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Literature summary extracted from

  • Petrickova, A.; Vesela, A.B.; Kaplan, O.; Kubac, D.; Uhnakova, B.; Malandra, A.; Felsberg, J.; Rinagelova, A.; Weyrauch, P.; K?en, V.; Bezouska, K.; Martinkova, L.
    Purification and characterization of heterologously expressed nitrilases from filamentous fungi (2012), Appl. Microbiol. Biotechnol., 93, 1553-1561.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.1 expression in Escherichia coli strain BL21-Gold (DE3) Fusarium verticillioides
3.5.5.1 expression in Escherichia coli strain BL21-Gold (DE3), co-expression of GroEL/ES Talaromyces marneffei
3.5.5.5 expression in Escherichia coli strain BL21-Gold (DE3) Neurospora crassa
3.5.5.5 expression in Escherichia coli strain BL21-Gold (DE3) Aspergillus niger

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.1 0.41
-
 Benzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 0.75
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 0.93
-
 3-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 1.3
-
 4-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 2.2
-
 4-cyanopyridine pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 2.6
-
 4-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 3.2
-
 3-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
3.5.5.1 7.9
-
 Benzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 9.1
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 11.1
-
 3-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 11.7
-
 2-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 15.4
-
 4-cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.1 34.4
-
 3-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.5.5.5 0.8
-
 phenylpropionitrile pH 8.0, 30°C, recombinant enzyme Aspergillus niger
3.5.5.5 1.3
-
 phenylpropionitrile pH 8.0, 30°C, recombinant enzyme Neurospora crassa
3.5.5.5 2
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Neurospora crassa
3.5.5.5 3.4
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Aspergillus niger
3.5.5.5 3.4
-
 2-hydroxy-2-phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Neurospora crassa
3.5.5.5 11.4
-
 2-hydroxy-2-phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Aspergillus niger

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.5.1 2-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 2-carboxylic acid + NH3
-
 ?
3.5.5.1 2-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 2-carboxylic acid + NH3
-
 ?
3.5.5.1 3-chlorobenzonitrile + 2 H2O Talaromyces marneffei
-
 3-chlorobenzoate + NH3
-
 ?
3.5.5.1 3-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 3-carboxylic acid + NH3
-
 ?
3.5.5.1 3-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 3-carboxylic acid + NH3
-
 ?
3.5.5.1 4-chlorobenzonitrile + 2 H2O Talaromyces marneffei
-
 4-chlorobenzoate + NH3
-
 ?
3.5.5.1 4-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 4-carboxylic acid + NH3
-
 ?
3.5.5.1 4-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 4-carboxylic acid + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O Talaromyces marneffei
-
 benzoate + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O Fusarium verticillioides preferred substrate benzoate + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O Talaromyces marneffei ATCC 18224
-
 benzoate + NH3
-
 ?
3.5.5.1 phenylacetonitrile + 2 H2O Talaromyces marneffei
-
 phenylacetate + NH3
-
 ?
3.5.5.1 phenylacetonitrile + 2 H2O Talaromyces marneffei ATCC 18224
-
 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O Neurospora crassa
-
 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O Aspergillus niger
-
 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O Aspergillus niger CBS 513.88
-
 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O Neurospora crassa OR74A
-
 phenylacetate + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.1 Fusarium verticillioides Q19A54
-
-
3.5.5.1 Talaromyces marneffei B6Q5I3
-
-
3.5.5.1 Talaromyces marneffei ATCC 18224 B6Q5I3
-
-
3.5.5.5 Aspergillus niger A2R6M7
-
-
3.5.5.5 Aspergillus niger A9QXE0
-
-
3.5.5.5 Aspergillus niger CBS 513.88 A2R6M7
-
-
3.5.5.5 Aspergillus niger CBS 513.88 A9QXE0
-
-
3.5.5.5 Neurospora crassa
-
-
-
3.5.5.5 Neurospora crassa OR74A
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.5.1 recombinant enzyme 4fold from Escherichia coli strain BL21-Gold (DE3), co-purifiication with GroEL Talaromyces marneffei
3.5.5.1 recombinant enzyme 9fold from Escherichia coli strain BL21-Gold (DE3) co-purifiication with GroEL, the two proteins are further separated by gel filtration Fusarium verticillioides
3.5.5.5 recombinant enzyme 2.4fold from Escherichia coli strain BL21-Gold(DE3) to near homogeneity Neurospora crassa
3.5.5.5 recombinant enzyme 3fold from Escherichia coli strain BL21-Gold(DE3) to near homogeneity Aspergillus niger

