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Literature summary extracted from

  • Jouanneau, Y.; Meyer, C.; Jakoncic, J.; Stojanoff, V.; Gaillard, J.
    Characterization of a naphthalene dioxygenase endowed with an exceptionally broad substrate specificity toward polycyclic aromatic hydrocarbons (2006), Biochemistry, 45, 12380-12391.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.12.12 expressed in Pseudomonas putida Sphingomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.12.12 0.00042
-
 biphenyl at 30°C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0 Sphingomonas sp.
1.14.12.12 0.00092
-
 Naphthalene at 30°C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0 Sphingomonas sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.12.12 Fe2+ the iron content of the oxygenase component ht-PhnI varies between 1.73 and 2.55 Fe atoms per pair of subunits depending on preparation, preincubation of ht-PhnI with ferrous ions under reducing conditions prior to enzyme assay results in a marginal increase of activity, the iron content of the ferredoxin component ht-PhnA3 is estimated to be 1.5 mol/mol of ferredoxin Sphingomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.12.12 13500
-
 ferredoxin component, gel filtration Sphingomonas sp.
1.14.12.12 20000
-
 3 * 52000, 3 * 20000, alpha3beta3 hexamer, oxygenase component, SDS-PAGE Sphingomonas sp.
1.14.12.12 45000
-
 reductase component, gel filtration Sphingomonas sp.
1.14.12.12 52000
-
 3 * 52000, 3 * 20000, alpha3beta3 hexamer, oxygenase component, SDS-PAGE Sphingomonas sp.
1.14.12.12 200000
-
 oxydase component, gel filtration Sphingomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
1.14.12.12 Sphingomonas sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.12.12 DEAE-cellulose DE52 column chromatography, immobilized metal affinity chromatography and DEAE cellulose column chromatography Sphingomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.12.12 3 benz[a]anthracene + 3 NADH + 3 O2 + 3 H+
-
 Sphingomonas sp. benz[a]anthracene-10,11-cis-dihydrodiol + benz[a]anthracene-8,9-cis-dihydrodiol + benz[a]anthracene-1,2-cis-dihydrodiol + 3 NAD+
-
 ?
1.14.12.12 benz[a]anthracene-1,2-cis-dihydrodiol + NADH + O2
-
 Sphingomonas sp. benz[a]anthracene-1,2,10,11-bis-cis-dihydrodiol + NAD+
-
 ?
1.14.12.12 benz[a]anthracene-10,11-cis-dihydrodiol + NADH + O2
-
 Sphingomonas sp. benz[a]anthracene-1,2,10,11-bis-cis-dihydrodiol + NAD+
-
 ?
1.14.12.12 biphenyl + NADH + O2
-
 Sphingomonas sp. ? + NAD+
-
 ?
1.14.12.12 chrysene + NADH + O2
-
 Sphingomonas sp. chrysene-3,4-cis-dihydrodiol + chrysene-3,4,9,10-bis-cis-dihydrodiol + NAD+
-
 ?
1.14.12.12 naphthalene + NADH + O2
-
 Sphingomonas sp. (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
-
 ?
1.14.12.12 phenanthrene + NADH + O2
-
 Sphingomonas sp. ? + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.12.12 hexamer 3 * 52000, 3 * 20000, alpha3beta3 hexamer, oxygenase component, SDS-PAGE Sphingomonas sp.
1.14.12.12 monomer 1 * 13500, ferredoxin component, gel filtration Sphingomonas sp.

Synonyms

EC Number Synonyms Comment Organism
1.14.12.12 naphthalene dioxygenase the enzyme is a three-component dioxygenase complex consisting of ht-PhnI (oxygenase component), ht-PhnA3 (ferredoxin component), and ht-PhnA4 (reductase component) Sphingomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.12.12 1.01
-
 biphenyl at 30°C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0 Sphingomonas sp.
1.14.12.12 1.82
-
 Naphthalene at 30°C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0 Sphingomonas sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.12.12 NADH
-
 Sphingomonas sp.