Information on EC 1.14.12.12 - naphthalene 1,2-dioxygenase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY hide
1.14.12.12
-
RECOMMENDED NAME
GeneOntology No.
naphthalene 1,2-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Ethylbenzene degradation
-
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Metabolic pathways
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Microbial metabolism in diverse environments
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Naphthalene degradation
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naphthalene degradation (aerobic)
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Polycyclic aromatic hydrocarbon degradation
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SYSTEMATIC NAME
IUBMB Comments
naphthalene,NADH:oxygen oxidoreductase (1,2-hydroxylating)
This enzyme is a member of the ring-hydroxylating dioxygenase (RHD) family of bacterial enzymes that play a critical role in the degradation of aromatic compounds, such as polycyclic aromatic hydrocarbons [5]. This enzyme comprises a multicomponent system, containing a reductase that is an iron-sulfur flavoprotein (FAD; EC 1.18.1.3, ferredoxin---NAD+ reductase), an iron-sulfur oxygenase, and ferredoxin. Requires Fe2+.
CAS REGISTRY NUMBER
COMMENTARY hide
9074-04-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
grown on naphthalene as sole carbon source
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Manually annotated by BRENDA team
Escherichia coli JM109(DE3)
strain JM109(DE3)
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-
Manually annotated by BRENDA team
strain NCIMB 40531
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Manually annotated by BRENDA team
strain 119, unable to grow on naphthalene
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Manually annotated by BRENDA team
8859
-
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Manually annotated by BRENDA team
strain HKT554
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Manually annotated by BRENDA team
strain HOB1
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Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9616-4
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-
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Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9816
strain NCIB 9816
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Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
strain NCIMB12038
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Manually annotated by BRENDA team
strain NCIMB12038
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Manually annotated by BRENDA team
strain CHY-1
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Manually annotated by BRENDA team
strain CHY-1
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(1S)-indanol + NADH + O2
(1S)-indanol + trans-1,3-indandiol
show the reaction diagram
-
recombinant enzyme
85.5% indanol, 11.5% 1,3-indandiol, minor products: (1S)-indenol and 1-indanone
?
(2S)-flavanone + NADH + H+ + O2
8-hydroxy-(2S)-flavanone + NAD+ + ?
show the reaction diagram
(3S)-isoflavanone + NADH + H+ + O2
8-hydroxy-(3S)-isoflavanone + NAD+ + ?
show the reaction diagram
(R)-1-indanol + NAD+ + O2
cis-1,3-indandiol + (R)-3-hydroxy-1-indanone + cis-1,2,3-indantriol + NAD+
show the reaction diagram
-
activity in E. coli cells expressing recombinant naphthalene dioxygenase
71% cis-1,3-indandiol, 18.2% (R)-3-hydroxy-1-indanone and 10.8% cis-1,2,3-indantriol
?
(S)-1-indanol + NAD+ + O2
trans-(1S,3S)-indan-1,3-diol + (R)-3-hydroxy-1-indanone + NAD+
show the reaction diagram
-
activity in E. coli cells expressing recombinant naphthalene dioxygenase
95.5% trans-(1S,3S)-indan-1,3-diol
?
(S)-1-indenol + NADH + O2
syn-2,3-dihydroxy-1-indanol + anti-2,3-dihydroxy-1-indanol
show the reaction diagram
-
recombinant enzyme system
-
?
1,2,4-trimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
1,2,5-trimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
1,2-dihydronaphthalene + NADH + O2
cis-(1R,2S)-dihydroxy-1,2,3,4-tetrahydronaphthalene + cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene + NAD+
show the reaction diagram
-
naphthalene dioxygenase activity in cells induced by salicylate
-
?
1,3-benzodithiole + NADH + O2
(R)-1,3-benzodithiole-1-oxide + NAD+
show the reaction diagram
1,4-benzo[d]dithiine + NADH + O2
(S)-1,4-benzo[d]dithiin-1-oxide + NAD+
show the reaction diagram
1,4-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
1,5-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
1-methylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of dioxygenated product, but mainly formation of side-chain monooxygenated product
-
?
2,3-dihydro-1,4-benzo[d]dithiine + NADH + O2
(S)-2,3-dihydrobenzo[d]dithiin-1-oxide + NAD+
show the reaction diagram
2,3-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
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generation of mono- and mostly dioxygenated products
-
?
2,4,6-trimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
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generation of mostly mono- and some dioxygenated products
-
?
