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
1.14.12.12
-
RECOMMENDED NAME
GeneOntology No.
naphthalene 1,2-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
proposed electron transport chain: NADH, ferredoxixinNAP reductase, ferredoxinNAP, terminal oxygenase ISPNAP
-
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
proposed mechanism
-
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
proposed mechanism
Pseudomonas putida 119
-
-
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
proposed electron transport chain: NADH, ferredoxixinNAP reductase, ferredoxinNAP, terminal oxygenase ISPNAP
Pseudomonas sp. NCIB 9816
-
-
naphthalene + NADH + H+ + O2 = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidation
-
naphthalene dioxygenase has a relaxed substrate specificity and can oxidize almost 100 substrates, these include the enantiospecific cis-dihydroxylation of polycyclic aromatic hydrocarbons and the olefin groups of benzocycloalkenes, benzylic hydroxylation, N- and O-dealkylation, sulfoxidation and desaturation reactions
oxidation
Pseudomonas sp. NCIB 9816-4
-
naphthalene dioxygenase has a relaxed substrate specificity and can oxidize almost 100 substrates, these include the enantiospecific cis-dihydroxylation of polycyclic aromatic hydrocarbons and the olefin groups of benzocycloalkenes, benzylic hydroxylation, N- and O-dealkylation, sulfoxidation and desaturation reactions
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Ethylbenzene degradation
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
Naphthalene degradation
-
naphthalene degradation (aerobic)
-
Polycyclic aromatic hydrocarbon degradation
-
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+.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
naphthalene 1,2-dioxygenase
-
-
naphthalene 1,2-dioxygenase
Pseudomonas putida G7
-
-
-
naphthalene dioxygenase
-
-
-
-
naphthalene dioxygenase
-
-
naphthalene dioxygenase
Escherichia coli JM109(DE3)
-
-
-
naphthalene dioxygenase
-
-
naphthalene dioxygenase
P0A110 and P0A112
-
naphthalene dioxygenase
Pseudomonas putida 8859
-
-
-
naphthalene dioxygenase
Pseudomonas putida G7
P0A110 and P0A112
-
-
naphthalene dioxygenase
-
-
naphthalene dioxygenase
Pseudomonas sp. HOB1, Pseudomonas sp. NCIB 9816-4
-
-
-
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)
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)
-
naphthalene oxygenase
-
-
-
-
NDO
Escherichia coli JM109(DE3)
-
-
-
NDO
Pseudomonas putida 8859
-
-
-
NDO
Pseudomonas sp. NCIB 9816-4
-
;
-
oxygenase, naphthalene di-
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9074-04-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
grown on naphthalene as sole carbon source
-
-
Manually annotated by BRENDA team
strain JM109(DE3)
-
-
Manually annotated by BRENDA team
Escherichia coli JM109(DE3)
strain JM109(DE3)
-
-
Manually annotated by BRENDA team
strain NCIMB 40531
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens NCIMB 40531
strain NCIMB 40531
-
-
Manually annotated by BRENDA team
large alpha subunit and small beta subunit
P0A110 and P0A112
UniProt
Manually annotated by BRENDA team
strain 119, unable to grow on naphthalene; wild-type grown on naphthalene
-
-
Manually annotated by BRENDA team
Pseudomonas putida 119
strain 119, unable to grow on naphthalene
-
-
Manually annotated by BRENDA team
Pseudomonas putida 8859
8859
-
-
Manually annotated by BRENDA team
Pseudomonas putida G7
G7
-
-
Manually annotated by BRENDA team
Pseudomonas putida G7
large alpha subunit and small beta subunit
P0A110 and P0A112
UniProt
Manually annotated by BRENDA team
Pseudomonas putida G7
strain G7
-
-
Manually annotated by BRENDA team
NCIB 9816-4
-
-
Manually annotated by BRENDA team
NCIB 9816-4. Enzyme is recombinantly expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
strain HKT554
-
-
Manually annotated by BRENDA team
strain HOB1
-
-
Manually annotated by BRENDA team
strain NCIB 9816
-
-
Manually annotated by BRENDA team
strain NCIB 9816-4
-
-
Manually annotated by BRENDA team
strain NCIB 9816/11, naphthalene dihydrogenase mutant derived from strain NCIB 9816-4 oxidizing naphthalene to cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene
-
-
Manually annotated by BRENDA team
strain HKT554
-
-
Manually annotated by BRENDA team
Pseudomonas sp. HOB1
strain HOB1
-
-
Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9616-4
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9816
strain NCIB 9816
-
-
Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9816-4
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9816-4
strain NCIB 9816-4
-
-
Manually annotated by BRENDA team
strain NCIMB12038
-
-
Manually annotated by BRENDA team
strain CHY-1
-
-
Manually annotated by BRENDA team
strain CHY-1
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
no substrate for wild-type, substrate for mutants F224C, F224K, F224Q, F224Y
product has an oxygen atom directly attached to the aromatic ring. Upon completion of the dihydroxylation, the oxygen atom on the C-ring is believed to expel the 7-OH group through donation of its lone pair electrons and the charged intermediate. Once formed this readily re-aromatizes to provide 8-hydroxyflavanones or 8-hydroxyisoflavanones
-
?
