Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Phosphonoacetate | substrate activation, mechanism, overview | Pseudomonas fluorescens |
Cloned (Comment) | Organism |
---|---|
recombinant expression in Escherichia coli strain JM105 | Pseudomonas fluorescens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme in in the presence of Zn2+, Mg2+, phosphonoformate and tartrate, from 30 mM K+BICINE, pH 8.5, and 1 mM ZnCl2, mixed with 10% poly(ethylene glycol) 8000, 100 mM Mg-acetate, and 5 mM phosphonoformate buffered at pH 7.0 with 100 mM MOPSO, X-ray diffraction structure determination and analysis at 2.8 A resolution, multiple isomorphous replacement with two heavy-atom derivatives | Pseudomonas fluorescens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Phosphonoformate | competitive inhibitor, the phosphonoformate ligand and two zinc ions are bound at the core domain | Pseudomonas fluorescens | |
phosphonoproprionate | competitive inhibitor | Pseudomonas fluorescens | |
Phosphonopyruvate | competitive inhibitor | Pseudomonas fluorescens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates 2fold at 3 mM | Pseudomonas fluorescens | |
additional information | no effects by Mg2+, Ca2+, Ni2+, Mn2+, Cr2+, Cu2+ at 3 mM | Pseudomonas fluorescens | |
Zn2+ | activates 10fold at 3 mM, the phosphonoformate ligand and two zinc ions are bound at the core domain | Pseudomonas fluorescens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
44239 | - |
2 * 44239, sequence calculation and gel filtration | Pseudomonas fluorescens |
93000 | - |
gel filtration, recombinant enzyme | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Phosphonoacetate + H2O | Pseudomonas fluorescens | - |
acetate + phosphate | - |
? | |
Phosphonoacetate + H2O | Pseudomonas fluorescens 23F | - |
acetate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | - |
- |
- |
Pseudomonas fluorescens 23F | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain JM105 by ammonium sulfate fractionation, and anion exchange, hydrophobic interaction, and hydroxylapatite chromatography, followed by gel filtration | Pseudomonas fluorescens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Phosphonoacetate + H2O = acetate + phosphate | structure and mechanism of the P-C bond cleaving enzyme, nucleophilic catalysis involving a zinc ion, overview. Modeling of the catalytic mechanism | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | substrate specificity, overview. No activity with fosfomycin or phosphonopyruvate | Pseudomonas fluorescens | ? | - |
? | |
additional information | substrate specificity, overview. No activity with fosfomycin or phosphonopyruvate | Pseudomonas fluorescens 23F | ? | - |
? | |
Phosphonoacetate + H2O | - |
Pseudomonas fluorescens | acetate + phosphate | - |
? | |
Phosphonoacetate + H2O | - |
Pseudomonas fluorescens 23F | acetate + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 44239, sequence calculation and gel filtration | Pseudomonas fluorescens |
More | PAc hydrolase monomer is comprised of a large alpha/beta/alpha core domain of residues 1-245 and 367-388, 6-stranded mixed beta-sheet, and a smaller alpha/beta cap domain comprising residues 255-360, as 4-stranded antiparallel beta-sheet | Pseudomonas fluorescens |
Synonyms | Comment | Organism |
---|---|---|
PAc hydrolase | - |
Pseudomonas fluorescens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 30 | assay at | Pseudomonas fluorescens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 8 | assay at | Pseudomonas fluorescens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.049 | - |
Phosphonoformate | pH 7.0, 25°C | Pseudomonas fluorescens | |
0.19 | - |
Phosphonoformate | pH 7.0, 30°C | Pseudomonas fluorescens | |
1.2 | - |
Phosphonopyruvate | pH 7.0, 30°C | Pseudomonas fluorescens | |
1.2 | - |
phosphonoproprionate | pH 7.0, 30°C | Pseudomonas fluorescens |
General Information | Comment | Organism |
---|---|---|
evolution | the P-C bond cleaving enzyme phosphonoacetate hydrolase belongs to the alkaline phosphatase superfamily | Pseudomonas fluorescens |
metabolism | the enzyme is involved in the 2-aminoethylphosphonic acid degradation pathway, overview | Pseudomonas fluorescens |