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Literature summary extracted from

  • Stalker, D.M.; Malyj, L.D.; McBride, K.E.
    Purification and properties of a nitrilase specific for the herbicide bromoxynil and corresponding nucleotide sequence analysis of the bxn gene (1988), J. Biol. Chem., 263, 6310-6314.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.5.5.6 agriculture recent experiments indicate that expression of this bacterial nitrilase in transgenic plants results in high levels of resistance to the herbicide bromoxynil Klebsiella pneumoniae subsp. ozaenae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.5.6 DNA segment cloned and expressed in Escherichia coli MM 294, nucleotide sequence analysis of the bxn gene Klebsiella pneumoniae subsp. ozaenae

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.5.6 Ag+ 0.01 mM, 93.8% of maximum activity, 0.1 mM: 2.5% of maximal activity Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 Cu2+ 0.01 mM, 75% of maximal activity, 0.1 mM: 31.3% of maximal activity Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 iodoacetate 0.01 mM: 5% of maximal activity Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 additional information Ca2+, Co2+, Mg2+, Mn2+, Zn2+ at 0.1 mM, as well as EDTA or HCN, or monovalent cations at 500 mM are no inhibitors Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 p-hydroxymercuribenzoate 0.0005 mM, 2.5% of maximal activity Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 Phenylmercuriacetate 0.0005 mM, 4.3% of maximal activity Klebsiella pneumoniae subsp. ozaenae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.5.6 0.31
-
 3,5-dibromo-4-hydroxybenzonitrile pH 9.2, 35°C Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 0.55
-
 3,5-Diiodo-4-hydroxybenzonitrile pH 9.2, 35°C Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 0.83
-
 3,5-Dichloro-4-hydroxybenzonitrile pH 9.2, 35°C Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 0.91
-
 5-Bromo-4-hydroxybenzonitrile pH 9.2, 35°C Klebsiella pneumoniae subsp. ozaenae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.5.6 37000
-
 2 * 37000, SDS-PAGE Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 72000
-
 recombinant enzyme, gel filtration Klebsiella pneumoniae subsp. ozaenae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.5.6 3,5-dibromo-4-hydroxybenzonitrile + H2O Klebsiella pneumoniae subsp. ozaenae involved in bacterial degradation of the herbizide bromoxynil 3,5-dibromo-4-hydroxybenzoic acid + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.5.6 Klebsiella pneumoniae subsp. ozaenae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.5.6 recombinant enzyme Klebsiella pneumoniae subsp. ozaenae

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.5.6 25.7
-
 3,5-dibromo-4-hydroxybenzonitrile Klebsiella pneumoniae subsp. ozaenae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.5.6 3,5-dibromo-4-hydroxybenzonitrile + H2O bromoxynil Klebsiella pneumoniae subsp. ozaenae 3,5-dibromo-4-hydroxybenzoic acid + NH3
-
 ?
3.5.5.6 3,5-dibromo-4-hydroxybenzonitrile + H2O involved in bacterial degradation of the herbizide bromoxynil Klebsiella pneumoniae subsp. ozaenae 3,5-dibromo-4-hydroxybenzoic acid + NH3
-
 ?
3.5.5.6 3,5-dichloro-4-hydroxybenzonitrile + H2O chloroxynil Klebsiella pneumoniae subsp. ozaenae 3,5-dichloro-4-hydroxybenzoic acid + NH3
-
 ?
3.5.5.6 3,5-diiodo-4-hydroxybenzonitrile + H2O ioxynil Klebsiella pneumoniae subsp. ozaenae 3,5-diiodo-4-hydroxybenzoic acid + NH3
-
 ?
3.5.5.6 5-bromo-4-hydroxybenzonitrile + H2O
-
 Klebsiella pneumoniae subsp. ozaenae 5-bromo-4-hydroxybenzoic acid + NH3
-
 ?
3.5.5.6 additional information no detectable activity when benzonitrile is utilized as substrate, nor does 3,5-dibromo-4-hydroxybenzamide act as a substrate for the enzyme Klebsiella pneumoniae subsp. ozaenae ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.5.6 dimer 2 * 37000, SDS-PAGE Klebsiella pneumoniae subsp. ozaenae
3.5.5.6 dimer 2 * 38100,homodimer, nucleotide sequence analysis Klebsiella pneumoniae subsp. ozaenae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.5.6 35
-
-
 Klebsiella pneumoniae subsp. ozaenae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.5.6 9.2
-
-
 Klebsiella pneumoniae subsp. ozaenae

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.5.6 5.5 10.5 15% of optimal activity demonstrated at pH 7.0, with a rapid rise to pH 9.2 and a steep decline approaching pH 10.6 Klebsiella pneumoniae subsp. ozaenae

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.5.6 Klebsiella pneumoniae subsp. ozaenae calculated pI, derived from the amino acid composition, this is in contrast to the final purification step using a chromatofocusing column, the active form of the enzyme binds to the column at pH 4.0 and must be eluted with high salt concentrations, this observation indicates that the actual pI for the bromoxynil-specific nitrilase must be lower than 4.0
-
 5.4