Literature summary for 4.2.1.103 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.

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Cloned(Commentary)

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

Protein Variants

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

Metals/Ions

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

Molecular Weight [Da]

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

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	Q8G9F9	-	-
Pseudomonas putida N19-2	Q8G9F9	-	-

Purification (Commentary)

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

Reaction

Reaction	Comment	Organism	Reaction ID
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]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
isonitrile hydratase	-	Pseudomonas putida

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Release 2023.1 (January 2023)