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Literature summary extracted from

  • Oinuma, K-I.; Hashimoto, Y.; Konishi, K.; Goda, M.; Noguchi, T.; Higashibata, H.; Kobayashi, M.
    Novel aldoxime dehydratase involved in carbon-nitrogen triple bond synthesis of Pseudomonas chlororaphis B23: Sequencing, gene expression, purification and characterization (2003), J. Biol. Chem., 278, 29600-29608.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.8.1.2 Na2S2O4 requirement of a reducing agent for activity Pseudomonas chlororaphis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.8.1.2 oxdA gene, overexpression in Escherichia coli BL21-Codon-Plus(DE3)-RIL, sequencing, gene cluster organisation Pseudomonas chlororaphis

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.8.1.2 5,5'-dithiobis-2-nitrobenzoate weak inhibition Pseudomonas chlororaphis
4.8.1.2 AgNO3 very sensitive to, concentration-dependent, 1 mM, 85.8% inhibition Pseudomonas chlororaphis
4.8.1.2 CO 49% inhibition Pseudomonas chlororaphis
4.8.1.2 diethyldithiocarbamate 1 mM, 40.6% inhibition Pseudomonas chlororaphis
4.8.1.2 hydroxylamine concentration-dependent, 1 mM, 100% inhibition Pseudomonas chlororaphis
4.8.1.2 iodoacetate weak inhibition Pseudomonas chlororaphis
4.8.1.2 KCN 1 mM, 41.3% inhibition Pseudomonas chlororaphis
4.8.1.2 additional information not inhibited by chelating agents, such as 2,2’-dipyridyl, o-phenanthroline, 8-hydroxyquinoline, EDTA, and by phenylmethanesulfonyl fluoride, diisopropyl fluorophosphate Pseudomonas chlororaphis
4.8.1.2 N-ethylmaleimide weak inhibition Pseudomonas chlororaphis
4.8.1.2 p-chloromercuribenzoate weak inhibition Pseudomonas chlororaphis
4.8.1.2 phenylhydrazine 1 mM, 71.1% inhibition Pseudomonas chlororaphis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.8.1.2 0.25
-
 (E/Z)-n-butyraldoxime pH 7, 30°C Pseudomonas chlororaphis
4.8.1.2 3.4
-
 (E/Z)-pyridine-2-aldoxime pH 7, 30°C Pseudomonas chlororaphis
4.8.1.2 11
-
 (E/Z)-acetaldoxime pH 7, 30°C Pseudomonas chlororaphis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.8.1.2 Ca2+ contains 1.58 mol Ca2+ per mol of homodimeric OxdA, may act as another cofactor Pseudomonas chlororaphis
4.8.1.2 Fe2+ contains 1.62 mol iron per mol of homodimeric OxdA, present in the heme molecule, the heme iron of the active enzyme is in the ferrous state Pseudomonas chlororaphis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.8.1.2 38000
-
 2 * 40127, calculated from the amino acid sequence, 2 * 38000, SDS-PAGE Pseudomonas chlororaphis
4.8.1.2 40127
-
 2 * 40127, calculated from the amino acid sequence, 2 * 38000, SDS-PAGE Pseudomonas chlororaphis
4.8.1.2 76200
-
 OxdA reduced by Na2S2O4, gel filtration Pseudomonas chlororaphis
4.8.1.2 76400
-
 OxdA without reducing conditions, gel filtration Pseudomonas chlororaphis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.8.1.2 additional information Pseudomonas chlororaphis involved in carbon-nitrogen triple bond synthesis, responsible for the metabolism of aldoxime in vivo ?
-
 ?
4.8.1.2 additional information Pseudomonas chlororaphis B23 involved in carbon-nitrogen triple bond synthesis, responsible for the metabolism of aldoxime in vivo ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.8.1.2 Pseudomonas chlororaphis
-
 B23
-
4.8.1.2 Pseudomonas chlororaphis Q7WSJ4
-
-
4.8.1.2 Pseudomonas chlororaphis B23
-
 B23
-
4.8.1.2 Pseudomonas chlororaphis B23 Q7WSJ4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.8.1.2 recombinant OxdA Pseudomonas chlororaphis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.8.1.2 0.758
-
 pH 7, 30°C, butyraldoxime as substrate, under aerobic conditions Pseudomonas chlororaphis
4.8.1.2 197
-
 pH 7, 30°C, butyraldoxime as substrate, under anaerobic reduced conditions, OxdA reduced by Na2S2O4 Pseudomonas chlororaphis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.8.1.2 (E/Z)-acetaldoxime
-
 Pseudomonas chlororaphis acetonitrile + H2O
-
 ?
4.8.1.2 (E/Z)-acetaldoxime
-
 Pseudomonas chlororaphis B23 acetonitrile + H2O
-
 ?
4.8.1.2 (E/Z)-n-butyraldoxime best substrate, accepts both the E- and the Z-forms of butyraldoxime as substrates Pseudomonas chlororaphis n-butyronitrile + H2O butyronitrile does not act as substrate ?
4.8.1.2 (E/Z)-n-butyraldoxime best substrate, accepts both the E- and the Z-forms of butyraldoxime as substrates Pseudomonas chlororaphis B23 n-butyronitrile + H2O butyronitrile does not act as substrate ?
4.8.1.2 (E/Z)-pyridine-2-aldoxime poor substrate Pseudomonas chlororaphis pyridine-2-nitrile + H2O
-
 ?
4.8.1.2 aliphatic aldoxime strong preference of aliphatic aldoximes over aromatic aldoximes Pseudomonas chlororaphis aliphatic nitrile + H2O
-
 ?
4.8.1.2 aliphatic aldoxime strong preference of aliphatic aldoximes over aromatic aldoximes Pseudomonas chlororaphis B23 aliphatic nitrile + H2O
-
 ?
4.8.1.2 an aliphatic aldoxime
-
 Pseudomonas chlororaphis an aliphatic nitrile + H2O
-
 ?
4.8.1.2 an aliphatic aldoxime
-
 Pseudomonas chlororaphis B23 an aliphatic nitrile + H2O
-
 ?
4.8.1.2 additional information not: butyronitrile, aromatic aldoximes, such as benzaldoxime, isonitrosoacetophenone, and pyridine-4-aldoxime Pseudomonas chlororaphis ?
-
 ?
4.8.1.2 additional information involved in carbon-nitrogen triple bond synthesis, responsible for the metabolism of aldoxime in vivo Pseudomonas chlororaphis ?
-
 ?
4.8.1.2 additional information not: butyronitrile, aromatic aldoximes, such as benzaldoxime, isonitrosoacetophenone, and pyridine-4-aldoxime Pseudomonas chlororaphis B23 ?
-
 ?
4.8.1.2 additional information involved in carbon-nitrogen triple bond synthesis, responsible for the metabolism of aldoxime in vivo Pseudomonas chlororaphis B23 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.8.1.2 homodimer 2 * 40127, calculated from the amino acid sequence, 2 * 38000, SDS-PAGE Pseudomonas chlororaphis

