphate dibasic, and 5%-10% (v/v) glycerol Sinorhizobium meliloti

Protein Variants

Protein Variants Comment Organism
C291A inactive Sinorhizobium meliloti
E254A inactive Sinorhizobium meliloti
E385A the mutant shows strongly reduced activity compared to the wild type enzyme Sinorhizobium meliloti
N158A the mutant shows strongly reduced activity compared to the wild type enzyme Sinorhizobium meliloti
R108A the mutant shows strongly reduced activity compared to the wild type enzyme Sinorhizobium meliloti
R290A the mutant shows strongly reduced activity compared to the wild type enzyme Sinorhizobium meliloti
R447A the mutant shows strongly reduced activity compared to the wild type enzyme Sinorhizobium meliloti

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0032
-
 phosphonoacetaldehyde wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti
0.0051
-
 phosphonoacetaldehyde mutant enzyme R290A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.0097
-
 phosphonoacetaldehyde mutant enzyme R108A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.018
-
 NAD+ mutant enzyme R108A, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.019
-
 phosphonoacetaldehyde mutant enzyme E385A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.029
-
 phosphonoacetaldehyde mutant enzyme N158A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.04
-
 NAD+ mutant enzyme N158A, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.054
-
 NAD+ mutant enzyme R447A, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.058
-
 NAD+ wild type enzyme, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.075
-
 NAD+ mutant enzyme R290A, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.097
-
 glyceraldehyde 3-phosphate wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti
0.15
-
 phosphonoacetaldehyde mutant enzyme R447A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.37
-
 NAD+ mutant enzyme E385A, with phosphonoacetaldehyde as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
0.53
-
 NAD+ wild type enzyme, with glyceraldehyde-3-phosphate as cosubstrate, at pH 7.5 and 30°C Sinorhizobium meliloti
3.3
-
 3-oxopropyl phosphonate wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti

Organism

Organism UniProt Comment Textmining
Sinorhizobium meliloti Q92UV7
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-oxopropyl phosphonate + 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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 phosphonoacetaldehyde mutant enzyme N158A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.051
-
 phosphonoacetaldehyde mutant enzyme R108A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.076
-
 phosphonoacetaldehyde mutant enzyme R447A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.098
-
 glyceraldehyde 3-phosphate wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti
0.12
-
 phosphonoacetaldehyde mutant enzyme R290A, at pH 7.5 and 30°C Sinorhizobium meliloti
0.19
-
 phosphonoacetaldehyde mutant enzyme E385A, at pH 7.5 and 30°C Sinorhizobium meliloti
1.5
-
 3-oxopropyl phosphonate wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti
2.2
-
 phosphonoacetaldehyde wild type enzyme, at pH 7.5 and 30°C Sinorhizobium meliloti

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Sinorhizobium meliloti