2-chlorobiphenyl + NADH + O2
2-chloro-2',3'-dihydrobiphenyl-2',3'-diol + NAD+
show the reaction diagram
2-chlorophenol + NADH + O2
chlorohydroquinone + NAD+
show the reaction diagram
2-methoxynaphthalene + NADH + O2
(1R,2S)-dihydroxy-7-methoxy-1,2-dihydronaphthalene + NAD+
show the reaction diagram
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naphtalene dioxygenase activity in whole cells and activity in E. coli cells expressing naphtalene dioxygenase
minor product (1R,2S)-dihydroxy-6-methoxy-1,2-dihydronaphthalene
?
2-methyl-1,3-benzodithiole + NADH + O2
(1S,2R)-2-methyl-1,3-benzodithiole-1-oxide + (1S,2S)-2-methyl-1,3-benzodithiole-1-oxide + NAD+
show the reaction diagram
2-methylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of two different dioxygenated products
-
?
2-methylquinoline + NADH + H+ + O2
(5R,6S)-2-methyl-5,6-dihydroquinoline-5,6-diol + 2-methylquinolin-8-ol + NAD+
show the reaction diagram
2-methylquinoline + NADH + H+ + O2
5,6-dihydroquinolin-8-ylmethanol + (5R,6S)-8-methyl-5,6-dihydroquinoline-5,6-diol + NAD+
show the reaction diagram
3 benz[a]anthracene + 3 NADH + 3 O2 + 3 H+
benz[a]anthracene-10,11-cis-dihydrodiol + benz[a]anthracene-8,9-cis-dihydrodiol + benz[a]anthracene-1,2-cis-dihydrodiol + 3 NAD+
show the reaction diagram
3,4-dihydro-2H-1lambda4,5-benzodithiepin + NADH + O2
(S)-1,2,3,4-tetrahydro-1lambda4,5-benzodithiepin-1-oxide + NAD+
show the reaction diagram
3-chlorobiphenyl + NADH + O2
3-chloro-2',3'-dihydrobiphenyl-2',3'-diol + NAD+
show the reaction diagram
3-chlorophenol + NADH + O2
chlorohydroquinone + 4-chlorocatechol + NAD+
show the reaction diagram
3-methylisoquinoline + NADH + H+ + O2
cis-3-methyl-7,8-dihydroisoquinoline-7,8-diol + 3-methylisoquinolin-5-ol + NAD+
show the reaction diagram
3-nitrotoluene + NADH + O2
4-nitrobenzyl alcohol + NAD+
show the reaction diagram
4-chlorobiphenyl + NADH + O2
4-chloro-2',3'-dihydrobiphenyl-2',3'-diol + NAD+
show the reaction diagram
4-chlorophenol + NADH + O2
4-chlorocatechol + NAD+
show the reaction diagram
6,7-dihydro-5H-benzocycloheptene + NADH + O2
(1R,2S)-cis-dihydroxybenzocycloheptane + NAD+
show the reaction diagram
-
naphthalene dioxygenase activity in cells induced by salicylate
enantiomeric excess greater than 98%
?
9,10-dihydroanthracene + NADH + O2
cis-(1R,2S)-1,2-dihydroxy-1,2,9,10-tetrahydroanthracene + NAD+
show the reaction diagram
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naphthalene dioxygenase activity in cells induced by salicylate
more than 95% yield, enantiomeric excess greater than 95%, minor product: 9-hydroxy-9,10-dihydroanthracene
?
9,10-dihydrophenanthrene + NADH + O2
cis-(3S,4R)-3,4-dihydroxy-3,4,9,10-tetrahydrophenanthrene + (S)-9-hydroxy-9,10-dihydrophenanthrene
show the reaction diagram
-
naphthalene dioxygenase activity in cells induced by salicylate
70% cis-(3S,4R)-3,4-dihydroxy-3,4,9,10-tetrahydrophenanthrene, enantiomeric excess greater than 95%, 30% (S)-9-hydroxy-9,10-dihydrophenanthrene
?
anthracene + NADH + O2
(1R,2S)-1,2-dihydroanthracene-1,2-diol + (1S,2R)-1,2-dihydroanthracene-1,2-diol + NAD+
show the reaction diagram
benzene + NADH + O2
H2O2 + cis-benzene-1,2-dihydrodiol + NAD+
show the reaction diagram
-
40-50% of O2 is reduced to H2O2 by an uncoupling reaction, only trace amounts of cis-benzene-1,2-dihydrodiol are formed
-
?
benzocyclobutene + NADH + O2
benzocyclobutene-1-ol + benzocyclobutene-1-one
show the reaction diagram
-
naphtalene dioxygenase activity in intact cells
-
?
