Activating Compound | Comment | Organism | Structure |
---|---|---|---|
ascorbate | required | Sinorhizobium meliloti |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta2 cells | Sinorhizobium meliloti |
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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | required | Sinorhizobium meliloti |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphonoacetaldehyde + NAD+ + H2O | Sinorhizobium meliloti | - |
phosphonoacetate + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sinorhizobium meliloti | Q92UV7 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA resin column chromatography and Superdex 75 gel filtration | Sinorhizobium meliloti |
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 | Comment | Organism |
---|---|---|
phnY | - |
Sinorhizobium meliloti |
phosphonoacetaldehyde oxidase | - |
Sinorhizobium meliloti |
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 | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Sinorhizobium meliloti |