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Literature summary for 3.5.1.86 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
Search for more literature for 3.5.1.86

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Pseudomonas putida Q84DC4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Pseudomonas putida

Storage Stability

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
MAH
-
 Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
-
 Pseudomonas putida

Ki Value [mM]

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