BRENDA - Enzyme Database show
show all sequences of 1.14.13.135

Biodegradation of phenanthrene by Alcaligenes sp. strain PPH: partial purification and characterization of 1-hydroxy-2-naphthoic acid hydroxylase

Deveryshetty, J.; Phale, P.S.; FEMS Microbiol. Lett. 311, 93-101 (2010)

Data extracted from this reference:

General Stability
General Stability
Organism
1-hydroxy-2-naphthoic acid hydroxylase in the cell-free extract is stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol
Alcaligenes sp.
repeated freezing and thawing led to inactivation of the enzyme
Alcaligenes sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0716
-
1-hydroxy-2-naphthoate
pH 7.5, 30C, with NADH
Alcaligenes sp.
0.0755
-
1-hydroxy-2-naphthoate
pH 7.5, 30C, with NADPH
Alcaligenes sp.
0.0846
-
NADPH
pH 7.5, 30C
Alcaligenes sp.
0.087
-
NADH
pH 7.5, 30C
Alcaligenes sp.
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
no effect by 1 mM of Fe2+, Fe3+, Mg2+, Mn2+, Ca2+, Zn2+, and Cu2+, and by metal chelators, such as EDTA, 2,2-dipyridyl and 1',10'-phenanthroline, at 1 mM
Alcaligenes sp.
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
2 * 34000, SDS-PAGE
Alcaligenes sp.
66000
-
gel filtration
Alcaligenes sp.
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1-hydroxy-2-naphthoate + NADH + O2
Alcaligenes sp.
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
?
1-hydroxy-2-naphthoate + NADPH + O2
Alcaligenes sp.
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
1,2-dihydroxynaphthalene + NADP+ + H2O + CO2
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Alcaligenes sp.
-
phenanthrene-degrading
-
no activity in Pseudomonas putida
-
-
-
no activity in Pseudomonas putida CSV86
-
-
-
Purification (Commentary)
Commentary
Organism
native enzyme 81fold by heat treatment at 60C, ammonium sulfate fractionation and anion exchange chromatography. Additional purification steps, such as hydrophobic interaction chromatography or gel filtration, lead to the total or a significant, 70%, loss of activity, respectively, without achieving any further purification
Alcaligenes sp.
Source Tissue
Source Tissue
Commentary
Organism
Textmining
culture condition:phenanthrene-grown cell
-
Alcaligenes sp.
-
culture condition:salicylate-grown cell
significantly lower activity compared to phenanthrene-grown cells
Alcaligenes sp.
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.0031
-
salicylate-grown cells, pH 7.5, 30C
Alcaligenes sp.
0.0877
-
phenanthrene-grown cells, pH 7.5, 30C
Alcaligenes sp.
25.3
-
purified enzyme, cofactor NADPH, pH 7.5, 30C
Alcaligenes sp.
Storage Stability
Storage Stability
Organism
-20C, purified enzyme, stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol, retains 100% activity, after 60 days
Alcaligenes sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-hydroxy-2-naphthoate + NADH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
-
?
1-hydroxy-2-naphthoate + NADH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
-
?
1-hydroxy-2-naphthoate + NADPH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NADP+ + H2O + CO2
-
-
-
?
additional information
substrate specificity, overview. No activity on salicylic acid, 3-hydroxy-2-naphthoic acid, 2-hydroxy-1-naphthoic acid, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 1-naphthol, 2-naphthol, 1-naphthoic acid, 2-naphthoic acid, gentisic acid or catechol
712039
Alcaligenes sp.
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
homodimer
2 * 34000, SDS-PAGE
Alcaligenes sp.
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
assay at
Alcaligenes sp.
Temperature Stability [C]
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
60
-
5 min, cell-free extract, the enzyme is stable in presence of 1-hydroxy-2-naphthoic acid
Alcaligenes sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
-
Alcaligenes sp.
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
holoenzyme contains FAD. Km is 0.0047 mM. Preparation of the apoenzyme by the acid-ammonium sulfate, at 2 M, dialysis method, the apoenzyme is colorless, inactive and loses the characteristic flavin absorption spectra but regains about 90% of maximal activity when reconstituted with FAD
Alcaligenes sp.
additional information
FMN is a poor substitute for FAD
Alcaligenes sp.
NADH
Vmax/Km is similar for NADPH and NADH
Alcaligenes sp.
NADPH
Vmax/Km is similar for NADPH and NADH
Alcaligenes sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
holoenzyme contains FAD. Km is 0.0047 mM. Preparation of the apoenzyme by the acid-ammonium sulfate, at 2 M, dialysis method, the apoenzyme is colorless, inactive and loses the characteristic flavin absorption spectra but regains about 90% of maximal activity when reconstituted with FAD
Alcaligenes sp.
additional information
FMN is a poor substitute for FAD
Alcaligenes sp.
NADH
Vmax/Km is similar for NADPH and NADH
Alcaligenes sp.
NADPH
Vmax/Km is similar for NADPH and NADH
Alcaligenes sp.
General Stability (protein specific)
General Stability
Organism
1-hydroxy-2-naphthoic acid hydroxylase in the cell-free extract is stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol
Alcaligenes sp.