benz[a]anthracene-1,2-cis-dihydrodiol + NADH + O2
benz[a]anthracene-1,2,10,11-bis-cis-dihydrodiol + NAD+
show the reaction diagram
benz[a]anthracene-10,11-cis-dihydrodiol + NADH + O2
benz[a]anthracene-1,2,10,11-bis-cis-dihydrodiol + NAD+
show the reaction diagram
biphenyl + NADH + O2
? + NAD+
show the reaction diagram
chrysene + NADH + O2
chrysene-3,4-cis-dihydrodiol + chrysene-3,4,9,10-bis-cis-dihydrodiol + NAD+
show the reaction diagram
-
-
-
-
?
dibenzofuran + NADH + O2
(1R,2S)-cis-1,2-dihydroxy-1,2-dihydrodibenzofuran + (3S,4R)-cis-3,4-dihydroxy-3,4-dihydrodibenzofuran + NAD+
show the reaction diagram
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naphthalene dioxygenase activity in cells induced by salicylate
-
-
-
dibenzothiophene + NADH + O2
(1R,2S)-cis-1,2-dihydroxy-dihydrodibenzothiophene + dibenzothiophene sulfoxide
show the reaction diagram
-
naphthalene dioxygenase activity in cells induced by salicylate
84-87% (1R,2S)-cis-1,2-dihydroxy-dihydrodibenzothiophene, enantiomeric excess greater than 95%, 15% dibenzothiophene sulfoxide
?
fluorene + NADH + O2
(3S,4R)-cis-3,4dihydroxy-3,4-dihydrofluorene + 9-fluorenol + NAD+
show the reaction diagram
-
naphthalene dioxygenase activity in cells induced by salicylate
-
-
-
fulvic acid + NADH + H+ + O2
?
show the reaction diagram
humic acid + NADH + H+ + O2
?
show the reaction diagram
indan + NADH + O2
(1S)-indenol + (1S)-indanol + NAD+
show the reaction diagram
-
recombinant enzyme, enantiospecific monooxygenation
20% indenol, 67.1% indanol, minor products: indene, 1-indanone and (1R,2S)-indandiol
?
indan + NADH + O2
1-indanol + NAD+
show the reaction diagram
-
-
-
?
indene + NADH + O2
(1S)-indenol + (1R,2S)-indandiol + NAD+
show the reaction diagram
-
recombinant enzyme
57.9% indenol, 29.6% indandiol
?
indole + ?
2,3-dihydroxy-2,3-dihydroindole + ?
show the reaction diagram
indole + ? + O2
cis-indole-2,3-dihydrodiol
show the reaction diagram
indole + NADH + H+ + O2
?
show the reaction diagram
indole + NADH + O2
(2S,3S)-2,3-dihydro-1H-indole-2,3-diol + NAD+
show the reaction diagram
m-cresol + NADH + O2
methylhydroquinone + 3-methylcatechol + 4-methylcatechol + NAD+
show the reaction diagram
naphthalene + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
naphthalene + NADH + H+ + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
naphthalene + NADH + H+ + O2
1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
naphthalene + NADH + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
o-cresol + NADH + O2
methylhydroquinone + 3-methylcatechol + NAD+
show the reaction diagram
-
-
-
-
?
p-cresol + NADH + O2
4-methylcatechol + NAD+
show the reaction diagram
-
-
-
-
?
phenanthrene + NADH + O2
(1R,2S)-1,2-dihydrophenanthrene-1,2-diol + (1S,2R)-1,2-dihydrophenanthrene-1,2-diol + (3R,4S)-3,4-dihydrophenanthrene-3,4-diol + (3S,4R)-3,4-dihydrophenanthrene-3,4-diol + NAD+
show the reaction diagram
phenanthrene + NADH + O2
? + NAD+
show the reaction diagram
-
-
-
-
?
phenol + NADH + O2
catechol + hydroquinone + NAD+
show the reaction diagram
-
-
-
-
?
quinazoline + NADH + H+ + O2
(5R,6S)-5,6-dihydroquinazoline-5,6-diol + quinazolin-8-ol
show the reaction diagram
quinoline + NADH + H+ + O2
(5R,6S)-5,6-dihydroquinoline-5,6-diol + quinolin-3-ol + NAD+
show the reaction diagram
quinoxaline + NADH + H+ + O2
(5R,6S)-5,6-dihydroquinoxaline-5,6-diol + NAD+
show the reaction diagram
-
-
exclusive product
-
?
