The enzyme from Pseudomonas putida strain N19-2 can also catalyse the hydration of other isonitriles to the corresponding N-substituted formamides. The enzyme has no metal requirements.
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
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
The enzyme from Pseudomonas putida strain N19-2 can also catalyse the hydration of other isonitriles to the corresponding N-substituted formamides. The enzyme has no metal requirements.
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
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
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
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
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
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
isocyanide hydratase is an enzyme of the DJ-1 superfamily that hydrates isocyanides to yield the corresponding N-formamide, structural basis for catalysis, overview. ICH contains a highly conserved Cys101 that is required for catalysis and interacts with Asp17, Thr102, and an ordered water molecule in the active site. Asp17 activates the ordered water molecule to hydrate organic isocyanides. The thiolate of Cys101 is proposed to be a critical nucleophile that initiates the hydration of isocyanides
isocyanide hydratase is an enzyme of the DJ-1 superfamily that hydrates isocyanides to yield the corresponding N-formamide, structural basis for catalysis, overview. ICH contains a highly conserved Cys101 that is required for catalysis and interacts with Asp17, Thr102, and an ordered water molecule in the active site. Asp17 activates the ordered water molecule to hydrate organic isocyanides. The thiolate of Cys101 is proposed to be a critical nucleophile that initiates the hydration of isocyanides
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type and several site-directed mutants, hanging drop vapor diffusion method, for wild-type, C101S, and C101A enzymes: 0.002 ml of 20 mg/ml protein solution is mixed with 0.002 ml of reservoir solution containing 24-26% PEG 3350, 200-250 mM magnesium chloride, 100 mM CHES, pH 9.3, or 100 mM Tris-HCl, pH 8.6, and equilibration against 0.5 ml of reservoir solution, 1-3 days, for D17E enzyme: 0.002 ml of 24 mg/ml protein is mixed with 0.002 ml of reservoir solution containing 12% PEG 4000, 240 mM ammonium acetate, 100 mM sodium acetate, pH 4.6, 2-3 days at room temperature, for T102S enzyme: 0.002 ml of 17 mg/ml protein solution is mixed with 0.002 ml of reservoir solution containing 19-22% PEG 4000, 140-160 mM sodium citrate, pH 5.6, 2-3 days at room temperature, X-ray diffraction structure determination and analysis at 1.0-1.9 A resolution
site-directed mutagenesis, inactive mutant, the mutant shows a stronger electron density for a water molecule between Asp17 and residue 101 compared to the wild-type enzyme
site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme, the mutation results in a substrate-inhibited enzyme
site-directed mutagenesis, inactive mutant, the mutant shows a stronger electron density for a water molecule between Asp17 and residue 101 compared to the wild-type enzyme
recombinant N-terminally His-tagged wild-type and mutant ICHs from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the His-tag through thrombin, and benzamidine affinity chromatography. The final recombinant ICH protein has three vector-derived amino acids at the N terminus, GSH
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
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
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
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