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Literature summary for 3.5.1.66 extracted from

  • Huynh, M.S.; Snell, E.E.
    Enzymes of vitamin B6 degradation. Purification and properties of two N-acetylamidohydrolases (1985), J. Biol. Chem., 260, 2379-2383.
    View publication on PubMed
Search for more literature for 3.5.1.66

Inhibitors

Inhibitors Comment Organism Structure
Dicarboxylic acids competitive Arthrobacter sp.
diphosphate competitive Arthrobacter sp.
Hg2+ 0.01 mM, 15 min, 82% activity remains Arthrobacter sp.
Itaconic acid competitive, most effective inhibitor, Ki: 0.74 mM Arthrobacter sp.
malonate competitive, Ki: 20 mM Arthrobacter sp.
additional information no inhibition by: organic protein reagents, 0.1 mM, 15 min, p-chloromercuribenzoate, 1.0 mM Ni2+, 1.0 mM Co2+, 1.0 mM Mn2+, 1.0 mM Mg2+, 1.0 mM Ca2+, 1.0 mM Cu2+, 1.0 mM Cs+, 1.0 mM Li+, SO42-, NO3-, Cl-, 10 mM dithiothreitol, 100 mM monocarboxylic acids Arthrobacter sp.
phosphate competitive Arthrobacter sp.
sulfite noncompetitive, inhibition by a time-dependent mechanism, addition of succinic semialdehyde protects against sulfite inhibition Arthrobacter sp.
Tricarboxylic acids competitive Arthrobacter sp.
Zn2+ 1 mM: 40% inhibition Arthrobacter sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic data Arthrobacter sp.
0.025
-
 2-(Hydroxymethyl)-3-(acetamidomethylene)succinate compound B, 25°C, Tris-HCl, pH 7.0 Arthrobacter sp.
0.123
-
 2-(Hydroxymethyl)-3-(acetamidomethylene)succinate compound B, 2-(N-morpholino)ethane-sulfonic acid, pH 6.0 Arthrobacter sp.
0.25
-
 2-(Hydroxymethyl)-3-(acetamidomethylene)succinate compound B, Tris-HCl, pH 8.0 Arthrobacter sp.
0.56
-
 2-(Hydroxymethyl)-3-(acetamidomethylene)succinate compound B, 2-(N-morpholino)ethane-sulfonic acid, pH 6.5 Arthrobacter sp.
0.68
-
 3-(acetamidomethylene)succinate compound A, 25°C, pH 7.0 Arthrobacter sp.

Metals/Ions

Metals/Ions Comment Organism Structure
additional information some inorganic cations activate Arthrobacter sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36600
-
 Cr-7, probably six identical subunits, 6 * 36600, SDS-PAGE Arthrobacter sp.
205000
-
 Cr-7, gel filtration Arthrobacter sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O Pseudomonas sp. compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O Arthrobacter sp. compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O Arthrobacter sp. involved in pyridoxine degradation ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O Pseudomonas sp. IA compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid ?
-
 ?
additional information Pseudomonas sp. enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen ?
-
 ?
additional information Arthrobacter sp. enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen ?
-
 ?
additional information Pseudomonas sp. IA enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen ?
-
 ?

Organism

Organism UniProt Comment Textmining
Arthrobacter sp.
-
 Cr-7
-
Pseudomonas sp.
-
 IA
-
Pseudomonas sp. IA
-
 IA
-

Purification (Commentary)

Purification (Comment) Organism
Cr-7 Arthrobacter sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Arthrobacter sp.
3.79
-
 25°C Arthrobacter sp.

Storage Stability

Storage Stability Organism
Cr-7, -20°C, 0.1 M Tris-HCl, pH 7.0, stable for several months Arthrobacter sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Pseudomonas sp. acetate + 2-(hydroxymethyl)-4-oxobutanoate + NH3 + CO2
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Arthrobacter sp. acetate + 2-(hydroxymethyl)-4-oxobutanoate + NH3 + CO2
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Pseudomonas sp. IA acetate + 2-(hydroxymethyl)-4-oxobutanoate + NH3 + CO2
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Pseudomonas sp. ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Arthrobacter sp. ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O involved in pyridoxine degradation Arthrobacter sp. ?
-
 ?
2-(hydroxymethyl)-3-(acetamidomethylene)succinate + H2O compound B, alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid Pseudomonas sp. IA ?
-
 ?
3-(acetamidomethylene)succinate + H2O compound A, alpha-(N-acetylaminomethylene)succinic acid: poor substrate Arthrobacter sp. acetate + 4-oxobutanoate + NH3 + CO2
-
 ?
additional information high substrate specificity Arthrobacter sp. ?
-
 ?
additional information enzyme highly specific, inducible amidohydrolase Arthrobacter sp. ?
-
 ?
additional information enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen Pseudomonas sp. ?
-
 ?
additional information enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen Arthrobacter sp. ?
-
 ?
additional information enzyme of vitamin B6 degradation, utilizing pyridoxine as sole source of carbon and nitrogen Pseudomonas sp. IA ?
-
 ?

Subunits

Subunits Comment Organism
multimer Cr-7, probably six identical subunits, 6 * 36600, SDS-PAGE Arthrobacter sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Arthrobacter sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
 pH 8.0, 1 min, stable at Arthrobacter sp.
100
-
 1 min, 65% activity retained Arthrobacter sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3
-
 2-(Hydroxymethyl)-3-(acetamidomethylene)succinate turnover no./subunit, 25°C Arthrobacter sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
 pI: 4.2, effect of buffer on pH optimum Arthrobacter sp.
6 7
-
 Arthrobacter sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 5 rapid loss of activity Arthrobacter sp.
6 10 stable at room temperature Arthrobacter sp.
8
-
 70°C, 1 min, stable at Arthrobacter sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.74
-
 Itaconic acid competitive, most effective inhibitor Arthrobacter sp.
20
-
 malonate competitive Arthrobacter sp.