Literature summary for 3.11.1.2 extracted from

Kim, A.; Benning, M.M.; OkLee, S.; Quinn, J.; Martin, B.M.; Holden, H.M.; Dunaway-Mariano, D.
Divergence of chemical function in the alkaline phosphatase superfamily: structure and mechanism of the P-C bond cleaving enzyme phosphonoacetate hydrolase (2011), Biochemistry, 50, 3481-3494.

Activating Compound

Activating Compound	Comment	Organism	Structure
Phosphonoacetate	substrate activation, mechanism, overview	Pseudomonas fluorescens

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression in Escherichia coli strain JM105	Pseudomonas fluorescens

Crystallization (Commentary)

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

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

Inhibitors	Comment	Organism
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

Metals/Ions	Comment	Organism
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 [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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Pseudomonas fluorescens	-	-	-
Pseudomonas fluorescens 23F	-	-	-

Purification (Commentary)

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

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	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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

Synonyms	Comment	Organism
PAc hydrolase	-	Pseudomonas fluorescens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	30	assay at	Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	8	assay at	Pseudomonas fluorescens

Ki Value [mM]

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

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