Cloned (Comment) | Organism |
---|---|
gene nit-30, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain Bl21(DE3) | Pyrococcus abyssi |
Crystallization (Comment) | Organism |
---|---|
PaNit without ligands, and with an acetate ion bound in the active site, or with a bromide ion in the active site, by hanging drop vapour diffusion method from 35% PEG 550MME, 0.2 M MgCl2 or Mg(CH3COOH)2 and 0.2 M Tris-HCl, pH 7.5, X-ray diffraction structure determination and analysis at 1.57-1.83 A resolution | Pyrococcus abyssi |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
acetate | the acetate ion binds close to Mg2+, and it interacts with two coordinating water molecules | Pyrococcus abyssi | |
Mg2+ | Mg2+ bound in chain B interacts with five water molecules and Asn49B | Pyrococcus abyssi | |
additional information | specific binding of the carboxylate group, as well as a more general electrostatic preference for negatively charged ligands revealed by binding of the Br- ions, overview | Pyrococcus abyssi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Pyrococcus abyssi | the thermoactive nitrilase from Pyrococcus abyssi hydrolyses small aliphatic nitriles like fumaro- and malononitril, docking calculations for fumaro- and malononitriles, modelling, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus abyssi | - |
gene nit-30 | - |
Purification (Comment) | Organism |
---|---|
recombinant Nit from Escherichia coli strain Bl21(DE3) by ammonium sulfate fractionation and anion exchange chromatography | Pyrococcus abyssi |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
an aliphatic amide = a nitrile + H2O | role of residues in the active site and enzymatic reaction mechanism. Cys146 acts as the nucleophile, Glu42 as the general base, Lys113/Glu42 as the general acid, WatA as the hydrolytic water and Nf_Lys113 and N_Phe147 form the oxyanion hole, hydrogen bonding network in the active site of Nit structure, overview | Pyrococcus abyssi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the thermoactive nitrilase from Pyrococcus abyssi hydrolyses small aliphatic nitriles like fumaro- and malononitril, docking calculations for fumaro- and malononitriles, modelling, overview | Pyrococcus abyssi | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
nitrilase | - |
Pyrococcus abyssi |
PaNit | - |
Pyrococcus abyssi |