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Literature summary for 3.5.5.5 extracted from

  • Layh, N.; Parratt, J.; Willetts, A.
    Characterization and partial purification of an enantioselective arylacetonitrilase from Pseudomonas fluorescens DSM 7155 (1998), J. Mol. Catal. B, 5, 467-474.
No PubMed abstract available
Search for more literature for 3.5.5.5

Activating Compound

Activating Compound Comment Organism Structure
2-mercaptoethanol 1.4fold at 50 mM, 1.6fold activation at 100 mM Pseudomonas fluorescens
phenylacetonitrile absolutely required as growth nitrogen source substrate for induction of the enzyme, cannot be substituted by ammonia or epsilon-caprolactam Pseudomonas fluorescens

Inhibitors

Inhibitors Comment Organism Structure
Ag+ complete inhibition at 1 mM Pseudomonas fluorescens
Cu2+ complete inhibition at 1 mM Pseudomonas fluorescens
Fe2+ complete inhibition at 10 mM Pseudomonas fluorescens
Fe3+ complete inhibition at 1 mM Pseudomonas fluorescens
additional information no inhibition by KCN, EDTA, D-cycloserine, and Mn2+ Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.065
-
 phenylacetonitrile pH 7.4, 30°C Pseudomonas fluorescens
0.087
-
 rac 2-(methoxy)-mandelonitrile pH 7.4, 30°C Pseudomonas fluorescens

Metals/Ions

Metals/Ions Comment Organism Structure
additional information enzyme does not seem to require metals for activity Pseudomonas fluorescens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
13000
-
 gel filtration Pseudomonas fluorescens
38000
-
 1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE Pseudomonas fluorescens
40000
-
 1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE Pseudomonas fluorescens
57000
-
 1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE Pseudomonas fluorescens

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens
-
 DSM 7155, inducible enzyme
-

Purification (Commentary)

Purification (Comment) Organism
partial, 259fold Pseudomonas fluorescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
90
-
 partially purified enzyme Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-phenylpropionitrile + H2O low activity, 2% activity compared to phenylacetonitrile Pseudomonas fluorescens 2-phenylpropionate + NH3
-
 ?
additional information no activity with benzonitrile, 2-cyanotoluene, 3-cyanotoluene, 3-cyanobenzothiophene, propionitrile, butyronitrile, and valeronitrile, 2-phenylbutyronitrile is a poor substrate, enzyme is enantioselective and specific for arylacetonitriles Pseudomonas fluorescens ?
-
 ?
phenylacetonitrile + 2 H2O best substrate Pseudomonas fluorescens phenylacetate + NH3
-
 ?
rac 2-(methoxy)-mandelonitrile + H2O
-
 Pseudomonas fluorescens (R)-2-(methoxy)-mandelic acid + (S)-2-(methoxy)-mandelic acid + NH3 10% activity compared to phenylacetonitrile, 92% of the product is the (R)-enantiomer when the overall conversion rate is 50%, at 85% overall conversion, the level of formed (R)-enantiomer is 27% ?

Subunits

Subunits Comment Organism
trimer 1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE Pseudomonas fluorescens

Synonyms

Synonyms Comment Organism
nitrilase
-
 Pseudomonas fluorescens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
 Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Pseudomonas fluorescens