BRENDA - Enzyme Database show
show all sequences of 3.5.5.5

Characterization and partial purification of an enantioselective arylacetonitrilase from Pseudomonas fluorescens DSM 7155

Layh, N.; Parratt, J.; Willetts, A.; J. Mol. Catal. B 5, 467-474 (1998)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
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
Commentary
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
Commentary
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
Commentary
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]
Commentary
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
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas fluorescens
-
DSM 7155, inducible enzyme
-
Purification (Commentary)
Commentary
Organism
partial, 259fold
Pseudomonas fluorescens
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
90
-
partially purified enzyme
Pseudomonas fluorescens
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-phenylpropionitrile + H2O
low activity, 2% activity compared to phenylacetonitrile
652959
Pseudomonas fluorescens
2-phenylpropionate + NH3
-
652959
Pseudomonas fluorescens
?
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
652959
Pseudomonas fluorescens
?
-
652959
Pseudomonas fluorescens
?
phenylacetonitrile + H2O
best substrate
652959
Pseudomonas fluorescens
phenylacetate + NH3
-
652959
Pseudomonas fluorescens
?
rac 2-(methoxy)-mandelonitrile + H2O
-
652959
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%
652959
Pseudomonas fluorescens
?
Subunits
Subunits
Commentary
Organism
trimer
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
Pseudomonas fluorescens
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
55
-
-
Pseudomonas fluorescens
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
-
Pseudomonas fluorescens
Activating Compound (protein specific)
Activating Compound
Commentary
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 (protein specific)
Inhibitors
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
enzyme does not seem to require metals for activity
Pseudomonas fluorescens
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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
Purification (Commentary) (protein specific)
Commentary
Organism
partial, 259fold
Pseudomonas fluorescens
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
90
-
partially purified enzyme
Pseudomonas fluorescens
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-phenylpropionitrile + H2O
low activity, 2% activity compared to phenylacetonitrile
652959
Pseudomonas fluorescens
2-phenylpropionate + NH3
-
652959
Pseudomonas fluorescens
?
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
652959
Pseudomonas fluorescens
?
-
652959
Pseudomonas fluorescens
?
phenylacetonitrile + H2O
best substrate
652959
Pseudomonas fluorescens
phenylacetate + NH3
-
652959
Pseudomonas fluorescens
?
rac 2-(methoxy)-mandelonitrile + H2O
-
652959
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%
652959
Pseudomonas fluorescens
?
Subunits (protein specific)
Subunits
Commentary
Organism
trimer
1 * 57000 + 1 * 40000 + 1 * 38000, SDS-PAGE
Pseudomonas fluorescens
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
55
-
-
Pseudomonas fluorescens
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
-
Pseudomonas fluorescens
Other publictions for EC 3.5.5.5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733193
Sosedov
Improvement of the amides form ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Microbiol. Biotechnol.
99
2623-2635
2015
-
-
1
-
1
-
-
-
-
-
-
-
-
11
-
-
-
-
-
-
-
-
4
-
-
-
-
-
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-
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1
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1
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-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
733558
Sun
Cloning, purification and eval ...
Luminiphilus syltensis, Luminiphilus syltensis NOR5-1B
Biotechnol. Lett.
37
1655-1661
2015
-
-
1
-
-
-
-
1
-
-
1
-
-
4
-
-
1
-
-
-
-
-
41
1
1
1
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
1
-
-
-
-
41
1
1
1
1
-
1
-
-
-
-
-
-
-
-
-
733546
Bhatia
Purification and characterizat ...
Alcaligenes sp., Alcaligenes sp. MTCC 10675
Biotechnol. Appl. Biochem.
61
459-465
2014
-
-
-
-
-
1
10
1
-
1
2
-
-
3
-
-
1
-
-
-
-
-
8
1
1
-
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
10
-
1
-
1
2
-
-
-
-
1
-
-
-
-
8
1
1
-
1
1
1
-
-
-
-
-
-
-
-
-
718648
Baum
Conversion of sterically deman ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Environ. Microbiol.
78
48-57
2012
-
1
1
-
11
-
-
1
-
-
-
2
-
10
-
-
-
-
-
-
-
-
14
-
1
-
-
-
2
-
-
-
-
-
-
-
1
1
-
-
11
-
-
-
-
1
-
-
-
2
-
-
-
-
-
-
-
-
14
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
718670
Petrickova
Purification and characterizat ...
Aspergillus niger, Aspergillus niger CBS 513.88, Neurospora crassa, Neurospora crassa OR74A
Appl. Microbiol. Biotechnol.