repeated freezing and thawing led to inactivation of the enzyme
Alcaligenes sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0716
-
1-hydroxy-2-naphthoate
pH 7.5, 30C, with NADH
Alcaligenes sp.
0.0755
-
1-hydroxy-2-naphthoate
pH 7.5, 30C, with NADPH
Alcaligenes sp.
0.0846
-
NADPH
pH 7.5, 30C
Alcaligenes sp.
0.087
-
NADH
pH 7.5, 30C
Alcaligenes sp.
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
no effect by 1 mM of Fe2+, Fe3+, Mg2+, Mn2+, Ca2+, Zn2+, and Cu2+, and by metal chelators, such as EDTA, 2,2-dipyridyl and 1',10'-phenanthroline, at 1 mM
Alcaligenes sp.
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
2 * 34000, SDS-PAGE
Alcaligenes sp.
66000
-
gel filtration
Alcaligenes sp.
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1-hydroxy-2-naphthoate + NADH + O2
Alcaligenes sp.
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
?
1-hydroxy-2-naphthoate + NADPH + O2
Alcaligenes sp.
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
1,2-dihydroxynaphthalene + NADP+ + H2O + CO2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
native enzyme 81fold by heat treatment at 60C, ammonium sulfate fractionation and anion exchange chromatography. Additional purification steps, such as hydrophobic interaction chromatography or gel filtration, lead to the total or a significant, 70%, loss of activity, respectively, without achieving any further purification
Alcaligenes sp.
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
culture condition:phenanthrene-grown cell
-
Alcaligenes sp.
-
culture condition:salicylate-grown cell
significantly lower activity compared to phenanthrene-grown cells
Alcaligenes sp.
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.0031
-
salicylate-grown cells, pH 7.5, 30C
Alcaligenes sp.
0.0877
-
phenanthrene-grown cells, pH 7.5, 30C
Alcaligenes sp.
25.3
-
purified enzyme, cofactor NADPH, pH 7.5, 30C
Alcaligenes sp.
Storage Stability (protein specific)
Storage Stability
Organism
-20C, purified enzyme, stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol, retains 100% activity, after 60 days
Alcaligenes sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-hydroxy-2-naphthoate + NADH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
-
?
1-hydroxy-2-naphthoate + NADH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NAD+ + H2O + CO2
-
-
-
?
1-hydroxy-2-naphthoate + NADPH + O2
a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid
712039
Alcaligenes sp.
1,2-dihydroxynaphthalene + NADP+ + H2O + CO2
-
-
-
?
additional information
substrate specificity, overview. No activity on salicylic acid, 3-hydroxy-2-naphthoic acid, 2-hydroxy-1-naphthoic acid, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 1-naphthol, 2-naphthol, 1-naphthoic acid, 2-naphthoic acid, gentisic acid or catechol
712039
Alcaligenes sp.
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 34000, SDS-PAGE
Alcaligenes sp.
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
assay at
Alcaligenes sp.
Temperature Stability [C] (protein specific)
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
60
-
5 min, cell-free extract, the enzyme is stable in presence of 1-hydroxy-2-naphthoic acid
Alcaligenes sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
-
Alcaligenes sp.
Expression
Organism
Commentary
Expression
Alcaligenes sp.
the enzyme is induced by growth on phenanthrene
up
General Information
General Information
Commentary
Organism
metabolism
in the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
Alcaligenes sp.
General Information (protein specific)
General Information
Commentary
Organism
metabolism
in the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway
Alcaligenes sp.
Expression (protein specific)
Organism
Commentary
Expression
Alcaligenes sp.
the enzyme is induced by growth on phenanthrene
up
Other publictions for EC 1.14.13.135
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
715057
Nayak
A catabolic pathway for the de ...
Pseudoxanthomonas sp., Pseudoxanthomonas sp. PNK-04
FEMS Microbiol. Lett.
320
128-134
2011
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-
-
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-
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2
-
3
-
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3
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2
-
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-
-
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-
2
-
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3
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2
-
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-
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-
-
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712039
Deveryshetty
Biodegradation of phenanthrene ...
Alcaligenes sp., no activity in Pseudomonas putida, no activity in Pseudomonas putida CSV86
FEMS Microbiol. Lett.
311
93-101
2010
-
-
-
-
-
2
-
4
-
1
2
2
-
4
-
-
1
-
-
2
3
1
4
1
1
-
1
-
1
-
-
4
-
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-
4
-
-
2
-
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-
4
-
1
2
2
-
-
-
1
-
2
3
1
4
1
1
-
1
-
1
-
-
-
1
1
1
1
-
-