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 0.24
-
 recombinant enzyme expressed without GroEL/ES, pH 8.0, 30°C Talaromyces marneffei
3.5.5.1 0.28
-
 recombinant enzyme expressed without GroEL/ES, pH 8.0, 30°C Fusarium verticillioides
3.5.5.1 0.64
-
 recombinant enzyme coexpressed with GroEL/ES, pH 8.0, 30°C Fusarium verticillioides
3.5.5.1 0.7
-
 recombinant enzyme coexpressed with GroEL/ES, pH 8.0, 30°C Talaromyces marneffei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.1 2-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 2-carboxylic acid + NH3
-
 ?
3.5.5.1 2-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 2-carboxylic acid + NH3
-
 ?
3.5.5.1 3-chlorobenzonitrile + 2 H2O
-
 Fusarium verticillioides 3-chlorobenzoate + NH3
-
 ?
3.5.5.1 3-chlorobenzonitrile + 2 H2O
-
 Talaromyces marneffei 3-chlorobenzoate + NH3
-
 ?
3.5.5.1 3-cyanopyridine + 2 H2O
-
 Fusarium verticillioides pyridine 3-carboxylic acid + NH3
-
 ?
3.5.5.1 3-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 3-carboxylic acid + NH3
-
 ?
3.5.5.1 3-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 3-carboxylic acid + NH3
-
 ?
3.5.5.1 4-chlorobenzonitrile + 2 H2O
-
 Fusarium verticillioides 4-chlorobenzoate + NH3
-
 ?
3.5.5.1 4-chlorobenzonitrile + 2 H2O
-
 Talaromyces marneffei 4-chlorobenzoate + NH3
-
 ?
3.5.5.1 4-cyanopyridine + 2 H2O
-
 Fusarium verticillioides pyridine 4-carboxylic acid + NH3
-
 ?
3.5.5.1 4-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 4-carboxylic acid + NH3
-
 ?
3.5.5.1 4-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 4-carboxylic acid + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O
-
 Talaromyces marneffei benzoate + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O preferred substrate Fusarium verticillioides benzoate + NH3
-
 ?
3.5.5.1 benzonitrile + 2 H2O
-
 Talaromyces marneffei ATCC 18224 benzoate + NH3
-
 ?
3.5.5.1 additional information no activity with 2-cyanopyridine Fusarium verticillioides ?
-
 ?
3.5.5.1 phenylacetonitrile + 2 H2O
-
 Fusarium verticillioides phenylacetate + NH3
-
 ?
3.5.5.1 phenylacetonitrile + 2 H2O
-
 Talaromyces marneffei phenylacetate + NH3
-
 ?
3.5.5.1 phenylacetonitrile + 2 H2O
-
 Talaromyces marneffei ATCC 18224 phenylacetate + NH3
-
 ?
3.5.5.5 2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile Neurospora crassa 2-hydroxy-2-phenylacetate + NH3
-
 ?
3.5.5.5 2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile Aspergillus niger 2-hydroxy-2-phenylacetate + NH3
-
 ?
3.5.5.5 2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile Aspergillus niger CBS 513.88 2-hydroxy-2-phenylacetate + NH3
-
 ?
3.5.5.5 2-hydroxy-2-phenylacetonitrile + 2 H2O i.e. (R,S)-mandelonitrile Neurospora crassa OR74A 2-hydroxy-2-phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Neurospora crassa phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Aspergillus niger phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O preferred substrate Neurospora crassa phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O preferred substrate Aspergillus niger phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Aspergillus niger CBS 513.88 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O preferred substrate Aspergillus niger CBS 513.88 phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O
-
 Neurospora crassa OR74A phenylacetate + NH3
-
 ?
3.5.5.5 phenylacetonitrile + 2 H2O preferred substrate Neurospora crassa OR74A phenylacetate + NH3
-
 ?
3.5.5.5 phenylpropionitrile + 2 H2O
-
 Neurospora crassa phenylpropionate + NH3
-
 ?
3.5.5.5 phenylpropionitrile + 2 H2O
-
 Aspergillus niger phenylpropionate + NH3
-
 ?
3.5.5.5 phenylpropionitrile + 2 H2O
-
 Aspergillus niger CBS 513.88 phenylpropionate + NH3
-
 ?
3.5.5.5 phenylpropionitrile + 2 H2O
-
 Neurospora crassa OR74A phenylpropionate + NH3
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.5.1 30
-
 assay at Fusarium verticillioides
3.5.5.1 30
-
 assay at Talaromyces marneffei
3.5.5.5 38
-
-
 Neurospora crassa
3.5.5.5 38
-
-
 Aspergillus niger