(3S)-isoflavanone + NADH + H+ + O2
8-hydroxy-(3S)-isoflavanone + NAD+ + ?
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
no substrate for wild-type, substrate for mutants F224K, F224Y
product has an oxygen atom directly attached to the aromatic ring. Upon completion of the dihydroxylation, the oxygen atom on the C-ring is believed to expel the 7-OH group through donation of its lone pair electrons and the charged intermediate. Once formed this readily re-aromatizes to provide 8-hydroxyflavanones or 8-hydroxyisoflavanones
-
?
(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
-
-
generation of mono- and dioxygenated products
-
?
1,2,5-trimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of mostly mono- and some dioxygenated products
-
?
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
Pseudomonas putida, Pseudomonas putida 8859
-
-
-
-
?
1,4-benzo[d]dithiine + NADH + O2
(S)-1,4-benzo[d]dithiin-1-oxide + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida 8859
-
-
-
-
?
1,4-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of monooxygenated products
-
?
1,5-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of monooxygenated products
-
?
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
Pseudomonas putida, Pseudomonas putida 8859
-
-
-
-
?
2,3-dimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
generation of mono- and mostly dioxygenated products
-
?
2,4,6-trimethylnaphthalene + NADH + H+ + O2
? + NAD+
show the reaction diagram
-
-
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
Pseudomonas putida, Pseudomonas putida G7
-
recombinant histidine-tagged terminal dioxygenase
-
?
2-chlorophenol + NADH + O2
chlorohydroquinone + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
2-methoxynaphthalene + NADH + O2
(1R,2S)-dihydroxy-7-methoxy-1,2-dihydronaphthalene + NAD+
show the reaction diagram
-
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
Pseudomonas putida, Pseudomonas putida 8859
-
-
cis and trans product
-
?
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
Pseudomonas sp., Pseudomonas sp. NCIB 9616-4
-
-
99% total conversion, giving 92% (5R,6S)-2-methyl-5,6-dihydroquinoline-5,6-diol and 8% 2-methylquinolin-8-ol
-
?
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
Pseudomonas sp., Pseudomonas sp. NCIB 9616-4
-
-
78% total conversion, giving 72% 5,6-dihydroquinolin-8-ylmethanol + 28% (5R,6S)-8-methyl-5,6-dihydroquinoline-5,6-diol
-
?
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
Pseudomonas putida, Pseudomonas putida 8859
-
-
-
-
?
3-chlorobiphenyl + NADH + O2
3-chloro-2',3'-dihydrobiphenyl-2',3'-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
recombinant histidine-tagged terminal dioxygenase
-
?
3-chlorophenol + NADH + O2
chlorohydroquinone + 4-chlorocatechol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
3-methylisoquinoline + NADH + H+ + O2
cis-3-methyl-7,8-dihydroisoquinoline-7,8-diol + 3-methylisoquinolin-5-ol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9616-4
-
-
99% total conversion, giving 73% cis-3-methyl-7,8-dihydroisoquinoline-7,8-diol + 27% 3-methylisoquinolin-5-ol
-
?
3-nitrotoluene + NADH + O2
4-nitrobenzyl alcohol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
4-chlorobiphenyl + NADH + O2
4-chloro-2',3'-dihydrobiphenyl-2',3'-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
recombinant histidine-tagged terminal dioxygenase
-
?
4-chlorophenol + NADH + O2
4-chlorocatechol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
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
-
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
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
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
-
-
-
-
?
biphenyl + NADH + O2
? + NAD+
show the reaction diagram
-
-
-
-
?
biphenyl + NADH + O2
? + NAD+
show the reaction diagram
Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
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
-
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
Pseudomonas putida, Pseudomonas putida G7
-
-
-
-
?
humic acid + NADH + H+ + O2
?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
-
-
-
?
indan + NADH + O2
1-indanol + NAD+
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
?
