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Literature summary for 1.14.13.6 extracted from

  • Ohta, Y.; Higgins, I.J.; Ribbons, D.W.
    Metabolism of resorcinylic compounds by bacteria. Purification and properties of orcinol hydroxylase from Pseudomonas putida 01 (1975), J. Biol. Chem., 250, 3814-3825.
    View publication on PubMed
Search for more literature for 1.14.13.6

Activating Compound

Activating Compound Comment Organism Structure
m-cresol increases activity Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
-
 Pseudomonas putida

General Stability

General Stability Organism
thiol reagents, e.g. 2-mercaptoethanol, stabilize Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
1,3-Dimethoxybenzene 45% inhibition at 1.7 mM Pseudomonas putida
2-Hydroxy-4-methoxybenzoate 68% inhibition at 1.7 mM Pseudomonas putida
2-Methylresorcinol 27% inhibition at 1.7 mM Pseudomonas putida
2-Nitroorcinol 64% inhibition at 1.7 mM Pseudomonas putida
3,4-dimethylphenol 54% inhibition at 1.7 mM Pseudomonas putida
3,5-Dihydroxybenzoate 59% inhibition at 1.7 mM Pseudomonas putida
3,5-Dimethylphenol 68% inhibition at 1.7 mM Pseudomonas putida
3-Hydroxybenzaldehyde 23% inhibition at 1.7 mM Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information
-
 Pseudomonas putida
0.03
-
 orcinol cofactor: NADH Pseudomonas putida
0.05
-
 orcinol cofactor: 3-acetylpyridine NADH Pseudomonas putida
0.07
-
 O2
-
 Pseudomonas putida
0.085
-
 orcinol cofactor: NADPH Pseudomonas putida
0.13
-
 NADH substrate: orcinol Pseudomonas putida
0.19
-
 resorcinol
-
 Pseudomonas putida
0.65
-
 3-acetylpyridine-NADH substrate: orcinol Pseudomonas putida
2.5
-
 NADH substrate: resorcinol Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
63000
-
 gel filtration Pseudomonas putida
65000
-
 sedimentation equilibrium Pseudomonas putida
68000
-
 1 * 68000, SDS-PAGE Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
 01
-
Pseudomonas putida 1
-
 01
-

Purification (Commentary)

Purification (Comment) Organism
using protamine sulfate treatment, DEAE-cellulose column chromatography, ammonium sulfate treatment and column chromatography on Sephadex G-100 followed by ammonium sulfate treatment and column chromatography on hydroxylapatite Pseudomonas putida

Renatured (Commentary)

Renatured (Comment) Organism
most of the activity of the apoenzyme is reconstituted by the addition of FAD, FMN is a poor substitute for FAD Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.8
-
 apoenzyme without flavin addition Pseudomonas putida
3.5
-
 apoenzyme with addition of FMN Pseudomonas putida
14.7
-
 apoenzyme with addition of FAD Pseudomonas putida
18
-
 holoenzyme Pseudomonas putida
24
-
-
 Pseudomonas putida

Storage Stability

Storage Stability Organism
4°C, stabilized by thiol reagents, e.g. 2-mercaptoethanol, some days Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-cresol + NADH + O2 nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated Pseudomonas putida 3-methylcatechol + NAD+ + H2O
-
 ?
3-cresol + NADH + O2 nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated Pseudomonas putida 1 3-methylcatechol + NAD+ + H2O
-
 ?
3-ethylphenol + NADH + O2 no substrate, mimic orcinol as effector Pseudomonas putida ?
-
 ?
3-ethylphenol + NADH + O2 no substrate, mimic orcinol as effector Pseudomonas putida 1 ?
-
 ?
4-bromoresorcinol + NADH + O2
-
 Pseudomonas putida ?
-
 ?
4-methylresorcinol + NADH + O2
-
 Pseudomonas putida ?
-
 ?
additional information reduced orcinol hydroxylase is able to transfer reducing equivalents to a variety of electron acceptors, other than oxygen. These include free FAD, ferricyanide, cytochrome c, acetylpyridine-NAD and tetrazolium salts Pseudomonas putida ?
-
 ?
additional information reduced orcinol hydroxylase is able to transfer reducing equivalents to a variety of electron acceptors, other than oxygen. These include free FAD, ferricyanide, cytochrome c, acetylpyridine-NAD and tetrazolium salts Pseudomonas putida 1 ?
-
 ?
orcinol + 3-acetylpyridine-NADH + H+ + O2
-
 Pseudomonas putida 2,3,5-trihydroxytoluene + 3-acetylpyridine-NAD+ + H2O
-
 r
orcinol + NADH + O2
-
 Pseudomonas putida 2,3,5-trihydroxytoluene + NAD+ + H2O
-
 ?
orcinol + NADH + O2
-
 Pseudomonas putida 1 2,3,5-trihydroxytoluene + NAD+ + H2O
-
 ?
resorcinol + NADH + O2
-
 Pseudomonas putida hydroxyquinol + NAD+ + H2O
-
 ?
resorcinol + NADH + O2
-
 Pseudomonas putida 1 hydroxyquinol + NAD+ + H2O
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 68000, SDS-PAGE Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
26
-
 orcinol crystalline enzyme Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
 difficult to evaluate, because the values do not take into account the nonenzymic oxidation rate of the product of the reaction Pseudomonas putida
6.8
-
 assay at Pseudomonas putida

pH Stability

pH Stability pH Stability Maximum Comment Organism
7
-
 highest stability Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
3-Acetyl pyridine nucleotide reduced Pseudomonas putida
FAD flavoprotein Pseudomonas putida
FAD 1 mol of FAD per mol of protein Pseudomonas putida
NADH
-
 Pseudomonas putida
NADPH
-
 Pseudomonas putida