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.5.1 25 40 purified recombinant enzyme, pH 8.0, 1 h, stable Fusarium verticillioides
3.5.5.1 25 35 recombinant enzyme in cell-free extract, pH 8.0, 1 h, stable Talaromyces marneffei
3.5.5.1 40
-
 recombinant enzyme in cell-free extract, pH 8.0, 1 h, 20% activity remaining Talaromyces marneffei
3.5.5.1 45
-
 purified recombinant enzyme, pH 8.0, 1 h, 90% activity remaining Fusarium verticillioides
3.5.5.1 50
-
 purified recombinant enzyme, pH 8.0, 1 h, 50% activity remaining Fusarium verticillioides
3.5.5.1 55
-
 and above, purified recombinant enzyme, pH 8.0, 1 h, no activity remaining Fusarium verticillioides
3.5.5.1 55
-
 and above, recombinant enzyme in cell-free extract, pH 8.0, 1 h, no activity remaining Talaromyces marneffei
3.5.5.5 25
-
 purified recombinant enzyme, pH 8.0, 1 h, 90% activity remaining Neurospora crassa
3.5.5.5 25 35 purified recombinant enzyme, pH 8.0, 1 h, stable Aspergillus niger
3.5.5.5 35
-
 purified recombinant enzyme, pH 8.0, 1 h, 80% activity remaining Neurospora crassa
3.5.5.5 40
-
 purified recombinant enzyme, pH 8.0, 1 h, 40% activity remaining Neurospora crassa
3.5.5.5 40
-
 purified recombinant enzyme, pH 8.0, 1 h, 60% activity remaining Aspergillus niger
3.5.5.5 45
-
 and above, purified recombinant enzyme, pH 8.0, 1 h, no activity remaining Neurospora crassa
3.5.5.5 45
-
 purified recombinant enzyme, pH 8.0, 1 h, 20% activity remaining Aspergillus niger
3.5.5.5 50 60 purified recombinant enzyme, pH 8.0, 1 h, about 10% activity remaining Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.5.1 8
-
 assay at Fusarium verticillioides
3.5.5.1 8
-
 assay at Talaromyces marneffei
3.5.5.5 7 9.5
-
 Aspergillus niger
3.5.5.5 8 8.5
-
 Neurospora crassa

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.5.5.1 additional information
-
 sharp drop in activity between pH 11.0 and pH 11.5, profile overview Fusarium verticillioides
3.5.5.1 5.5
-
 purified recombinant enzyme, room temperature, 2 h, 60% activity remaining Fusarium verticillioides
3.5.5.1 6
-
 purified recombinant enzyme, room temperature, 2 h, 80% activity remaining Fusarium verticillioides
3.5.5.1 6.5 11 purified recombinant enzyme, room temperature, 2 h, 90-100% activity remaining Fusarium verticillioides
3.5.5.1 6.5 8.7 recombinant enzyme in cell-free extract, room temperature, 2 h, 70% activity remaining at pH 6.5 and pH 8.7, profile overview Talaromyces marneffei
3.5.5.1 7
-
 purified recombinant enzyme, room temperature, 2 h, completely stable Talaromyces marneffei
3.5.5.1 9.5
-
 and above, purified recombinant enzyme, room temperature, 2 h, no activity remaining Talaromyces marneffei
3.5.5.1 11.5
-
 purified recombinant enzyme, room temperature, 2 h, no activity remaining Fusarium verticillioides
3.5.5.5 5.5
-
 purified recombinant enzyme, room temperature, 2 h, 10% activity remaining Aspergillus niger
3.5.5.5 6.5 9 purified recombinant enzyme, room temperature, 2 h, 80-100% activity remaining Neurospora crassa
3.5.5.5 6.5
-
 purified recombinant enzyme, room temperature, 2 h, 50% activity remaining Aspergillus niger
3.5.5.5 7 10 purified recombinant enzyme, room temperature, 2 h, 90-100% activity remaining Aspergillus niger
3.5.5.5 9.5
-
 purified recombinant enzyme, room temperature, 2 h, 65% activity remaining Neurospora crassa
3.5.5.5 10
-
 purified recombinant enzyme, room temperature, 2 h, 20% activity remaining Neurospora crassa
3.5.5.5 10.5
-
 purified recombinant enzyme, room temperature, 2 h, 20% activity remaining Aspergillus niger
3.5.5.5 11
-
 purified recombinant enzyme, room temperature, 2 h, 10% activity remaining Neurospora crassa
3.5.5.5 11
-
 purified recombinant enzyme, room temperature, 2 h, 50% activity remaining Aspergillus niger

General Information

EC Number General Information Comment Organism
3.5.5.5 evolution the nitrilase from Neurospora crassa is related to aliphatic nitrilase, EC 3.5.5.7 Neurospora crassa