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4.2.1.84: nitrile hydratase

This is an abbreviated version!
For detailed information about nitrile hydratase, go to the full flat file.

Word Map on EC 4.2.1.84

Reaction

an aliphatic amide
=
A nitrile
 +
H2O

Synonyms

3-cyanopyridine hydratase, acrylonitrile hydratase, aliphatic nitrile hydratase, ANHase, Co-type NHase, Co-type nitrile hydratase, cobalt-containing nitrile hydratase, CoIII-NHase, CtNHase, Fe-NHase, H-NHase, H-nitrilase, high-molecular mass nitrile hydratase, high-molecular weight nitrile hydratase, hydratase, nitrile, iron-type nitrile hydratase, L-Nhase, L-nitrilase, low-molecular mass nitrile hydratase, low-molecular weight nitrile hydratase, MbNHase, NHase, NHaseK, NI1 NHase, NilCo, NilFe, nitrilase, nitrile hydratase, NthAB, PaNit, ppNHase, ReNHase, TNHase, toyocamycin nitrile hydratase

ECTree

     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.1 Hydro-lyases
                4.2.1.84 nitrile hydratase

Purification

Purification on EC 4.2.1.84 - nitrile hydratase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
(NH4)2SO4 fractionation
-
(NH4)2SO4 fractionation, ion-exchange/hydrophobic/gel-filtration chromatography, stabilization by n-butyric acid, yield of purified protein: 23.96%
-
3 successive column chromatographies in the presence of 40 mM n-butyric acid (only wild type), alphaQ90N mutein is purified in nitrosylated state in the dark
-
ammonium sulfate fractionation and chromatography on Q-Sepharose, phenyl-Sepharose and Sephadex G150
-
ammonium sulfate fractionation, chromatography on DEAE-Cellulofine, phenyl-Sepharose, Sephadex G-150 and octyl-Sepharose, 11.5fold purification, N-774
-
ammonium sulfate fractionation, chromatography on DEAE-Sephacel and phenyl-Sepharose CL-4B and gel filtration, R312 strain, 10.2fold purified
-
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, octyl and phenyl-Sepharose CL-4B
-
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, octyl-Sepharose CL-4B and phenyl-Sepharose CL-4B, 4.91fold purification
-
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose and Sephacryl S-300, 3.31fold purification
-
ammonium sulfate fractionation, chromatography on DEAE-Sephacel, phenyl-Sepharose CL-4B and gel filtration
-
ammonium sulfate fractionation, Sephacryl S 300 gel filtration, and DEAE column chromatography
-
ammonium sulfate precipitation and Q-Sepharose column chromatography
-
ammonium sulfate precipitation, ion exchange chromatography and chromatography on phenyl-Sepharose CL-4B, 11fold purification, mutant strain ACV2
-
anion exchange and hydrophobic interaction chromatography, yield of purified protein: 1.9% (specific activity 5.9 U/mg), cytosolic proteom of cells grown on 0.1 M acetonitrile and acetic acid/ammonia are compared via quantitative 2D gel electrophoresis, spot identification by MS, no significant amino acid sequence similarity with other NHases
-
anion exchange chromatography and gel filtration, recombinant enzyme
-
cell-free extract preparation
-
chromatography on butyl-toyopearl
-
chromatography on DEAE-Cellulose, DEAE-Sephacel and phenyl-Sepharose CL-4B, 130fold purification
-
chromatography on DEAE-Cellulose, hydroxyapatite, phenyl-Sepharose and Sephacryl S-400
-
chromatography on phenyl-Sepharose CL-4B and Bio-Gel HTP
-
chromatography on Q-Sepharose, phenyl-Sepharose and Uno Q
-
chromatography on Sepharose Q, phenyl-Sepharose CL-4B and Sephadex G-200, strain 7
-
CoCl2, an overexpressed chaperonin system and butyric acid is necessary for fully soluble and active NilCo protein, 95% purity on 12% SDS-PAGE
-
native enzyme 52fold by ultracentrifugation and ammonium sulfate fractionation of the supernatant, followed by ion exchange chromatography and gel filtration
-
native enzyme 78fold by ammonium sulfate fractionation, anion exchange chromatography, and hydrophobic interaction chromatography
-
native enzyme from strain F28 5fold by ammonium sulfate fractionation, anion exchange chromatography and gel filtration
-
native enzyme to homogeneity
-
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant enzyme from Escherichia coli strain HB101 by heat treatment at 55°C for 30 min, ammonium sulfate precipitation, anion exchange chromatography, hydrophobic interaction chromatography, and a another differentstep of anion exchange chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography and ultrafiltration
-
recombinant His-tagged wild-type enzyme and peptide fused enzyme variants from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
recombinant His6-tagged subunits 2.6fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and desalting gel filtration
recombinant L-NHase, NhlAE, and the NhhA-NhlE hybrid mediator complex from Rhodococcus rhodochrous strain ATCC12674 and Rhodococcus fascians DSM43985 by ammonium sulfate fractionation anion exchange chromatography, dialysis, gel filtration, and another step of anion exchange chromatography
-
recombinant Nit from Escherichia coli strain Bl21(DE3) by ammonium sulfate fractionation and anion exchange chromatography
-
recombinant soluble wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography, ammonium sulfate fractionation, and hydrophobic interaction chromatography, followed by ultrafiltration and another step of anion exchange chromatography
-
recombinant soluble wild-type and mutant enzymes from Escherichia coli strain JM109 by ammonium sulfate fractionation and two different steps of anion echange chromatography
Resource Q column chromatography and Superdex-200 gel filtration
-
Sepharose 6 column chromatography
-
sonication, (NH4)2SO4 fractionation, DEAE-Sephacel/phenyl Sepharose/Sephacryl S200 column chromatography
-
the chimeric NHase (SBpNHase) from the thermal sensitive nitrile hydratasese from Bordetella petrii and the relatively thermal-stable nitrile hydratases from Pseudonocardia thermophila is constructed by swapping the corresponding C-domains
to homogeneity, using several chromatographic steps, including DEAE, hydroxyapatite, and Sephadex G-200 column chromatography