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EC Tree
IUBMB Comments The enzyme from Pseudomonas diminuta is specific towards N-containing N-heterocyclic substrates, including isoquinoline, isoquinolin-5-ol, phthalazine and quinazoline. Electron acceptors include 1,2-benzoquinone, cytochrome c, ferricyanide, iodonitrotetrazolium chloride, nitroblue tetrazolium, Meldola blue and phenazine methosulfate.
The enzyme appears in viruses and cellular organisms
Synonyms
isoquinoline 1-oxidoreductase,
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Isoquinoline hydroxylase
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Isoquinoline oxidase
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Reductase, isoquinoline
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IOR
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Isoquinoline + acceptor + H2O = isoquinolin-1(2H)-one + reduced acceptor
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Isoquinoline:acceptor 1-oxidoreductase (hydroxylating)
The enzyme from Pseudomonas diminuta is specific towards N-containing N-heterocyclic substrates, including isoquinoline, isoquinolin-5-ol, phthalazine and quinazoline. Electron acceptors include 1,2-benzoquinone, cytochrome c, ferricyanide, iodonitrotetrazolium chloride, nitroblue tetrazolium, Meldola blue and phenazine methosulfate.
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5-hydroxyisoquinoline + acceptor + H2O
1,5-dihydroisoquinoline + reduced acceptor
isoquinoline + 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride + H2O
isoquinolin-1(2H)-one + reduced 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride
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r
isoquinoline + acceptor + H2O
1-oxo-1,2-dihydroisoquinoline + reduced acceptor
isoquinoline + acceptor + H2O
1-oxoisoquinoline + reduced acceptor
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acceptor: ferricyanide, 1,4-benzoquinone, cytochrome c, 2,6-dichloroindophenol,2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride, phenazine methosulfate, thionine, nitroblue tetrazolium, Meldola's blue
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Isoquinoline + acceptor + H2O
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phthalazine + acceptor + H2O
1-oxophthalazine + reduced acceptor
quinazoline + acceptor + H2O
4-hydroxyquinazoline + reduced acceptor
quinazoline + acceptor + H2O
4-oxoquinazoline + reduced acceptor
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5-hydroxyisoquinoline + acceptor + H2O
1,5-dihydroisoquinoline + reduced acceptor
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5-hydroxyisoquinoline + acceptor + H2O
1,5-dihydroisoquinoline + reduced acceptor
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isoquinoline + acceptor + H2O
1-oxo-1,2-dihydroisoquinoline + reduced acceptor
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acceptors: 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride, formazan, medola's blue, N-methyldibenzylpyrazine methylsulfate, nitroblue tetrazolium chloride, cytochrome c, ferricyanide, 1,2-benzoquinone
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isoquinoline + acceptor + H2O
1-oxo-1,2-dihydroisoquinoline + reduced acceptor
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acceptors: 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride, formazan, medola's blue, N-methyldibenzylpyrazine methylsulfate, nitroblue tetrazolium chloride, cytochrome c, ferricyanide, 1,2-benzoquinone
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Isoquinoline + acceptor + H2O
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first enzyme of the isoquinoline catabolism
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Isoquinoline + acceptor + H2O
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first enzyme of the isoquinoline catabolism
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phthalazine + acceptor + H2O
1-oxophthalazine + reduced acceptor
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phthalazine + acceptor + H2O
1-oxophthalazine + reduced acceptor
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at 89% of the activity compared to isoquinoline
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phthalazine + acceptor + H2O
1-oxophthalazine + reduced acceptor
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at 89% of the activity compared to isoquinoline
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quinazoline + acceptor + H2O
4-hydroxyquinazoline + reduced acceptor
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at 45% of the activity compared to isoquinoline
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quinazoline + acceptor + H2O
4-hydroxyquinazoline + reduced acceptor
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at 45% of the activity compared to isoquinoline
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Isoquinoline + acceptor + H2O
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Isoquinoline + acceptor + H2O
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first enzyme of the isoquinoline catabolism
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Isoquinoline + acceptor + H2O
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first enzyme of the isoquinoline catabolism
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iron-sulfur centre
two distinct 2Fe-2S clusters
molybdopterin cytosine dinucleotide
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Pterin molybdenum cofactor
additional information
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enzyme contains no FAD
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Pterin molybdenum cofactor
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Pterin molybdenum cofactor
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the enzyme contains a pterin molybdenum cofactor with a monooxo-monosulfido-type molybdenum center
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Fe2+
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two (2Fe+2S) clusters
Iron
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enzyme contains 3.95 g-atom of iron per mol of enzyme. The iron and the acid-labile sulfur are arranged in two (2Fe-2S) clusters
