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Information on EC 1.14.13.180 - aklavinone 12-hydroxylase and Organism(s) Streptomyces peucetius and UniProt Accession P32009

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IUBMB Comments
The enzymes from the Gram-positive bacteria Streptomyces peucetius and Streptomyces purpurascens participate in the biosynthesis of daunorubicin, doxorubicin and rhodomycins. The enzyme from Streptomyces purpurascens is an FAD monooxygenase.
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Streptomyces peucetius
UNIPROT: P32009
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The taxonomic range for the selected organisms is: Streptomyces peucetius
The enzyme appears in selected viruses and cellular organisms
Synonyms
aklavinone 12-hydroxylase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aklavinone 12-hydroxylase
-
SYSTEMATIC NAME
IUBMB Comments
aklavinone,NADPH:oxygen oxidoreductase (12-hydroxylating)
The enzymes from the Gram-positive bacteria Streptomyces peucetius and Streptomyces purpurascens participate in the biosynthesis of daunorubicin, doxorubicin and rhodomycins. The enzyme from Streptomyces purpurascens is an FAD monooxygenase.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
aklavinone + NADPH + H+ + O2
epsilon-rhodomycinone + NADP+ + H2O
show the reaction diagram
C-glucosyl flavokermesic acid + NADPH + H+ + O2
carminic acid + NADP+ + H2O
show the reaction diagram
-
-
-
?
flavokermesic acid + NADPH + H+ + O2
kermesic acid + NADP+ + H2O
show the reaction diagram
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
aklavinone + NADPH + H+ + O2
epsilon-rhodomycinone + NADP+ + H2O
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
the DnrF enzyme contains two motifs that are common to a number of FAD- and NADPH-dependent enzymes. The enzyme catalyses the aklavinone 11-hydroxylation in a NADPH-dependent reaction
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0433 - 0.0568
flavokermesic acid
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
DNRF_STRPE
489
0
52317
Swiss-Prot
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
homology modeling of structure using RdmE from Streptomyces purpurascens, PDB ID 3IHG
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
M56A
2fold increase in synthesis of kermesic acid
P217K
2.2fold increase in synthesis of kermesic acid
P217K/M56A
significant decrease in synthesis of kermesic acid
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
heterologous expression of dnrF in Streptomyces liwidans TK23 and in Escherichia coli. Since the fusion protein does not show any hydroxylase activity, the dnrF gene is cloned into the pT7-7 vector that allows the expression of an unfused DnrF protein
expression in doxorubicin-sensitive Streptomyces galilaeus ATCC 31133. Transformation of Streptomyces galilaeus ATCC 31133 with plasmid pMC213 containing the aklavinone 11-hydroxylase gene (dnrF) results in the production of many red pigments. A new metabolite is purified, and the position of the newly introduced hydroxyl group is determined. The result indicates that the aklavinone 11-hydroxylase gene is stably expressed in Streptomyces galilaeus ATCC 31133 and that it gives rise to a hybrid aclacinomycin A which shows highly specific in vitro cytotoxicity against leukemia and melanoma cell lines
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
biosynthesis of carminic acid from glucose in engineered Escherichia coli. Expression of an optimized type II polyketide synthase machinery from Photorhabdus luminescens enables a high-level production of flavokermesic acid upon coexpression of the cyclases ZhuI and ZhuJ from Streptomyces sp. R1128. Mutants of aklavinone 12-hydroxylase (DnrF) from Streptomyces peucetius and C-glucosyltransferase (GtCGT) from Gentiana triflora with 2-5fold enhanced conversion efficiencies can perform hydroxylation and C-glucosylation of flavokermesic acid, respectively, resulting in 0.63 mg/l of carminic acid production from glucose
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hwang, C.K.; Kim, H.S.; Hong, Y.S.; Kim, Y.H.; Hong, S.K.; Kim, S.J.; Lee, J.J.
Expression of Streptomyces peucetius genes for doxorubicin resistance and aklavinone 11-hydroxylase in Streptomyces galilaeus ATCC 31133 and production of a hybrid aclacinomycin
Antimicrob. Agents Chemother.
39
1616-1060
1995
Streptomyces peucetius
Manually annotated by BRENDA team
Hong, Y.S.; Hwang, C.K., Hong, S.K.; Kim, Y.H.; Lee, J.J.
Molecular cloning and characterization of the aklavinone 11-hydroxylase gene of Streptomyces peucetius subsp. caesius ATCC 27952
J. Bacteriol.
176
7096-7101
1994
Streptomyces peucetius (P72495), Streptomyces peucetius caesius ATCC 27952 (P72495)
Manually annotated by BRENDA team
Filippini, S.; Solinas, M.M.; Breme, U.; Schlter, M.B.; Gabellini, D.; Biamonti, G.; Colombo, A.L.; Garofano, L.
Streptomyces peucetius daunorubicin biosynthesis gene, dnrF: sequence and heterologous expression
Microbiology
141
1007-1016
1995
Streptomyces peucetius (P32009), Streptomyces peucetius ATCC 29050 (P32009)
Manually annotated by BRENDA team
Yang, D.; Jang, W.D.; Lee, S.Y.
Production of carminic acid by metabolically engineered Escherichia coli
J. Am. Chem. Soc.
143
5364-5377
2021
Streptomyces peucetius (P32009), Streptomyces peucetius
Manually annotated by BRENDA team