styrene + NADH + O2
(R)-1-phenyl-1,2-ethanediol + NAD+
show the reaction diagram
-
recombinant enzyme system
78.6% enantiomeric excess
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no significant activation by the addition of Fe2+, Fe3+, Zn2+, Mg2+ or Cu2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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10 mM, 63% inhibition of ferredoxinNAP reductase
4-chloromercuribenzoate
-
0.0005 mM, 94% inhibition of ferredoxinNAP reductase
H2O2
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0.4 mM, complete inactivation of reduced terminal dioxgenase ISPNAP after 10 min in the absence of ferrous iron
iodoacetate
-
10 mM, 50% inhibition of ferredoxinNAP reductase
N-ethylmaleimide
-
2 mM, 30% inhibition of ferredoxinNAP reductase
NaN3
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40 mM, 46% inhibition of ferredoxinNAP reductase
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ferricyanide
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activation, direct reduction by reductaseNAP in the presence of NADH in vitro
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00042
Biphenyl
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at 30C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0
0.3
indole
-
from indigo formation of whole cells
0.00092 - 2.9
Naphthalene
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.01
Biphenyl
Sphingomonas sp.
-
at 30C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0
1.82
Naphthalene
Sphingomonas sp.
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at 30C in reaction mixtures containing 0.00013 mM ht-PhnI and 0.5 mM NADH in 50 mM potassium phosphate buffer, pH 7.0
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.006
-
activity of terminal dioxygenase ISPNAR
1.37
-
in the presence of partially purified components A and C of the 3 component enzyme system plus FAD
397
-
cytochrome c reduction by ferredoxinNAP reductase
additional information
-
0.000125 mM/min/mg dry cell weight, naphthalene dioxygenase activity in cells grown on succinate in the presence of naphthalene as inducer
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
-
assay at
7.5
-
assay at
additional information
-
ferredoxin reductaseNAP, pI: 6.3
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
Pseudomonas sp. (strain C18)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13500
-
ferredoxin component, gel filtration
34900
-
ferredoxinNAP reductase, native PAGE
37000
-
ferredoxinNAP reductase, gel filtration
37100
-
ferredoxinNAP reductase, deduced from amino acid sequence
45000
-
reductase component, gel filtration
99000
-
gel filtration
155000
-
terminal dioxygenase ISPNAR, gel filtration
158000
-
component B, i.e. oxygenase ISPNAP, gel filtration
200000
-
oxydase component, gel filtration
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
1 * 43000 + 1 * 56000, SDS-PAGE
hexamer
monomer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallized at 8C using the vapor-diffusion method with a reservoir solution of 2 M (NH4)2SO4 and 2-3% dioxane in 50 mM 2-morpholinoethanesulfonic acid, pH 6.0
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hanging drop vapour diffusion method
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modeling of structure with naphthalene bound at the active site. The mononuclear iron is coordinated by amino acid residues His208, His213, and Asp362 and a dioxygen molecule is bound in a side-on fashion. The substrate, naphthalene, is found 2.7 to 3.0 A below the dioxygen molecule and is confined in a small substrate binding site which is surrounded by several hydrophobic residues. The amino acid residues with aromatic side chains, including Phe224, Phe202, Phe352, Trp358, and His295 form a small, but rigid, substrate binding site
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vapor diffusion, equal volume of enzyme solution, 30 mg/ml, and reservoir solution, 2 M ammonium sulfate, 2-3% dioxane in 50 mM Mes, pH 6.0, mixed on a cover slip, deep red crystals, refinement of X-ray structure at 2.25 A resolution
-
diffraction-quality crystals by hangig-drop method, 2.3 A resolution
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hanging drop vapour diffusion method using 2-methyl-2,4-pentanediol as a precipitant
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
room temperature, ferredoxinNAP reductase, t1/2: 8 h
95
-
the enzyme does not completely unfold even at 95C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
10% ethanol and 10% glycerol are required for stability
-
dialysis inactivates rapidly, NaBH4 and dithioerythritol restore activity
-
dilution inactivates rapidly, NaBH4 and dithioerythritol restore activity
-
DTT stabilizes
-
purification of reductase leads to significant loss of flavin cofactor
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2-methoxyethanol
-
stable to
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, ferredoxinNAP reductase, 1 month, minimal loss of activity, prolonged storage leads to precipitation when preparation is heated above 5C
-
0-5C, ferredoxinNAP reductase, 5 days, 30% loss of activity
-
23-25C, buffer, 14 days, loss of about 80% activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography separates the reductase from the two other enzyme components; purification of ferredoxinNAP reductase, ferredoxinNAP and ISPNAP
-
ammonium sulfate, Sephadex G-75, DEAE-cellulose
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Blue Sepharose, Q-Sepharose, Superose 12, purification of component B, the terminal oxygenase ISPNAR
