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Literature summary for 3.5.5.1 extracted from

  • Baum, S.; Williamson, D.S.; Sewell, T.; Stolz, A.
    Conversion of sterically demanding alpha,alpha-disubstituted phenylacetonitriles by the arylacetonitrilase from Pseudomonas fluorescens EBC191 (2012), Appl. Environ. Microbiol., 78, 48-57.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.5.1

Application

Application Comment Organism
synthesis recombinant cells expressing the enzyme are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones Alcaligenes faecalis
synthesis recombinant cells expressing the enzyme are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones Rhodococcus sp.
synthesis recombinant cells expressing the enzyme are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones Synechocystis sp.
synthesis recombinant cells expressing the enzyme are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones Pseudomonas fluorescens

Cloned(Commentary)

Cloned (Comment) Organism
expression under the control of a rhamnose-inducible promoter in Escherichia coli strain JM109 Alcaligenes faecalis
expression under the control of a rhamnose-inducible promoter in Escherichia coli strain JM109 Rhodococcus sp.
expression under the control of a rhamnose-inducible promoter in Escherichia coli strain JM109 Synechocystis sp.
expression under the control of a rhamnose-inducible promoter in Escherichia coli strain JM109 Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
Y54A site-directed mutagenesis, the mutant enzyme converts 2-hydroxy-2-phenylpropionitrile with about the same activity as that of the wild-type enzyme, but forms significantly reduced amounts of amides from mandelonitrile and acetophenone cyanohydrin, it shows different kinetics of acetophenone cyanohydrin conversion and product formation compared to the wild-type Pseudomonas fluorescens
Y54F site-directed mutagenesis, altered substrate specificity and enantioselectivity compared to the wild-type enzyme, overview Pseudomonas fluorescens
Y54M site-directed mutagenesis, altered substrate specificity and enantioselectivity compared to the wild-type enzyme, overview Pseudomonas fluorescens
Y54P site-directed mutagenesis, altered substrate specificity and enantioselectivity compared to the wild-type enzyme, overview Pseudomonas fluorescens
Y54V site-directed mutagenesis, altered substrate specificity and enantioselectivity compared to the wild-type enzyme, overview Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic and chiral analysis, overview Alcaligenes faecalis
additional information
-
 additional information kinetic and chiral analysis, overview Rhodococcus sp.
additional information
-
 additional information kinetic and chiral analysis, overview Synechocystis sp.
additional information
-
 additional information kinetic and chiral analysis, overview Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-methyl-2-phenylpropionitrile + 2 H2O Alcaligenes faecalis
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O Rhodococcus sp.
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O Synechocystis sp.
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O Pseudomonas fluorescens
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O Pseudomonas fluorescens EBC191
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O Alcaligenes faecalis ATCC 8750
-
 2-methyl-2-phenylpropionic acid + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Alcaligenes faecalis
-
-
-
Alcaligenes faecalis ATCC 8750
-
-
-
Pseudomonas fluorescens Q5EG61
-
-
Pseudomonas fluorescens EBC191 Q5EG61
-
-
Rhodococcus sp.
-
-
-
Synechocystis sp.
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.1
-
 enzyme in cells, substrate benzonitrile in Na/K-phosphate buffer, pH 7.0, 30°C Synechocystis sp.
0.4
-
 enzyme in cells, substrate 2-methyl-2-phenylpropionitrile in Na/K-phosphate buffer, pH 7.0, 30°C Alcaligenes faecalis
1.9
-
 enzyme in cells, substrate 2-methyl-2-phenylpropionitrile in Na/K-phosphate buffer, pH 7.0, 30°C Pseudomonas fluorescens
14.1
-
 enzyme in cells, substrate benzonitrile in Na/K-phosphate buffer, pH 7.0, 30°C Rhodococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Alcaligenes faecalis 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Rhodococcus sp. 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Synechocystis sp. 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Pseudomonas fluorescens 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Pseudomonas fluorescens EBC191 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-hydroxy-2-phenylpropionitrile + 2 H2O acetophenone cyanohydrin, the substrate is transformed in cells to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide are formed preferentially from acetophenone cyanohydrin Alcaligenes faecalis ATCC 8750 2-hydroxy-2-phenylpropionic acid + NH3 i.e. atrolactic acid ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Alcaligenes faecalis 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Rhodococcus sp. 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Synechocystis sp. 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Pseudomonas fluorescens 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Alcaligenes faecalis 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Rhodococcus sp. 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Synechocystis sp. 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Pseudomonas fluorescens 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Pseudomonas fluorescens EBC191 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Pseudomonas fluorescens EBC191 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O
-
 Alcaligenes faecalis ATCC 8750 2-methyl-2-phenylpropionic acid + NH3
-
 ?
2-methyl-2-phenylpropionitrile + 2 H2O substrate contains a quaternary carbon atom in the alpha-position toward the nitrile group Alcaligenes faecalis ATCC 8750 2-methyl-2-phenylpropionic acid + NH3
-
 ?
benzonitrile + 2 H2O
-
 Alcaligenes faecalis benzoate + NH3
-
 ?
benzonitrile + 2 H2O
-
 Rhodococcus sp. benzoate + NH3
-
 ?
benzonitrile + 2 H2O
-
 Synechocystis sp. benzoate + NH3
-
 ?
benzonitrile + 2 H2O
-
 Alcaligenes faecalis ATCC 8750 benzoate + NH3
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Alcaligenes faecalis ?
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Rhodococcus sp. ?
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Synechocystis sp. ?
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Pseudomonas fluorescens ?
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Pseudomonas fluorescens EBC191 ?
-
 ?
additional information the enzyme catalyzes the reactions of nitrilase, EC 3.5.5.1, and arylacetonitrilase, EC 3.5.5.5, substrate specificity and enantioselectivity, overview. Steric hindrance with amino acid residue Tyr54 impairs the binding or conversion of sterically demanding substrates, homology modelling Alcaligenes faecalis ATCC 8750 ?
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Alcaligenes faecalis
30
-
 assay at Rhodococcus sp.
30
-
 assay at Synechocystis sp.
30
-
 assay at Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 with substrate 2-methyl-2-phenylpropionitrile in Na/K-phosphate buffer Alcaligenes faecalis
7
-
 with substrate 2-methyl-2-phenylpropionitrile in Na/K-phosphate buffer Pseudomonas fluorescens
7
-
 with substrate benzonitrile in Na/K-phosphate buffer Rhodococcus sp.
7
-
 with substrate benzonitrile in Na/K-phosphate buffer Synechocystis sp.

General Information

General Information Comment Organism
additional information generation of a model of the nitrilase by homology modeling, overview Alcaligenes faecalis
additional information generation of a model of the nitrilase by homology modeling, overview Rhodococcus sp.
additional information generation of a model of the nitrilase by homology modeling, overview Synechocystis sp.
additional information generation of a model of the nitrilase by homology modeling, overview Pseudomonas fluorescens