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Literature summary for 3.5.5.1 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
Search for more literature for 3.5.5.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli strain BL21-Gold (DE3) Fusarium verticillioides
expression in Escherichia coli strain BL21-Gold (DE3), co-expression of GroEL/ES Talaromyces marneffei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.41
-
 Benzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
0.75
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
0.93
-
 3-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
1.3
-
 4-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
2.2
-
 4-cyanopyridine pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
2.6
-
 4-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
3.2
-
 3-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Fusarium verticillioides
7.9
-
 Benzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
9.1
-
 phenylacetonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
11.1
-
 3-chlorobenzonitrile pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
11.7
-
 2-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
15.4
-
 4-cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei
34.4
-
 3-Cyanopyridine pH 8.0, 30°C, recombinant enzyme Talaromyces marneffei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 2-carboxylic acid + NH3
-
 ?
2-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 2-carboxylic acid + NH3
-
 ?
3-chlorobenzonitrile + 2 H2O Talaromyces marneffei
-
 3-chlorobenzoate + NH3
-
 ?
3-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 3-carboxylic acid + NH3
-
 ?
3-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 3-carboxylic acid + NH3
-
 ?
4-chlorobenzonitrile + 2 H2O Talaromyces marneffei
-
 4-chlorobenzoate + NH3
-
 ?
4-cyanopyridine + 2 H2O Talaromyces marneffei
-
 pyridine 4-carboxylic acid + NH3
-
 ?
4-cyanopyridine + 2 H2O Talaromyces marneffei ATCC 18224
-
 pyridine 4-carboxylic acid + NH3
-
 ?
benzonitrile + 2 H2O Talaromyces marneffei
-
 benzoate + NH3
-
 ?
benzonitrile + 2 H2O Fusarium verticillioides preferred substrate benzoate + NH3
-
 ?
benzonitrile + 2 H2O Talaromyces marneffei ATCC 18224
-
 benzoate + NH3
-
 ?
phenylacetonitrile + 2 H2O Talaromyces marneffei
-
 phenylacetate + NH3
-
 ?
phenylacetonitrile + 2 H2O Talaromyces marneffei ATCC 18224
-
 phenylacetate + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Fusarium verticillioides Q19A54
-
-
Talaromyces marneffei B6Q5I3
-
-
Talaromyces marneffei ATCC 18224 B6Q5I3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme 4fold from Escherichia coli strain BL21-Gold (DE3), co-purifiication with GroEL Talaromyces marneffei
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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.24
-
 recombinant enzyme expressed without GroEL/ES, pH 8.0, 30°C Talaromyces marneffei
0.28
-
 recombinant enzyme expressed without GroEL/ES, pH 8.0, 30°C Fusarium verticillioides
0.64
-
 recombinant enzyme coexpressed with GroEL/ES, pH 8.0, 30°C Fusarium verticillioides
0.7
-
 recombinant enzyme coexpressed with GroEL/ES, pH 8.0, 30°C Talaromyces marneffei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 2-carboxylic acid + NH3
-
 ?
2-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 2-carboxylic acid + NH3
-
 ?
3-chlorobenzonitrile + 2 H2O
-
 Fusarium verticillioides 3-chlorobenzoate + NH3
-
 ?
3-chlorobenzonitrile + 2 H2O
-
 Talaromyces marneffei 3-chlorobenzoate + NH3
-
 ?
3-cyanopyridine + 2 H2O
-
 Fusarium verticillioides pyridine 3-carboxylic acid + NH3
-
 ?
3-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 3-carboxylic acid + NH3
-
 ?
3-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 3-carboxylic acid + NH3
-
 ?
4-chlorobenzonitrile + 2 H2O
-
 Fusarium verticillioides 4-chlorobenzoate + NH3
-
 ?
4-chlorobenzonitrile + 2 H2O
-
 Talaromyces marneffei 4-chlorobenzoate + NH3
-
 ?
4-cyanopyridine + 2 H2O
-
 Fusarium verticillioides pyridine 4-carboxylic acid + NH3
-
 ?
4-cyanopyridine + 2 H2O
-
 Talaromyces marneffei pyridine 4-carboxylic acid + NH3
-
 ?
4-cyanopyridine + 2 H2O
-
 Talaromyces marneffei ATCC 18224 pyridine 4-carboxylic acid + NH3
-
 ?
benzonitrile + 2 H2O
-
 Talaromyces marneffei benzoate + NH3
-
 ?
benzonitrile + 2 H2O preferred substrate Fusarium verticillioides benzoate + NH3
-
 ?
benzonitrile + 2 H2O
-
 Talaromyces marneffei ATCC 18224 benzoate + NH3
-
 ?
additional information no activity with 2-cyanopyridine Fusarium verticillioides ?
-
 ?
phenylacetonitrile + 2 H2O
-
 Fusarium verticillioides phenylacetate + NH3
-
 ?
phenylacetonitrile + 2 H2O
-
 Talaromyces marneffei phenylacetate + NH3
-
 ?
phenylacetonitrile + 2 H2O
-
 Talaromyces marneffei ATCC 18224 phenylacetate + NH3
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Fusarium verticillioides
30
-
 assay at Talaromyces marneffei

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 40 purified recombinant enzyme, pH 8.0, 1 h, stable Fusarium verticillioides
25 35 recombinant enzyme in cell-free extract, pH 8.0, 1 h, stable Talaromyces marneffei
40
-
 recombinant enzyme in cell-free extract, pH 8.0, 1 h, 20% activity remaining Talaromyces marneffei
45
-
 purified recombinant enzyme, pH 8.0, 1 h, 90% activity remaining Fusarium verticillioides
50
-
 purified recombinant enzyme, pH 8.0, 1 h, 50% activity remaining Fusarium verticillioides
55
-
 and above, purified recombinant enzyme, pH 8.0, 1 h, no activity remaining Fusarium verticillioides
55
-
 and above, recombinant enzyme in cell-free extract, pH 8.0, 1 h, no activity remaining Talaromyces marneffei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Fusarium verticillioides
8
-
 assay at Talaromyces marneffei

pH Stability

pH Stability pH Stability Maximum Comment Organism
additional information
-
 sharp drop in activity between pH 11.0 and pH 11.5, profile overview Fusarium verticillioides
5.5
-
 purified recombinant enzyme, room temperature, 2 h, 60% activity remaining Fusarium verticillioides
6
-
 purified recombinant enzyme, room temperature, 2 h, 80% activity remaining Fusarium verticillioides
6.5 11 purified recombinant enzyme, room temperature, 2 h, 90-100% activity remaining Fusarium verticillioides
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
7
-
 purified recombinant enzyme, room temperature, 2 h, completely stable Talaromyces marneffei
9.5
-
 and above, purified recombinant enzyme, room temperature, 2 h, no activity remaining Talaromyces marneffei
11.5
-
 purified recombinant enzyme, room temperature, 2 h, no activity remaining Fusarium verticillioides