Mo
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enzyme contains 0.85 g-atom of molybdenum per mol of enzyme
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5-Hydroxyisoquinoline
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rapid and very effective inactivation in presence as well as in absence of the electron acceptors iodonitrotetrazolium chloride, phenazine methosulfate or ferricyanide
Isoquinoline
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slow inhibition in absence of an electron acceptor
p-hydroxymercuribenzoate
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1.34
2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride
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with isoquinoline as cosubstrate
0.016
Isoquinoline
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with 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-tetrazolium chloride as cosubstrate
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brenda
alpha-subunit
Swissprot
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beta-subunit
Swissprot
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alpha-subunit
Swissprot
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beta-subunit
Swissprot
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possibly associated with
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possibly associated with
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16000
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1 * 80000 + 1 * 16000, SDS-PAGE
16399
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1 * 16399 + 1 * 84429, calculation from nucleotide sequence
80000
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1 * 80000 + 1 * 16000, SDS-PAGE
84429
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1 * 16399 + 1 * 84429, calculation from nucleotide sequence
93000 - 95000
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gel filtration, analytical ultracentrifugation
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dimer
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1 * 80000 + 1 * 16000, SDS-PAGE
dimer
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1 * 16399 + 1 * 84429, calculation from nucleotide sequence
dimer
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1 * 80000 + 1 * 16000, SDS-PAGE
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dimer
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1 * 16399 + 1 * 84429, calculation from nucleotide sequence
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by the hanging-drop vapour-diffusion method, using two different crystallization buffers, streak-seeding and cross-linking are essential to obtain well diffracting crystals
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additional information
expression of recombinant enzyme in Pseudomonas putida as a catalytically active enzyme requires the presence of quinoline and accessory gene(s) encoding product(s) essential for the assembly of enzyme or which are part of a quinoline regulon
additional information
expression of recombinant enzyme in Pseudomonas putida as a catalytically active enzyme requires the presence of quinoline and accessory gene(s) encoding product(s) essential for the assembly of enzyme or which are part of a quinoline regulon
additional information
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expression of recombinant enzyme in Pseudomonas putida as a catalytically active enzyme requires the presence of quinoline and accessory gene(s) encoding product(s) essential for the assembly of enzyme or which are part of a quinoline regulon
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60
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5 min, rapid decrease of activity above
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by two cation-exchange chromatography steps, applying Triton X-100 overcomes retention problems of IOR in the first cation-exchange chromatography step
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including Triton X-100 in a precipitation step
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comparative sequence analysis of the iorA and iorB genes
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Stephan, I.; Tshisuaka, B.; Fetzner, S.; Lingens, F.
Quinaldine 4-oxidase from Arthrobacter sp. Ru61a, a versatile procaryotic molybdenum-containing hydroxylase active towards N-containing heterocyclic compounds and aromatic aldehydes
Eur. J. Biochem.
236
155-162
1996
Brevundimonas diminuta
brenda
Lehmann, M.; Tshisuaka, B.; Fetzner, S.; Roger, P.; Lingens, F.
Purification and characterization of isoquinoline 1-oxidoreductase from Pseudomonas diminuata 7, a novel molybdenum-containing hydroxylase
J. Biol. Chem.
269
11254-11260
1994
Brevundimonas diminuta, Brevundimonas diminuta 7
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Lehmann, M.; Tshisuaka, B.; Fetzner, S.; Lingens, F.
Molecular cloning of the isoquinoline 1-oxidoreductase genes from Pseudomonas diminuata 7, structural analysis of IorA and IorB, and sequence comparisons with other molybdenum-containing hydroxylases
J. Biol. Chem.
270
14420-14429
1995
Brevundimonas diminuta, Brevundimonas diminuta 7
brenda
Boer, D.R.; Mueller, A.; Fetzner, S.; Lowe, D.J.; Romao, M.J.
On the purification and preliminary crystallographic analysis of isoquinoline 1-oxidoreductase from Brevundimonas diminuta 7
Acta Crystallogr. Sect. F
F61
137-140
2005
Brevundimonas diminuta, Brevundimonas diminuta 7
brenda
Israel, I.; Sohni, M.; Fetzner, S.
Expression of the iorAB genes from Brevundimonas diminuta 7 encoding the molybdenum hydroxylase isoquinoline 1-oxidoreductase in Pseudomonas putida
FEMS Microbiol. Lett.
210
123-127
2002
Brevundimonas diminuta (Q51697), Brevundimonas diminuta (Q51698), Brevundimonas diminuta 7 (Q51697), Brevundimonas diminuta 7 (Q51698), Brevundimonas diminuta 7
brenda
Boer, D.R.; Mueller, A.; Fetzner, S.; Lowe, D.J.; Romao, M.J.
On the purification and preliminary crystallographic analysis of isoquinoline 1-oxidoreductase from Brevundimonas diminuta 7
Acta Crystallogr. Sect. F
61
137-140
2005
Brevundimonas diminuta, Brevundimonas diminuta 7
brenda
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