Synonyms

EC Number Synonyms Comment Organism
4.8.1.2 aldoxime dehydratase
-
 Pseudomonas chlororaphis
4.8.1.2 OxdA
-
 Pseudomonas chlororaphis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.8.1.2 45
-
-
 Pseudomonas chlororaphis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.8.1.2 20
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 100% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 25
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 94% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 30
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 91% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 35
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 78% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 40
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 51% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 45
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 14% of enzyme activity Pseudomonas chlororaphis
4.8.1.2 50
-
 after 15 min preincubation OxdA is reduced by Na2S2O4, under anaerobic conditions, 3.2% of enzyme activity Pseudomonas chlororaphis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.8.1.2 5.4
-
 (E/Z)-pyridine-2-aldoxime pH 7, 30°C Pseudomonas chlororaphis
4.8.1.2 324
-
 (E/Z)-n-butyraldoxime pH 7, 30°C Pseudomonas chlororaphis
4.8.1.2 336
-
 (E/Z)-acetaldoxime pH 7, 30°C Pseudomonas chlororaphis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.8.1.2 additional information
-
 maximum at pH 5.5, another high activity level occurs at pH 9.4 Pseudomonas chlororaphis
4.8.1.2 5.5
-
-
 Pseudomonas chlororaphis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.8.1.2 5 10 active over a broad pH-range Pseudomonas chlororaphis

Cofactor

EC Number Cofactor Comment Organism Structure
4.8.1.2 additional information no requirement of FMN Pseudomonas chlororaphis
4.8.1.2 protoheme IX hemoprotein, the heme iron of the active enzyme is in the ferrous state, carries protoheme IX as the prosthetic group, 0.69 mol heme per mol of subunit Pseudomonas chlororaphis