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Literature summary extracted from

  • Yeom, S.J.; Lee, J.K.; Oh, D.K.
    A positively charged amino acid at position 129 in nitrilase from Rhodococcus rhodochrous ATCC 33278 is an essential residue for the activity with meta-substituted benzonitriles (2010), FEBS Lett., 584, 106-110.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.1 expressed in Escherichia coli strain ER2566 Rhodococcus rhodochrous

Protein Variants

EC Number Protein Variants Comment Organism
3.5.5.1 G103A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 I104A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 M114A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 N164A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 Q116A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 R129A the mutant exhibits no activity with 3-methylbenzonitrile and benzonitrile Rhodococcus rhodochrous
3.5.5.1 R129H the mutant enzyme is active only for meta-substituted benzonitriles Rhodococcus rhodochrous
3.5.5.1 R129K the mutant enzyme is active only for meta-substituted benzonitriles Rhodococcus rhodochrous
3.5.5.1 R130A the mutant exhibits increased specific activity with 3-methylbenzonitrile and benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 S188A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 T115A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 V49A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous
3.5.5.1 Y142A the mutant exhibits decreased specific activity with 3-methylbenzonitrile and increased specific activity with benzonitrile compared to the wild type enzyme Rhodococcus rhodochrous

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.1 0.456
-
 3-aminobenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.483
-
 3-aminobenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.55
-
 3-methylbenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.592
-
 3-methylbenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 1.068
-
 Benzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 1.468
-
 Benzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 1.699
-
 4-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 1.851
-
 3-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 1.863
-
 3-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 2.426
-
 4-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 3.13
-
 Benzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.1 Rhodococcus rhodochrous
-
-
-
3.5.5.1 Rhodococcus rhodochrous ATCC 33278
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.5.1 0.6
-
 mutant enzyme N164A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 5.1
-
 mutant enzyme V49A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 5.2
-
 mutant enzyme M114A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 7
-
 wild type enzyme, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 7.5
-
 mutant enzyme T115A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 8
-
 mutant enzyme S188A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 8.2
-
 mutant enzyme T115A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 8.8
-
 mutant enzyme G103A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 8.8
-
 mutant enzyme I104A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 9
-
 mutant enzyme Q116A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 9.6
-
 mutant enzyme G103A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 9.7
-
 mutant enzyme N164A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 9.9
-
 mutant enzyme M114A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 9.9
-
 mutant enzyme Q116A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 10
-
 mutant enzyme S188A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 10.4
-
 mutant enzyme I104A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 10.7
-
 wild type enzyme, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 11.6
-
 mutant enzymeV49A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 14.5
-
 mutant enzyme R130A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 17.7
-
 mutant enzyme R130A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 22.4
-
 mutant enzyme Y142A, using benzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 26
-
 mutant enzyme Y142A, using 3-methylbenzonitrile as substrate, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.1 3-aminobenzonitrile + H2O
-
 Rhodococcus rhodochrous 3-aminobenzoic acid + NH3
-
 ?
3.5.5.1 3-aminobenzonitrile + H2O
-
 Rhodococcus rhodochrous ATCC 33278 3-aminobenzoic acid + NH3
-
 ?
3.5.5.1 3-methylbenzonitrile + H2O
-
 Rhodococcus rhodochrous 3-methylbenzoic acid + NH3
-
 ?
3.5.5.1 3-methylbenzonitrile + H2O
-
 Rhodococcus rhodochrous ATCC 33278 3-methylbenzoic acid + NH3
-
 ?
3.5.5.1 4-aminobenzonitrile + H2O
-
 Rhodococcus rhodochrous 4-aminobenzoic acid + NH3
-
 ?
3.5.5.1 4-aminobenzonitrile + H2O
-
 Rhodococcus rhodochrous ATCC 33278 4-aminobenzoic acid + NH3
-
 ?
3.5.5.1 4-methylbenzonitrile + H2O
-
 Rhodococcus rhodochrous 4-methylbenzoic acid + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O
-
 Rhodococcus rhodochrous benzoic acid + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O
-
 Rhodococcus rhodochrous ATCC 33278 benzoic acid + NH3
-
 ?
3.5.5.1 additional information the wild type enzyme containing Arg129 is active only for meta- and para-substituted benzonitriles with a methyl or amino group, but shows no activity for ortho-substituted benzonitriles Rhodococcus rhodochrous ?
-
 ?
3.5.5.1 additional information the wild type enzyme containing Arg129 is active only for meta- and para-substituted benzonitriles with a methyl or amino group, but shows no activity for ortho-substituted benzonitriles Rhodococcus rhodochrous ATCC 33278 ?
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.5.1 0.000129
-
 Benzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.00014
-
 3-aminobenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.00015
-
 3-aminobenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000158
-
 3-methylbenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000162
-
 Benzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.00018
-
 3-methylbenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000198
-
 3-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000283
-
 4-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000288
-
 3-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.00047
-
 Benzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.000695
-
 4-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.5.1 0.001068
-
 3-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.0011
-
 Benzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.001223
-
 Benzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.0015
-
 Benzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.001548
-
 3-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.001667
-
 4-aminobenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.002867
-
 4-methylbenzonitrile wild type enzyme, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.0029
-
 3-methylbenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.003
-
 3-methylbenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.00308
-
 3-aminobenzonitrile mutant enzyme R129K, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous
3.5.5.1 0.0032
-
 3-aminobenzonitrile mutant enzyme R129H, in 50 mM potassium phosphate (pH 7.5), at 25°C Rhodococcus rhodochrous