Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.14.13.172 extracted from

  • Fang, T.; Zhou, N.Y.
    Purification and characterization of salicylate 5-hydroxylase, a three-component monooxygenase from Ralstonia sp. strain U2 (2014), Appl. Microbiol. Biotechnol., 98, 671-679.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Ralstonia sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05641
-
NADPH in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.
0.05976
-
NADH in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.
0.1071
-
salicylate in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.
0.3284
-
2-Nitrophenol in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.
0.9398
-
2,6-dihydroxybenzoate in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Iron the enzyme contains one Rieske-type [2Fe-2S] cluster and 7.5 g iron per mol enzyme Ralstonia sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
18800
-
3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence Ralstonia sp.
20000
-
3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE Ralstonia sp.
51000
-
3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence Ralstonia sp.
52000
-
3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE Ralstonia sp.
195200
-
gel filtration Ralstonia sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
salicylate + NADH + H+ + O2 Ralstonia sp.
-
2,5-dihydroxybenzoate + NAD+ + H2O
-
?
salicylate + NADH + H+ + O2 Ralstonia sp. U2
-
2,5-dihydroxybenzoate + NAD+ + H2O
-
?
salicylate + NADPH + H+ + O2 Ralstonia sp.
-
2,5-dihydroxybenzoate + NADP+ + H2O
-
?
salicylate + NADPH + H+ + O2 Ralstonia sp. U2
-
2,5-dihydroxybenzoate + NADP+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Ralstonia sp. O52379 and O52380 large oxygenase component and small oxagenase component
-
Ralstonia sp. U2 O52379 and O52380 large oxygenase component and small oxagenase component
-

Purification (Commentary)

Purification (Comment) Organism
the three-component Fe-S protein complex (NagAaGHAb) of the enzyme is purified by HisTrap column chromatography and Sephadex G-25 gel filtration Ralstonia sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
107.1
-
after 124fold purification, in 50 mM MES buffer (pH 6.4), at 22°C Ralstonia sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dihydroxybenzoate + NADH + H+ + O2
-
Ralstonia sp. ?
-
?
2,4-dihydroxybenzoate + NADH + H+ + O2
-
Ralstonia sp. U2 ?
-
?
2,6-dihydroxybenzoate + NADH + H+ + O2
-
Ralstonia sp. ?
-
?
2,6-dihydroxybenzoate + NADH + H+ + O2
-
Ralstonia sp. U2 ?
-
?
2-nitrophenol + NADH + H+ + O2
-
Ralstonia sp. ?
-
?
2-nitrophenol + NADH + H+ + O2
-
Ralstonia sp. U2 ?
-
?
salicylate + NADH + H+ + O2
-
Ralstonia sp. 2,5-dihydroxybenzoate + NAD+ + H2O
-
?
salicylate + NADH + H+ + O2
-
Ralstonia sp. U2 2,5-dihydroxybenzoate + NAD+ + H2O
-
?
salicylate + NADPH + H+ + O2
-
Ralstonia sp. 2,5-dihydroxybenzoate + NADP+ + H2O
-
?
salicylate + NADPH + H+ + O2
-
Ralstonia sp. U2 2,5-dihydroxybenzoate + NADP+ + H2O
-
?

Subunits

Subunits Comment Organism
heterohexamer 3 * 51000 + 3 * 18800, His6-tagged enzyme, calculated from amino acid sequence Ralstonia sp.
heterohexamer 3 * 52000 + 3 * 20000, His6-tagged enzyme, SDS-PAGE Ralstonia sp.

Synonyms

Synonyms Comment Organism
NagGH
-
Ralstonia sp.
S5H
-
Ralstonia sp.

Cofactor

Cofactor Comment Organism Structure
NADH
-
Ralstonia sp.
NADPH
-
Ralstonia sp.
[2Fe-2S]-center
-
Ralstonia sp.