Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary 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.
    View publication on PubMed

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.11.1.1 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.11.1.1 additional information
-
 additional information kinetics, activity of mutant D186A is too low to measure Km accurately Bacillus cereus
3.11.1.1 0.033
-
 phosphonoacetaldehyde wild-type enzyme, pH 7.5, 25°C Bacillus cereus
3.11.1.1 0.054
-
 phosphonoacetaldehyde mutant G185D/D190G, pH 7.5, 25°C Bacillus cereus
3.11.1.1 0.072
-
 phosphonoacetaldehyde mutant D12E, pH 7.5, 25°C Bacillus cereus
3.11.1.1 0.52
-
 phosphonoacetaldehyde mutant D190A, pH 7.5, 25°C Bacillus cereus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.11.1.1 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

EC Number Organism UniProt Comment Textmining
3.11.1.1 Bacillus cereus O31156
-
-

Reaction

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

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
3.11.1.1 phosphonatase
-
 Bacillus cereus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.11.1.1 25
-
 assy at Bacillus cereus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.11.1.1 0.000084
-
 phosphonoacetaldehyde mutant D186A, pH 7.5, 25°C Bacillus cereus
3.11.1.1 0.0012
-
 phosphonoacetaldehyde mutant D12E, pH 7.5, 25°C Bacillus cereus
3.11.1.1 0.022
-
 phosphonoacetaldehyde mutant D190A, pH 7.5, 25°C Bacillus cereus
3.11.1.1 1.7
-
 phosphonoacetaldehyde mutant G185D/D190G, pH 7.5, 25°C Bacillus cereus
3.11.1.1 15
-
 phosphonoacetaldehyde wild-type enzyme, pH 7.5, 25°C Bacillus cereus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.11.1.1 7.5
-
 assay at Bacillus cereus