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Literature summary for 3.5.5.6 extracted from
- 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.
Application
| Application | Comment | Organism |
|---|---|---|
| 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)
| Cloned (Comment) | Organism |
|---|---|
| DNA segment cloned and expressed in Escherichia coli MM 294, nucleotide sequence analysis of the bxn gene | Klebsiella pneumoniae subsp. ozaenae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ag+ | 0.01 mM, 93.8% of maximum activity, 0.1 mM: 2.5% of maximal activity | Klebsiella pneumoniae subsp. ozaenae |  |
| Cu2+ | 0.01 mM, 75% of maximal activity, 0.1 mM: 31.3% of maximal activity | Klebsiella pneumoniae subsp. ozaenae | |
| iodoacetate | 0.01 mM: 5% of maximal activity | Klebsiella pneumoniae subsp. ozaenae | |
| 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 | |
| p-hydroxymercuribenzoate | 0.0005 mM, 2.5% of maximal activity | Klebsiella pneumoniae subsp. ozaenae | |
| Phenylmercuriacetate | 0.0005 mM, 4.3% of maximal activity | Klebsiella pneumoniae subsp. ozaenae | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.31 | - |
3,5-dibromo-4-hydroxybenzonitrile | pH 9.2, 35°C | Klebsiella pneumoniae subsp. ozaenae | |
| 0.55 | - |
3,5-Diiodo-4-hydroxybenzonitrile | pH 9.2, 35°C | Klebsiella pneumoniae subsp. ozaenae | |
| 0.83 | - |
3,5-Dichloro-4-hydroxybenzonitrile | pH 9.2, 35°C | Klebsiella pneumoniae subsp. ozaenae | |
| 0.91 | - |
5-Bromo-4-hydroxybenzonitrile | pH 9.2, 35°C | Klebsiella pneumoniae subsp. ozaenae | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 37000 | - |
2 * 37000, SDS-PAGE | Klebsiella pneumoniae subsp. ozaenae |
| 72000 | - |
recombinant enzyme, gel filtration | Klebsiella pneumoniae subsp. ozaenae |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Klebsiella pneumoniae subsp. ozaenae | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme | Klebsiella pneumoniae subsp. ozaenae |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 25.7 | - |
3,5-dibromo-4-hydroxybenzonitrile | Klebsiella pneumoniae subsp. ozaenae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3,5-dibromo-4-hydroxybenzonitrile + H2O | bromoxynil | Klebsiella pneumoniae subsp. ozaenae | 3,5-dibromo-4-hydroxybenzoic acid + NH3 | - |
? | |
| 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-dichloro-4-hydroxybenzonitrile + H2O | chloroxynil | Klebsiella pneumoniae subsp. ozaenae | 3,5-dichloro-4-hydroxybenzoic acid + NH3 | - |
? | |
| 3,5-diiodo-4-hydroxybenzonitrile + H2O | ioxynil | Klebsiella pneumoniae subsp. ozaenae | 3,5-diiodo-4-hydroxybenzoic acid + NH3 | - |
? | |
| 5-bromo-4-hydroxybenzonitrile + H2O | - |
Klebsiella pneumoniae subsp. ozaenae | 5-bromo-4-hydroxybenzoic acid + NH3 | - |
? | |
| 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
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 37000, SDS-PAGE | Klebsiella pneumoniae subsp. ozaenae |
| dimer | 2 * 38100,homodimer, nucleotide sequence analysis | Klebsiella pneumoniae subsp. ozaenae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
- |
Klebsiella pneumoniae subsp. ozaenae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9.2 | - |
- |
Klebsiella pneumoniae subsp. ozaenae |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 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
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| 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 |
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