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component B i.e. the terminal oxygenase ISPNAP, DEAE-Sephadex, DEAE-cellulose, octyl-Sepharose, presence of 10% ethanol and 10% glycerol is required to maintain stability
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component B i.e. the terminal oxygenase ISPNAP, DEAE-Sephadex, DEAE-cellulose, octyl-Sepharose, presence of 10% ethanol and 10% glycerol is required to maintain stability; ferredoxinNAP reductase, Blue Sepharose, DEAE-cellulose; purification of ferredoxinNAP reductase, ferredoxinNAP and ISPNAP
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DEAE-cellulose DE52 column chromatography, immobilized metal affinity chromatography and DEAE cellulose column chromatography
-
ferredoxinNAP reductase, Blue Sepharose, DEAE-cellulose
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metal affinity chromatography
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Q-Sepharose FF column chromatography and 1 hydroxyapatite ion-exchange column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli DH5alpha cells
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expressed in Escherichia coli strain K-12
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expressed in Pseudomonas putida
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expression in Escherichia coli
expression of ferredoxinNAP reductase, ferredoxinNAP and ISPNAP in Escherichia coli JM 109
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expression of histidine tagged enzyme in Escherichia coli
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His-tagged, expressed in Escherichia coli
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strain G7, fragment of plasmid NAH7 cloned and expressed in Escherichia coli HB101
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strain NCIB 9816, nahA-gene containing ndoA, ndoB and ndoC is inserted into plasmid pT7-5, transferred to and expressed in Escherichia coli HB101
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D362A
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mutation in alpha subunit of terminal dioxgenase ISPNAP, no activity with naphthalene, biphenyl and phenanthrene
F202L
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, no activity with naphthalene, biphenyl and phenanthrene
F202V
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
F224A
-
6.8% of wild-type activity with substrate naphthalene
F224C
-
9.1% of wild-type activity with substrate naphthalene
F224D
-
85.2% of wild-type activity with substrate naphthalene
F224E
-
94.7% of wild-type activity with substrate naphthalene
F224G
-
2.1% of wild-type activity with substrate naphthalene
F224H
-
92% of wild-type activity with substrate naphthalene
F224I
-
41.9% of wild-type activity with substrate naphthalene
F224K
-
95.3% of wild-type activity with substrate naphthalene
F224L
-
92.58% of wild-type activity with substrate naphthalene
F224M
-
97.5% of wild-type activity with substrate naphthalene
F224N
-
94.7% of wild-type activity with substrate naphthalene
F224P
-
15.1% of wild-type activity with substrate naphthalene
F224Q
-
20.4% of wild-type activity with substrate naphthalene
F224R
-
0.7% of wild-type activity with substrate naphthalene
F224S
-
2% of wild-type activity with substrate naphthalene
F224T
-
4.9% of wild-type activity with substrate naphthalene
F224V
-
13.2% of wild-type activity with substrate naphthalene
F224W
-
91.8% of wild-type activity with substrate naphthalene
F224Y
-
97% of wild-type activity with substrate naphthalene
F352L
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type, major product of biphenyl oxidation: biphenyl-3,4-dihydrodiol instead of biphenyl-2,3-dihydrodiol
M366W
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
N201A
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene as wild-type, very weak activity with biphenyl
N201Q
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
N201S
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
T351N
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
T351R
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
T351S
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
V260A
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
V260L
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
V260N
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
W316A
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type
W358A
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, very low activity with naphthalene and biphenyl
F224A
Pseudomonas sp. NCIB 9816-4
-
6.8% of wild-type activity with substrate naphthalene
-
F224C
Pseudomonas sp. NCIB 9816-4
-
9.1% of wild-type activity with substrate naphthalene
-
F224D
Pseudomonas sp. NCIB 9816-4
-
85.2% of wild-type activity with substrate naphthalene
-
F224S
Pseudomonas sp. NCIB 9816-4
-
2% of wild-type activity with substrate naphthalene
-
F224Y
Pseudomonas sp. NCIB 9816-4
-
97% of wild-type activity with substrate naphthalene
-
F352V
Pseudomonas sp. NCIB 9816-4
-
mutation changes the product regioselectivities for naphthalene, phenanthrene, and biphenyl
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
oxidation of naphthalene to produce optically pure (+)-cis-(1R,2S)-1,2-napthalene dihydrodiol. Increase of productivity of the biotransformation by using resting cells. The biocatalyst is recycled for at least four runs in both suspended and immobilized form. Suspended resting cells retain their activity for at least four runs for 6 h recycle, but the stability is not retained for more than two runs for 12 h recycle. The stability of the 12 h recycle is improved by immobilization