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Literature summary extracted from

  • Suske, W.A.; Held, M.; Schmid, A.; Fleischmann, T.; Wubbolts, M.G.; Kohler, H.P.E.
    Purification and characterization of 2-hydroxybiphenyl 3-monooxygenase, a novel NADH-dependent, FAD-containing aromatic hydroxylase from Pseudomonas azelaica HBP1 (1997), J. Biol. Chem., 272, 24257-24265.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.44 2,3-Dihydroxybiphenyl inhibits reaction with 2-hydroxybiphenyl Pseudomonas nitroreducens
1.14.13.44 AgNO3 0.01 mM, complete inhibition Pseudomonas nitroreducens
1.14.13.44 CuSO4 0.01 mM, complete inhibition Pseudomonas nitroreducens
1.14.13.44 FeSO4 0.08 mM, 30% inhibition Pseudomonas nitroreducens
1.14.13.44 HgCl2 0.01 mM, complete inhibition Pseudomonas nitroreducens
1.14.13.44 NaCl 10 mM, 36% inhibition. 100 mM, 89% inhibition Pseudomonas nitroreducens
1.14.13.44 p-hydroxymercuribenzoate partial protection in presence of 2-hydroxybiphenyl, reversed by excess of dithiothreitol Pseudomonas nitroreducens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.44 0.0028
-
2-Hydroxybiphenyl reaction with NADH and O2 Pseudomonas nitroreducens
1.14.13.44 0.0031
-
2-Hydroxybiphenyl reaction with NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 0.0034
-
2,2'-Dihydroxybiphenyl reaction with NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 0.004
-
2,2'-Dihydroxybiphenyl reaction with NADH and O2 Pseudomonas nitroreducens
1.14.13.44 0.0057
-
2-sec-Butylphenol reaction with NADH or NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 0.0216
-
NADH reaction with 2,2'-dihydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 0.0268
-
NADH reaction with 2-hydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 0.0292
-
O2 reaction with 2-hydroxybiphenyl and NADH Pseudomonas nitroreducens
1.14.13.44 0.0943
-
NADPH reaction with 2,2'-dihydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 0.137
-
NADPH reaction with 2-hydroxybiphenyl and O2 Pseudomonas nitroreducens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.44 60000
-
4 * 60000, SDS-PAGE Pseudomonas nitroreducens
1.14.13.44 256000
-
gel filtration Pseudomonas nitroreducens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.44 2-hydroxybiphenyl + NADH + O2 Pseudomonas nitroreducens first enzyme of 2-hydroxybiphenyl degradation 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADH + O2 Pseudomonas nitroreducens HBP1 first enzyme of 2-hydroxybiphenyl degradation 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.44 Pseudomonas nitroreducens
-
HBP1
-
1.14.13.44 Pseudomonas nitroreducens HBP1
-
HBP1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.44 native and recombinant enzyme Pseudomonas nitroreducens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.44 additional information
-
-
Pseudomonas nitroreducens

