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Literature summary extracted from

  • Gopalakrishna, K.N.; Stewart, B.H.; Kneen, M.M.; Andricopulo, A.D.; Kenyon, G.L.; McLeish, M.J.
    Mandelamide hydrolase from Pseudomonas putida: characterization of a new member of the amidase signature family (2004), Biochemistry, 43, 7725-7735.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.86 expression in Escherichia coli both wild type and His6-tagged protein Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.86 K100A no enzymic activity Pseudomonas putida
3.5.1.86 S180A significant decrease in kcat value Pseudomonas putida
3.5.1.86 S181A significant decrease in kcat value Pseudomonas putida
3.5.1.86 S204A no enzymic activity Pseudomonas putida
3.5.1.86 S217A very little enzymic activity Pseudomonas putida
3.5.1.86 S218A very little enzymic activity Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.86 3-phenyl-1,1,1-trifluoro-2-propanone competitive Pseudomonas putida
3.5.1.86 3-phenyl-1-chloro-2-propanone competitive Pseudomonas putida
3.5.1.86 PMSF 1 mM, 80% inhibition Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.86 0.0038
-
 2-phenylacetamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.009
-
 (S)-mandelamide pH 7.5, 30°C, mutant S181A Pseudomonas putida
3.5.1.86 0.02
-
 (S)-mandelamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.032
-
 N-methyl phenylacetamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.033
-
 (R)-mandelamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.049
-
 3-phenylpropionamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.059
-
 (R)-mandelamide pH 7.5, 30°C, mutant S181A Pseudomonas putida
3.5.1.86 0.17
-
 (R,S)-methyl mandelate pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.288
-
 hexanoamide pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.296
-
 (S)-mandelamide pH 7.5, 30°C, mutant S180A Pseudomonas putida
3.5.1.86 0.871
-
 (R)-mandelamide pH 7.5, 30°C, mutant S180A Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.86 53800
-
 x * 54000, SDS-PAGE, x * 53800, mass spectrometry Pseudomonas putida
3.5.1.86 54000
-
 x * 54000, SDS-PAGE, x * 53800, mass spectrometry Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.86 Pseudomonas putida Q84DC4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.86 recombinant enzyme Pseudomonas putida

Storage Stability

EC Number Storage Stability Organism
3.5.1.86 -20°C, no loss of activtiy during several freeze-thaw cycles Pseudomonas putida
3.5.1.86 4°C, phosphate buffer pH 7.8, 1 mM EDTA, stable for at least 1 month Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.86 (R)-mandelamide + H2O
-
 Pseudomonas putida (R)-mandelate + NH3
-
 ?
3.5.1.86 (R,S)-methyl mandelate + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 (S)-mandelamide + H2O
-
 Pseudomonas putida (S)-mandelate + NH3
-
 ?
3.5.1.86 2-phenylacetamide + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 3-phenylpropionamide + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 hexanoamide + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 N-methyl phenylacetamide + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 R,S-mandelamide + H2O
-
 Pseudomonas putida mandelic acid + ammonia
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.86 ? x * 54000, SDS-PAGE, x * 53800, mass spectrometry Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
3.5.1.86 MAH
-
 Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.86 7.8
-
-
 Pseudomonas putida

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.5.1.86 0.06
-
 3-phenyl-1,1,1-trifluoro-2-propanone pH 7.5, 30°C Pseudomonas putida
3.5.1.86 0.13
-
 3-phenyl-1-chloro-2-propanone pH 7.5, 30°C Pseudomonas putida