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Literature summary extracted from

  • Goda, M.; Hashimoto, Y.; Takase, M.; Herai, S.; Iwahara, Y.; Higashibata, H.; Kobayashi, M.
    Isonitrile hydratase from Pseudomonas putida N19-2. Cloning, sequencing, gene expression, and identification of its active acid residue (2002), J. Biol. Chem., 277, 45860-45865.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.103 gene inhA, DNA and amino acid sequence determination and analysis, overexpression of wild-type and mutant enzyme in Escherichia coli Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.103 C101A site-directed mutagenesis, inactive mutant Pseudomonas putida
4.2.1.103 E79Q site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme Pseudomonas putida
4.2.1.103 E81Q site-directed mutagenesis, slightly decreased activity compared to the wild-type enzyme Pseudomonas putida
4.2.1.103 T102A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Pseudomonas putida

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.103 additional information no metal requirements Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.103 24211
-
 x * 24211, amino acid sequence calculation Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.103 Pseudomonas putida Q8G9F9
-
-
4.2.1.103 Pseudomonas putida N19-2 Q8G9F9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.103 recombinant wild-type and mutants from Escherichia coli Pseudomonas putida

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.1.103 N-cyclohexylformamide = cyclohexyl isocyanide + H2O the enzyme from Pseudomonas putida strain N19-2 can also catalyse the hydration of other isonitriles to the corresponding N-substituted formamides, active site Cys101 and T102 are essential for activity, while Glu79 and Glu81 are not Pseudomonas putida
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.103 1.4
-
 purified recombinant mutant T102A Pseudomonas putida
4.2.1.103 15.4
-
 purified recombinant mutant E81Q Pseudomonas putida
4.2.1.103 17.3
-
 purified recombinant wild-type enzyme Pseudomonas putida
4.2.1.103 18.7
-
 purified recombinant mutant E79Q Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.103 cyclohexyl isocyanide + H2O
-
 Pseudomonas putida N-cyclohexylformamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O
-
 Pseudomonas putida N19-2 N-cyclohexylformamide
-
 ?
4.2.1.103 additional information the enzyme can also catalyse the hydration of other isonitriles to the corresponding N-substituted formamides Pseudomonas putida ?
-
 ?
4.2.1.103 additional information the enzyme can also catalyse the hydration of other isonitriles to the corresponding N-substituted formamides Pseudomonas putida N19-2 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.103 ? x * 24211, amino acid sequence calculation Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
4.2.1.103 isonitrile hydratase
-
 Pseudomonas putida