Storage Stability

EC Number Storage Stability Organism
1.14.13.44 -20°C, pure enzyme at concentration of 3.8 mg/ml in 50 mM phosphate buffer, pH 7.5, stable for at least 6 months Pseudomonas nitroreducens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.44 2,2'-dihydroxybiphenyl + NADH + O2
-
Pseudomonas nitroreducens 2,2',3-trihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2,2'-dihydroxybiphenyl + NADPH + O2
-
Pseudomonas nitroreducens 2,2',3-trihydroxybiphenyl + NADP+ + H2O
-
?
1.14.13.44 2,5-dihydroxybiphenyl + NADH + O2
-
Pseudomonas nitroreducens 2,3,5-trihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-ethylphenol + NADH + O2
-
Pseudomonas nitroreducens 1,2-dihydroxy-3-ethylbenzene + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADH + O2
-
Pseudomonas nitroreducens 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADH + O2 first enzyme of 2-hydroxybiphenyl degradation Pseudomonas nitroreducens 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADH + O2
-
Pseudomonas nitroreducens HBP1 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADH + O2 first enzyme of 2-hydroxybiphenyl degradation Pseudomonas nitroreducens HBP1 2,3-dihydroxybiphenyl + NAD+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADPH + O2
-
Pseudomonas nitroreducens 2,3-dihydroxybiphenyl + NADP+ + H2O
-
?
1.14.13.44 2-hydroxybiphenyl + NADPH + O2
-
Pseudomonas nitroreducens HBP1 2,3-dihydroxybiphenyl + NADP+ + H2O
-
?
1.14.13.44 2-methylphenol + NADH + O2
-
Pseudomonas nitroreducens 1,2-dihydroxy-3-methylbenzene + NAD+ + H2O
-
?
1.14.13.44 2-propylphenol + NADH + O2
-
Pseudomonas nitroreducens 1,2-dihydroxy-3-propylbenzene + NAD+ + H2O
-
?
1.14.13.44 2-propylphenol + NADH + O2
-
Pseudomonas nitroreducens HBP1 1,2-dihydroxy-3-propylbenzene + NAD+ + H2O
-
?
1.14.13.44 2-sec-butylphenol + NADH + O2
-
Pseudomonas nitroreducens 2-sec-butylcatechol + NAD+ + H2O
-
?
1.14.13.44 2-sec-butylphenol + NADPH + O2
-
Pseudomonas nitroreducens 2-sec-butylcatechol + NADP+ + H2O
-
?
1.14.13.44 2-tert-butylphenol + NADH + O2
-
Pseudomonas nitroreducens 1,2-dihydroxy-3-tert-butylbenzene + NAD+ + H2O
-
?
1.14.13.44 additional information the substrates partially uncouple oxygen activation from hydroxylation with resultant reduction of both atoms of oxygen to form hydrogen peroxide Pseudomonas nitroreducens ?
-
?
1.14.13.44 additional information the substrates partially uncouple oxygen activation from hydroxylation with resultant reduction of both atoms of oxygen to form hydrogen peroxide Pseudomonas nitroreducens HBP1 ?
-
?

Subunits

EC Number Subunits Comment Organism
1.14.13.44 tetramer 4 * 60000, SDS-PAGE Pseudomonas nitroreducens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.44 9
-
2,2'-Dihydroxybiphenyl reaction with NADH and O2 Pseudomonas nitroreducens
1.14.13.44 9.4
-
2,2'-Dihydroxybiphenyl reaction with NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 9.8
-
NADH reaction with 2,2'-dihydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 10.2
-
2-sec-Butylphenol reaction with NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 11.2
-
NADPH reaction with 2,2'-dihydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 12.8
-
2-Hydroxybiphenyl reaction with NADPH and O2 Pseudomonas nitroreducens
1.14.13.44 14.9
-
2-Hydroxybiphenyl reaction with NADH and O2 Pseudomonas nitroreducens
1.14.13.44 15.8
-
2-sec-Butylphenol reaction with NADH and O2 Pseudomonas nitroreducens
1.14.13.44 16.2
-
NADH reaction with 2-hydroxybiphenyl and O2 Pseudomonas nitroreducens
1.14.13.44 16.2
-
O2 reaction with 2-hydroxybiphenyl and NADH Pseudomonas nitroreducens
1.14.13.44 18.8
-
NADPH reaction with 2-hydroxybiphenyl and O2 Pseudomonas nitroreducens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.44 7.5
-
-
Pseudomonas nitroreducens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.13.44 7.2 7.8 more than 80% of maximal activity at pH 7.2 and pH 7.8, beyond pH 7.8 activity declines abruptly with increasing pH Pseudomonas nitroreducens

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.44 FAD one molecule of noncovalently bound FAD per subunit Pseudomonas nitroreducens
1.14.13.44 NADH
-
Pseudomonas nitroreducens
1.14.13.44 NADPH can replace NADH as electron donor, Km-value for NADPH is much higher than for NADH Pseudomonas nitroreducens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.13.44 0.9
-
2,3-Dihydroxybiphenyl
-
Pseudomonas nitroreducens