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
Pseudomonas sp., Pseudomonas sp. HOB1
-
-
-
-
?
indole + ? + O2
cis-indole-2,3-dihydrodiol
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
reaction of recombinant E. coli transformed with Pseudomonas sp. DNA
-
?
indole + NADH + H+ + O2
?
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
-
-
-
?
indole + NADH + O2
(2S,3S)-2,3-dihydro-1H-indole-2,3-diol + NAD+
show the reaction diagram
-
-
-
-
?
indole + NADH + O2
(2S,3S)-2,3-dihydro-1H-indole-2,3-diol + NAD+
show the reaction diagram
-
-
-
-
?
indole + NADH + O2
(2S,3S)-2,3-dihydro-1H-indole-2,3-diol + NAD+
show the reaction diagram
Escherichia coli JM109(DE3)
-
-
-
-
?
indole + NADH + O2
(2S,3S)-2,3-dihydro-1H-indole-2,3-diol + 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 + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
-
-
-
?
naphthalene + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
-
-
only cis-isomer by bacteria, product rapidly autooxidizes to 1,2-naphthoquinone
?
naphthalene + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
-
50% activity with NADH compared to NADPH
-
-
-
naphthalene + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
Pseudomonas putida 119
-
-
only cis-isomer by bacteria, product rapidly autooxidizes to 1,2-naphthoquinone
?
naphthalene + NAD(P)H + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(P)+
show the reaction diagram
Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
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
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
-
exclusive product
-
?
naphthalene + NADH + H+ + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
the catalytic ferrous site is primed for the O2 reaction when substrate is bound in the active site in the presence of the reduced Rieske site. These structural changes ensure that two electrons and the substrate are present before the binding and activation of O2, which avoids the uncontrolled formation and release of reactive oxygen species
-
-
?
naphthalene + NADH + H+ + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
-
exclusive product
-
?
naphthalene + NADH + H+ + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
naphthalene + NADH + H+ + O2
1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida, Pseudomonas putida G7
-
-
-
-
?
naphthalene + NADH + O2
cis-(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
-
-
-
?
naphthalene + NADH + O2
cis-(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
-
-
-
?
naphthalene + NADH + O2
cis-(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
-
-
-
?
naphthalene + NADH + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
3 component enzyme system consisting of ferredoxinNAP reductase, ferredoxinNAP and oxygenase ISPNAP, ferredoxinNAP reductase reduces: 2,6-dichlorophenolindophenol, ferricyanide, nitro blue tetrazolium and cytochrome c, in the presence of FAD ferredoxinNAP reductase transfers electrons to ferredoxin
-
?
naphthalene + NADH + O2
cis-(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
?
naphthalene + NADH + O2
(1R,2S)-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
(1R,2S)-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
(1R,2S)-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
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
three-component dioxygenase, uses two proteins containing three redox centers to transfer electrons to the terminal oxygenase
-
-
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
genes coding for naphthalene dioxygenase: ndoA, ndoB and ndoC
-
-
naphthalene + NADH + O2
(1R,2S)-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
Pseudomonas putida 119
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
naphthalene + NADH + O2
(1R,2S)-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
Pseudomonas sp. NCIB 9816
-
-
-
-
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
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
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
Pseudomonas sp., Pseudomonas sp. NCIB 9616-4
-
-
99% total conversion, givng 56% (5R,6S)-5,6-dihydroquinazoline-5,6-diol + 43% quinazolin-8-ol
-
?
quinoline + NADH + H+ + O2
(5R,6S)-5,6-dihydroquinoline-5,6-diol + quinolin-3-ol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9616-4
-
-
66% total conversion, giving 55% (5R,6S)-5,6-dihydroquinoline-5,6-diol and 35% quinolin-3-ol
-
?
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
?
m-cresol + NADH + O2
methylhydroquinone + 3-methylcatechol + 4-methylcatechol + NAD+
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816-4
-
-
-
-
?
additional information
?
-
-
the enzyme is responsible for the initial catabolic reactions onto naphthothiophenes and dibenzothiophene
-
-
-
additional information
?
-
-
biotransformation of 2-pyridone- and 2-quinolone-derived compounds by recombinant whole-cells of Escherichia coli JM109(DE3)(pDTG141) expressing the naphthalene-dioxygenase system from Pseudomonas sp. NCIB 9816-4, using a series of N- and C-substituted derivatives. Among them, only the N-methyl substituted compounds are good substrates for a regio- and stereoselective dihydroxylation reaction leading to cis-dihydroxydihydro pyridone derivatives
-
-
-
additional information
?
