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Literature summary extracted from

  • Pei, X.; Yang, L.; Xu, G.; Wang, Q.; Wu, J.
    Discovery of a new Fe-type nitrile hydratase efficiently hydrating aliphatic and aromatic nitriles by genome mining (2014), J. Mol. Catal. B, 99, 26-33.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.2.1.84 NHase activator protein the activator gene encding activator protein might be involved in protein folding of the alpha- and beta-subunits of NHase Pseudomonas putida

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.84 gene Pput_2728, gene cluster and phylogenetic analysis, functional expression of His6-tagged subunits in Escherichia coli strain BL21(DE3) by assisting of a putative activator gene adjacent to beta-subunit region. Maximal enzyme activity is obtained when the structural and activator genes are transcribed as one unit in plasmid pCDFDuet-1 at 18°C. The expressed product does not show any NHase activity when the downstream activator gene is ignored, and the product completely exists in insoluble inclusion body Pseudomonas putida
4.2.1.84 gene Pput_2729, gene cluster and phylogenetic analysis, functional expression of His6-tagged subunits in Escherichia coli strain BL21(DE3) by assisting of a putative activator gene adjacent to beta-subunit region. Maximal enzyme activity is obtained when the structural and activator genes are transcribed as one unit in plasmid pCDFDuet-1 at 18°C. The expressed product does not show any NHase activity when the downstream activator gene is ignored, and the product completely exists in insoluble inclusion body Pseudomonas putida

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.84 Fe2+ Fe-type NHase Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.84 Pseudomonas putida A5W402 gene Pput_2728 encoding subunit alpha
-
4.2.1.84 Pseudomonas putida A5W402 gene Pput_2729 encoding subunit alpha
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.84 recombinant His6-tagged subunits 2.6fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and desalting gel filtration Pseudomonas putida

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.84 26.61
-
 purified recombinant enzyme, pH 8.0, 25°C, substrate 3-cyanopyridine Pseudomonas putida
4.2.1.84 535
-
 purified recombinant enzyme, pH 8.0, 25°C, substrate valeronitrile Pseudomonas putida
4.2.1.84 553
-
 purified recombinant enzyme, pH 8.0, 25°C, substrate 4-chlorobutyronitrile Pseudomonas putida
4.2.1.84 775
-
 purified recombinant enzyme, pH 8.0, 25°C, substrate isobutyronitrile Pseudomonas putida
4.2.1.84 941
-
 purified recombinant enzyme, pH 8.0, 25°C, substrate acrylonitrile Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.84 3-cyanopyridine + H2O lower activity Pseudomonas putida pyridine-3-carbamide
-
 ?
4.2.1.84 3-methylbenzonitrile + H2O lower activity Pseudomonas putida 3-methylbenzamide
-
 ?
4.2.1.84 4-chlorobutyronitrile + H2O
-
 Pseudomonas putida 4-chlorobutyramide
-
 ?
4.2.1.84 4-cyanopyridine + H2O lower activity Pseudomonas putida isonicotinamide
-
 ?
4.2.1.84 acrylonitrile + H2O best substrate Pseudomonas putida acrylamide
-
 ?
4.2.1.84 benzeneacetonitrile + H2O very low activity Pseudomonas putida ?
-
 ?
4.2.1.84 benzonitrile + H2O lower activity Pseudomonas putida benzamide
-
 ?
4.2.1.84 isobutyronitrile + H2O
-
 Pseudomonas putida isobutyramide
-
 ?
4.2.1.84 additional information the Fe-type NHase exhibits broad substrate specificity Pseudomonas putida ?
-
 ?
4.2.1.84 valeronitrile + H2O
-
 Pseudomonas putida ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.84 NHase
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.84 35
-
-
 Pseudomonas putida

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.2.1.84 10 45 activity range, profile overview Pseudomonas putida

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.1.84 20
-
 purified recombinant enzyme, over 60% activity remaining after 3 h Pseudomonas putida
4.2.1.84 20 45 purified recombinant enzyme, 50% of maximal activity within this range. Half-lives of NHase F1 at 20°C, 25°C, 30°C, and 35°C are calculated as 5.1 h, 3.8 h, 2.2 h, and 1.2 h, respectively Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.84 8
-
-
 Pseudomonas putida

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.84 6 10 activity range, profile overview Pseudomonas putida

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
4.2.1.84 7 8 purified recombinant enzyme, 25°C, highest stability at Pseudomonas putida

General Information

EC Number General Information Comment Organism
4.2.1.84 evolution because of the absence of sequence similarity between the Co- and Fe-type NHase activators, the two types of NHases might be assembled and maturated by different molecular mechanisms Pseudomonas putida