Literature summary for 3.11.1.1 extracted from

Zhang, G.; Morais, M.C.; Dai, J.; Zhang, W.; Dunaway-Mariano, D.; Allen, K.N.
Investigation of metal ion binding in phosphonoacetaldehyde hydrolase identifies sequence markers for metal-activated enzymes of the HAD enzyme superfamily (2004), Biochemistry, 43, 4990-4997.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli	Bacillus cereus

Crystallization (Commentary)

Crystallization (Comment)	Organism
10 mg/ml wild-type and mutant D12A enzymes complexed with Mg2+ only or with Mg2+ and substrate, in 1 mM HEPES, 10 mM MgCl2, 0.1 mM DTT, pH 7.5, 4°C, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, the latter containing 30% PEG 4000, 100 mM Tris-HCl, pH 7.4, 100 mM MgCl2, 1 week, against the reservoir well solution additionally with 20% glycerol before data collection, X-ray diffraction structure determination and analysis at 2.3-2.55 A	Bacillus cereus

Crystallization (Comment)

Organism

10 mg/ml wild-type and mutant D12A enzymes complexed with Mg2+ only or with Mg2+ and substrate, in 1 mM HEPES, 10 mM MgCl2, 0.1 mM DTT, pH 7.5, 4°C, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, the latter containing 30% PEG 4000, 100 mM Tris-HCl, pH 7.4, 100 mM MgCl2, 1 week, against the reservoir well solution additionally with 20% glycerol before data collection, X-ray diffraction structure determination and analysis at 2.3-2.55 A

Bacillus cereus

Protein Variants

Protein Variants	Comment	Organism
D12A	site-directed mutagenesis, catalytically inactive mutant	Bacillus cereus
D186A	site-directed mutagenesis, highly reduced activity	Bacillus cereus
D186A/D190A	site-directed mutagenesis, inactive mutant	Bacillus cereus
D186E	site-directed mutagenesis, very highly reduced activity	Bacillus cereus
D190A	site-directed mutagenesis, reduced activity	Bacillus cereus
G185D/D190G	site-directed mutagenesis, reduced activity	Bacillus cereus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	kinetics, activity of mutant D186A is too low to measure Km accurately	Bacillus cereus
0.033	-	phosphonoacetaldehyde	wild-type enzyme, pH 7.5, 25°C	Bacillus cereus
0.054	-	phosphonoacetaldehyde	mutant G185D/D190G, pH 7.5, 25°C	Bacillus cereus
0.072	-	phosphonoacetaldehyde	mutant D12E, pH 7.5, 25°C	Bacillus cereus
0.52	-	phosphonoacetaldehyde	mutant D190A, pH 7.5, 25°C	Bacillus cereus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	activates, required for catalysis, serves as a cofactor, binds via ligation to the loop 1 Asp12 carboxylate and Thr 14 backbone carbonyl and to the loop 4 Asp186carboxylate, the loop 4 Asp190 forms a hydrogen bond to the Mg(II) water ligand, Asp186 is essential while Asp190 simply enhances cofactor binding	Bacillus cereus

Organism

Organism	UniProt	Comment	Textmining
Bacillus cereus	O31156	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate	active site structure	Bacillus cereus	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
phosphonoacetaldehyde + H2O	-	Bacillus cereus	acetaldehyde + phosphate	-	ir

Synonyms

Synonyms	Comment	Organism
phosphonatase	-	Bacillus cereus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assy at	Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.000084	-	phosphonoacetaldehyde	mutant D186A, pH 7.5, 25°C	Bacillus cereus
0.0012	-	phosphonoacetaldehyde	mutant D12E, pH 7.5, 25°C	Bacillus cereus
0.022	-	phosphonoacetaldehyde	mutant D190A, pH 7.5, 25°C	Bacillus cereus
1.7	-	phosphonoacetaldehyde	mutant G185D/D190G, pH 7.5, 25°C	Bacillus cereus
15	-	phosphonoacetaldehyde	wild-type enzyme, pH 7.5, 25°C	Bacillus cereus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Bacillus cereus