-
-
the enzyme is responsible for the initial catabolic reactions onto naphthothiophenes and dibenzothiophene
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
-
-
-
-
naphthalene + NADH + O2
(1R,2S)-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
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
three-component dioxygenase, uses two proteins containing three redox centers to transfer electrons to the terminal oxygenase
-
-
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
-
genes coding for naphthalene dioxygenase: ndoA, ndoB and ndoC
-
-
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas putida 119
-
initial oxidative reaction in bacterial naphthalene catabolism
-
?
naphthalene + NADH + O2
(1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD+
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
-
additional information
?
-
-
the enzyme is responsible for the initial catabolic reactions onto naphthothiophenes and dibenzothiophene
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FAD
-
1 mol FAD/mol enzyme in flavin-reconstituted protein; addition enhances ferredoxinNAP reductase activity with all in vitro electron-acceptors, e.g. cytochrome c, 2,6-dichlorophenolindophenol, Nitroblue tetrazolium and ferricyanide; component A, ferredoxinNAP reductase is a flavoprotein, enzyme can use both FAD and FMN but exhibits slightly higher affinity for FAD
FAD
-
component A, ferredoxinNAP reductase is a flavoprotein, enzyme can use both FAD and FMN but exhibits slightly higher affinity for FAD
FAD
-
stimulates cytochrome c reduction by ferredoxinNAP reductase
FAD
-
stimulates activity of terminal oxygenase ISPNAR
FMN
-
requirement, ferredoxinNAP reductase is a flavoprotein, enzyme can use both FAD and FMN but exhibits slightly higher affinity for FAD, addition stimulates dioxygenase activity by 53% of FAD-stimulation
FMN
-
stimulates cytochrome c reduction by ferredoxinNAP reductase
NADH
-
requirement, the oxygenase accepts two electrons from NADH, the reduction requires component A and C as mediators
NADPH
-
can replace NADH with 39%; ferredoxinNAP reductase
NADPH
-
less than 50% of the activity with NADH
FMN
-
stimulates activity of terminal oxygenase ISPNAR
additional information
-
spinach ferredoxin cannot replace ferredoxinNAP
-
additional information
-
cytochrome P-450 is no cofactor
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
component A, i.e. NADH-ferredoxinNAP reductase, of the multienzyme system is an iron-containing flavoprotein containing 1.8 g-atoms Fe2+ and 2 g-atoms sulfur
Fe2+
-
component B, the terminal oxygenase ISPNAP is an iron-sulfur protein, oxidized ISPNAP binds naphthalene without conformational changes that affect its FeS-chromophores, ISPNAP contains 6 g-atom Fe2+ and 4 g-atom acid-labile sulfur per mol enzyme, the enzyme complex is not stimulated by exogenous Fe2+
Fe2+
-
non-heme iron protein
Fe2+
-
tightly bound Fe2+
Fe2+
-
terminal dioxygenase ISPNAR may be a Rieske-type iron-sulfur protein containing 2.4 g-atoms of iron and 2.1 g-atoms of sulfur per alpha,beta subunit
Fe2+
-
alpha subunit of terminal oxygenase ISPNAP contains a Rieske [2Fe-2S] center in one domain and a mono-nuclear iron in the catalytic domain
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
Iron
-
a molecular oxygen species is bound to the mononuclear iron in a side-on fashion
Iron
-
in the presence of the aromatic substrate indole, nitric oxide is bound end-on to the active-site mononuclear iron of NDO
Iron
-
the catalytic ferrous site is primed for the O2 reaction when substrate is bound in the active site in the presence of the reduced Rieske site. These structural changes ensure that two electrons and the substrate are present before the binding and activation of O2, which avoids the uncontrolled formation and release of reactive oxygen species
additional information
-
no significant activation by the addition of Fe2+, Fe3+, Zn2+, Mg2+ or Cu2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
10 mM, 63% inhibition of ferredoxinNAP reductase
4-chloromercuribenzoate
-
0.0005 mM, 94% inhibition of ferredoxinNAP reductase
H2O2
-
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
-
40 mM, 46% inhibition of ferredoxinNAP reductase
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ferricyanide
-
activation, direct reduction by reductaseNAP in the presence of NADH in vitro
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00042
-
Biphenyl
-
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
-
Naphthalene
-
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
2.9
-
Naphthalene
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.01
-
Biphenyl
-
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
-
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]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
-
-
assay at
7.5
-
-
assay at
additional information
-
-
ferredoxin reductaseNAP, pI: 6.3
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Pseudomonas sp. NCIB 9816
-
;
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
three component enzyme system consisting of: component A, i.e. ferredoxinNAP reductase, component B, i.e. terminal oxygenase ISPNAP, and component C, i.e. ferredoxinNAP
additional information
-
-
three component enzyme system consisting of: component A, i.e. ferredoxinNAP reductase, component B, i.e. terminal oxygenase ISPNAP, and component C, i.e. ferredoxinNAP
additional information
-
-
three component enzyme system consisting of: component A, i.e. ferredoxinNAP reductase, component B, i.e. terminal oxygenase ISPNAP, and component C, i.e. ferredoxinNAP
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 53000 + x * 25000, histidine-tagged recombinant enzyme, SDS-PAGE
?
