Information on EC 1.14.13.215 - protoasukamycin 4-monooxygenase

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The expected taxonomic range for this enzyme is: Streptomyces nodosus

EC NUMBER
COMMENTARY hide
1.14.13.215
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RECOMMENDED NAME
GeneOntology No.
protoasukamycin 4-monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
protoasukamycin + NADH + H+ + O2 = 4-hydroxyprotoasukamycin + NAD+ + H2O
show the reaction diagram
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SYSTEMATIC NAME
IUBMB Comments
protoasukamycin,NADH:oxygen oxidoreductase (4-hydroxylating)
The enzyme, characterized from the bacterium Streptomyces nodosus subsp. asukaensis, is involved in the biosynthesis of the antibiotic asukamycin. Requires a flavin cofactor, with no preference among FMN, FAD or riboflavin. When flavin concentration is low, activity is enhanced by the presence of the NADH-dependent flavin-reductase AsuE2.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
subsp. asukaensis
UniProt
Manually annotated by BRENDA team
subsp. asukaensis
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
protoasukamycin + NADH + H+ + O2
4-hydroxyprotoasukamycin + NAD+ + H2O
show the reaction diagram
protoasukamycin + NADPH + H+ + O2
4-hydroxyprotoasukamycin + NADP+ + H2O
show the reaction diagram
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
protoasukamycin + NADH + H+ + O2
4-hydroxyprotoasukamycin + NAD+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FMN
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enzyme AsuE2 contains a trace amount of FMN at a molar ratio of 1:2000 (FMN:AsuE2) upon purification from Escherichia coli
riboflavin
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additional information
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enzyme AsuE1 is a flavin-dependent protoasukamycin hydroxylase, that displays no significant preference toward flavin cofactors, because FAD, FMN, and riboflavin are utilized efficiently in the catalysis
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
protein AsuE2
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0123
NADH
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
1.84
NADPH
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
0.0213
protoasukamycin
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.113
NADH
Streptomyces nodosus
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
0.324
NADPH
Streptomyces nodosus
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
16.67
protoasukamycin
Streptomyces nodosus
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in presence of 0.01 mM FAD, recombinant enzyme, pH 7.0, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chomatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of asukamycin biosynthetic gene cluster, including gene asuE1 encoding the enzyme, recombinant expression in Escherichia coli strain BW25113/pIJ790, heterologous expression of the complete gene cluster in the asukamycin-nonproducing Streptomyces lividans strain K4-114
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gene asuE1, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
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