93
1553-1561
2012
-
-
2
-
-
-
-
6
-
-
-
6
-
7
-
-
2
-
-
-
-
-
24
-
2
-
8
-
2
-
9
-
-
-
-
-
-
3
-
-
-
-
-
-
-
9
-
-
-
6
-
-
-
3
-
-
-
-
24
-
3
-
12
-
3
-
14
-
-
1
1
-
-
-
734496
Petrickova
-
Influence of point mutations n ...
Aspergillus niger, Aspergillus niger CBS 513.88, Neurospora crassa, Neurospora crassa DSM 1257
J. Mol. Catal. B
77
74-80
2012
-
-
2
-
4
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
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8
-
-
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-
-
2
4
-
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2
-
-
4
-
-
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-
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-
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-
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-
-
-
-
-
8
-
-
-
-
-
2
4
-
-
-
-
-
-
-
-
719297
Detzel
-
Autodisplay of nitrilase from ...
Alcaligenes faecalis, Alcaligenes faecalis ATCC 8750
ChemCatChem
3
719-725
2011
-
-
1
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
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-
-
-
-
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-
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-
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-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
719629
Liu
Gene cloning, expression, and ...
Alcaligenes faecalis, Alcaligenes faecalis ZJUTB10
J. Agric. Food Chem.
59
11560-11570
2011
-
1
1
-
3
-
11
1
-
2
1
2
-
2
-
-
1
1
-
-
-
-
12
2
1
1
1
-
1
-
-
-
-
-
-
-
1
1
-
-
3
-
-
11
-
1
-
2
1
2
-
-
-
1
-
-
-
-
12
2
1
1
1
-
1
-
-
-
-
1
1
-
-
-
710940
Sosedov
Construction and application o ...
Pseudomonas fluorescens
Appl. Environ. Microbiol.
76
3668-3674
2010
-
-
-
-
14
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
2
-
-
-
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14
-
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-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
712643
He
Biocatalytic synthesis of (R)- ...
Alcaligenes faecalis
J. Ind. Microbiol. Biotechnol.
37
741-750
2010
-
-
-
-
-
-
-
-
-
-
-
4
-
2
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5
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4
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5
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
710930
Kiziak
Identification of amino acid r ...
Pseudomonas fluorescens
Appl. Environ. Microbiol.
75
5592-5599
2009
-
-
1
-
8
-
-
-
-
-
-
-
-
5
-
-
-
-
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-
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2
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1
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8
-
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-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
712260
Thuku
Microbial nitrilases: Versatil ...
Alcaligenes faecalis, Alcaligenes faecalis ATCC 8750
J. Appl. Microbiol.
106
703-727
2009
-
-
-
-
-
-
-
-
-
-
2
-
-
4
-
-
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-
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6
1
1
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1
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2
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-
-
-
-
6
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
713073
Howden
Pseudomonas syringae pv. syrin ...
no activity in no activity in Pseudomonas syringae pv. tomato, Pseudomonas syringae pv. syringae, Pseudomonas syringae pv. syringae B728a
Mol. Plant Pathol.
10
857-865
2009
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
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4
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4
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717051
Sosedov
-
Synthesis of enantiomerically ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Adv. Synth. Catal.
351
1531 - 1538
2009
-
-
1
-
-
-
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5
-
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2
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1
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2
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-
-
1
1
-
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-
684655
Rustler
Simultaneous expression of an ...
Pseudomonas fluorescens, Pseudomonas fluorescens EBC191
Appl. Microbiol. Biotechnol.
80
87-97
2008
-
-
1
-
-
-
-
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-
13
-
-
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-
-
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2
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1
1
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1
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-
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-
-
-
-
-
2
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
687882
Zhu
A new nitrilase from Bradyrhiz ...
Bradyrhizobium japonicum, Bradyrhizobium japonicum USDA 110
J. Biotechnol.
133
327-333
2008
-
-
1
-
-
-
-
1
-
-
3
-
-
24
-
-
1
-
-
-
-
-
26
1
1
-
1
1
1
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
1
-
-
-
-
26
1
1
-
1
1
1
1
-
-
-
-
-
-
-
-
694107
Banerjee
enantioselective nitrilase fro ...
Pseudomonas putida, Pseudomonas putida MTCC 5110
Mol. Biotechnol.
41
35-41
2008
-
1
1
-
-
-
-
-
-
-
1
2
-
2
-
-
1
-
-
-
10
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
2
-
-
-
1
-
-
10
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
700670
Agerbirk
Sinapis phylogeny and evolutio ...
Sinapis alba
Phytochemistry
69
2937-2949
2008
-
-
-
-
-
-
-
-
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1
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3
-
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