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Literature summary extracted from

  • Sato, H.; Hashimoto, Y.; Fukatsu, H.; Kobayashi, M.
    Novel isonitrile hydratase involved in isonitrile metabolism (2010), J. Biol. Chem., 285, 34793-34802.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.103 InhB, DNA and amino acid sequence determination and analysis, cloning and expression in Escherichia coli strain DH10B Arthrobacter pascens

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.103 4-chloromercuribenzoate 99.8% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 5,5'-dithiobis-(2-nitrobenzoate) 34.9% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 8-hydroxyquinoline 26.1% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Ag+ complete inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Al3+ 21.4% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Cd2+ 71.6% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Co2+ 89.5% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Cu2+ complete inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Fe2+ 7.6% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Fe3+ 19.7% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 H2O2 45.0% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Hg2+ complete inhibition at 1 mM Arthrobacter pascens
4.2.1.103 additional information the hydrophilic thiol reagent iodoacetate does not influence the enzyme activity. No inhibition by LiCl, NaCl, MgCl, CaCl2, MnCl2, RbCl, CsCl, SrCl2, hydroxylamine, phenylhydrazine, semicarbazide, aminoguanidine, 2,2'-dipyridyl, o-phenanthroline, EDTA, diethyldithiocarbamate, KCN, dithiothreitol, 2-mercaptoethanol, Na2S2O4, and diisopropyl fluorophosphate, poor inhibition by Na2MoO4, ammonium persulfate, and NaN3 Arthrobacter pascens
4.2.1.103 N-ethylmaleimide 48.1% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Ni2+ 95.0% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Pb2+ 11.9% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 PMSF 49.4% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Sn2+ 41.7% inhibition at 1 mM Arthrobacter pascens
4.2.1.103 Zn2+ 51.9% inhibition at 1 mM Arthrobacter pascens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.103 additional information
-
 additional information Michaelis-Menten-type kinetics Arthrobacter pascens
4.2.1.103 0.95
-
 cyclohexyl isocyanide pH 7.5, 25°C Arthrobacter pascens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.103 43000
-
 12 * 43000, SDS-PAGE Arthrobacter pascens
4.2.1.103 530000
-
 gel filtration Arthrobacter pascens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.103 cyclohexyl isocyanide + H2O Arthrobacter pascens
-
 N-cyclohexyl formamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O Arthrobacter pascens F164
-
 N-cyclohexyl formamide
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.103 Arthrobacter pascens E1CJF8
-
-
4.2.1.103 Arthrobacter pascens F164 E1CJF8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.103 native enzyme 14.6fold by ammonium sulfate fractionation followed by two steps of hydrophobic interaction chromatography Arthrobacter pascens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.1.103 cell culture the enzyme is expressed in cells growing on isonitrile as the sole carbon and nitrogen sources, highest enzyme activity is strain F164 in a nutrient medium containing N-benzylformamide. No induction by growth on glycerol and (NH4)2SO4 as the sole carbon and nitrogen sources, respectively Arthrobacter pascens
-

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 0.935
-
 purified enzyme, substrate is cyclohexyl isocyanide, pH 7.5, 25°C Arthrobacter pascens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.103 1-pentyl isocyanide + H2O
-
 Arthrobacter pascens N-1-pentylformamide
-
 ?
4.2.1.103 1-pentyl isocyanide + H2O
-
 Arthrobacter pascens F164 N-1-pentylformamide
-
 ?
4.2.1.103 benzyl isocyanide + H2O
-
 Arthrobacter pascens N-benzylformamide
-
 ?
4.2.1.103 benzyl isocyanide + H2O
-
 Arthrobacter pascens F164 N-benzylformamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O
-
 Arthrobacter pascens N-cyclohexyl formamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O the substrate is partly decomposed into N-cyclohexylformamide on spontaneous and chemical, nonenzymatic reaction under acidic conditions under pH 6.0, but is over pH 6.5 Arthrobacter pascens N-cyclohexyl formamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O
-
 Arthrobacter pascens F164 N-cyclohexyl formamide
-
 ?
4.2.1.103 cyclohexyl isocyanide + H2O the substrate is partly decomposed into N-cyclohexylformamide on spontaneous and chemical, nonenzymatic reaction under acidic conditions under pH 6.0, but is over pH 6.5 Arthrobacter pascens F164 N-cyclohexyl formamide
-
 ?
4.2.1.103 ethyl isocyanoacetate + H2O
-
 Arthrobacter pascens N-formylglycine ethyl ester
-
 ?
4.2.1.103 methyl isocyanoacetate + H2O
-
 Arthrobacter pascens N-formylglycine methyl ester
-
 ?
4.2.1.103 additional information substrate specificity, isocyanomethyl phosphonic acid diethyl ether is a poor substrate giving isocyanomethyl phosphonic acid diethyl ether, overview Arthrobacter pascens ?
-
 ?
4.2.1.103 additional information substrate specificity, isocyanomethyl phosphonic acid diethyl ether is a poor substrate giving isocyanomethyl phosphonic acid diethyl ether, overview Arthrobacter pascens F164 ?
-
 ?
4.2.1.103 tert-butyl isocyanide + H2O
-
 Arthrobacter pascens N-tert-butylformamide
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.103 dodecamer 12 * 43000, SDS-PAGE Arthrobacter pascens

Synonyms

EC Number Synonyms Comment Organism
4.2.1.103 InhB
-
 Arthrobacter pascens
4.2.1.103 isonitrile hydratase
-
 Arthrobacter pascens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.103 25
-
 assay at Arthrobacter pascens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.1.103 35 40 most stable at 35°C, enzyme activity is rapidly lost above 40°C Arthrobacter pascens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.103 7.5
-
 assay at Arthrobacter pascens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
4.2.1.103 5.5 9.7 most stable in the range of pH 5.5-9.7 Arthrobacter pascens

Expression

EC Number Organism Comment Expression
4.2.1.103 Arthrobacter pascens the enzyme is induced by N-benzylformamide and isonitriles benzyl isocyanide and cyclohexyl isocyanide, while benzylamine and N-benzylacetamide do not induce isonitrile hydratase activity up

General Information

EC Number General Information Comment Organism
4.2.1.103 metabolism isonitrile hydratase is involved in the first step of isonitrile metabolism hydrating isonitrile to the corresponding N-substituted formamide Arthrobacter pascens