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Literature summary for 4.2.1.84 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

Activating Compound Comment Organism Structure
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)

Cloned (Comment) Organism
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
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

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

Organism

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

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
NHase
-
Pseudomonas putida

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
20
-
purified recombinant enzyme, over 60% activity remaining after 3 h Pseudomonas putida
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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Pseudomonas putida

pH Range

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

pH Stability

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

General Information

General Information Comment Organism
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