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Literature summary for 4.8.1.2 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
Search for more literature for 4.8.1.2

Activating Compound

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

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis-2-nitrobenzoate weak inhibition Pseudomonas chlororaphis
AgNO3 very sensitive to, concentration-dependent, 1 mM, 85.8% inhibition Pseudomonas chlororaphis
CO 49% inhibition Pseudomonas chlororaphis
diethyldithiocarbamate 1 mM, 40.6% inhibition Pseudomonas chlororaphis
hydroxylamine concentration-dependent, 1 mM, 100% inhibition Pseudomonas chlororaphis
iodoacetate weak inhibition Pseudomonas chlororaphis
KCN 1 mM, 41.3% inhibition Pseudomonas chlororaphis
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
N-ethylmaleimide weak inhibition Pseudomonas chlororaphis
p-chloromercuribenzoate weak inhibition Pseudomonas chlororaphis
phenylhydrazine 1 mM, 71.1% inhibition Pseudomonas chlororaphis

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ contains 1.58 mol Ca2+ per mol of homodimeric OxdA, may act as another cofactor Pseudomonas chlororaphis
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Pseudomonas chlororaphis
-
 B23
-
Pseudomonas chlororaphis Q7WSJ4
-
-
Pseudomonas chlororaphis B23
-
 B23
-
Pseudomonas chlororaphis B23 Q7WSJ4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant OxdA Pseudomonas chlororaphis

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
aldoxime dehydratase
-
 Pseudomonas chlororaphis
OxdA
-
 Pseudomonas chlororaphis

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

pH Range

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

Cofactor

Cofactor Comment Organism Structure
additional information no requirement of FMN Pseudomonas chlororaphis
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