Pseudomonas putida G7
-
x * 53000 + x * 25000, histidine-tagged recombinant enzyme, SDS-PAGE
-
dimer
-
1 * 43000 + 1 * 56000, SDS-PAGE
hexamer
-
3 * 52000, 3 * 20000, alpha3beta3 hexamer, oxygenase component, SDS-PAGE
hexamer
-
x-ray crystallography
hexamer
-
x-ray crystallography
-
hexamer
-
3 * 52000, 3 * 20000, alpha3beta3 hexamer, oxygenase component, SDS-PAGE
-
monomer
-
1 * 36000, ferredoxinNAP reductase, SDS-PAGE
monomer
-
1 * 13500, ferredoxin component, gel filtration
monomer
Pseudomonas sp. NCIB 9816
-
1 * 36000, ferredoxinNAP reductase, SDS-PAGE
-
tetramer
-
alpha2beta2 2 * 55000 + 2 * 20000, terminal oxygenase ISPNAP, SDS-PAGE
tetramer
-
alpha2beta2, 2 * 55000 + 2 * 23000, SDS-PAGE
tetramer
Pseudomonas sp. NCIB 9816
-
alpha2beta2 2 * 55000 + 2 * 20000, terminal oxygenase ISPNAP, SDS-PAGE
-
tetramer
-
alpha2beta2, 2 * 55000 + 2 * 23000, SDS-PAGE
-
monomer
-
1 * 13500, ferredoxin component, gel filtration
-
additional information
-
X-ray structure data from Pseudomonas sp. 9861-4 suggest an alpha3,beta3 hexameric structure
additional information
-
X-ray structure data from Pseudomonas sp. 9861-4 suggest an alpha3,beta3 hexameric structure
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
hanging drop vapour diffusion method
-
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
-
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
-
hanging drop vapour diffusion method using 2-methyl-2,4-pentanediol as a precipitant
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
room temperature, ferredoxinNAP reductase, t1/2: 8 h
95
-
-
the enzyme does not completely unfold even at 95C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
2-methoxyethanol
-
stable to
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
23-25C, buffer, 14 days, loss of about 80% activity
-
-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
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate, Sephadex G-75, DEAE-cellulose
-
metal affinity chromatography
-
affinity chromatography separates the reductase from the two other enzyme components; purification of ferredoxinNAP reductase, ferredoxinNAP and ISPNAP
-
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
-
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
-
ferredoxinNAP reductase, Blue Sepharose, DEAE-cellulose
-
Q-Sepharose FF column chromatography and 1 hydroxyapatite ion-exchange column chromatography
-
Blue Sepharose, Q-Sepharose, Superose 12, purification of component B, the terminal oxygenase ISPNAR
-
DEAE-cellulose DE52 column chromatography, immobilized metal affinity chromatography and DEAE cellulose column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of histidine tagged enzyme in Escherichia coli
-
His-tagged, expressed in Escherichia coli
-
strain G7, fragment of plasmid NAH7 cloned and expressed in Escherichia coli HB101
-
expressed in Escherichia coli DH5alpha cells
-
expressed in Escherichia coli strain K-12
-
expression in Escherichia coli
-
expression of ferredoxinNAP reductase, ferredoxinNAP and ISPNAP in Escherichia coli JM 109
-
strain NCIB 9816, nahA-gene containing ndoA, ndoB and ndoC is inserted into plasmid pT7-5, transferred to and expressed in Escherichia coli HB101
-
expression in Escherichia coli
-
expressed in Pseudomonas putida
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D362A
-
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
F352V
-
mutation in alpha subunit of terminal dioxgenase ISPNAP, similar activity with naphthalene, biphenyl and phenanthrene as wild-type, product of biphenyl oxidation: 96% biphenyl-3,4-dihydrodiol instead of biphenyl-2,3-dihydrodiol
F352V
-
mutation changes the product regioselectivities for naphthalene, phenanthrene, and biphenyl
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 ACCESSION NO.
COMMENTARY
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