DOXA_STRPE
ID DOXA_STRPE Reviewed; 415 AA.
AC Q9ZAU3;
DT 19-MAR-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1999, sequence version 1.
DT 03-AUG-2022, entry version 78.
DE RecName: Full=Cytochrome P-450 monooxygenase DoxA;
DE EC=1.14.13.181 {ECO:0000269|PubMed:9864344};
DE AltName: Full=13-deoxycarminomycin C-13 hydroxylase;
DE AltName: Full=13-deoxydaunorubicin C-13 hydroxylase;
DE AltName: Full=13-dihydrocarminomycin C-13 hydroxylase;
DE AltName: Full=13-dihydrodaunorubicin C-13 hydroxylase;
GN Name=doxA;
OS Streptomyces peucetius.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1950;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=ATCC 29050 / DSM 40754 / JCM 9920 / NBRC 100596 / NCIMB 10972;
RX PubMed=9864344; DOI=10.1128/jb.181.1.305-318.1999;
RA Lomovskaya N., Otten S.L., Doi-Katayama Y., Fonstein L., Liu X.-C.,
RA Takatsu T., Inventi A., Filippi S., Torti F., Colombo A.L.,
RA Hutchinson C.R.;
RT "Doxorubicin overproduction in Streptomyces peucetius: cloning and
RT characterization of the dnrU ketoreductase and dnrV genes and the doxA
RT cytochrome P-450 hydroxylase gene.";
RL J. Bacteriol. 181:305-318(1999).
CC -!- FUNCTION: Involved in the biosynthesis of the anthracyclines
CC carminomycin and daunorubicin (daunomycin) which are aromatic
CC polyketide antibiotics that exhibit high cytotoxicity and are widely
CC applied in the chemotherapy of a variety of cancers. In vivo, DoxA
CC catalyzes the C-13 hydroxylation of 13-deoxycarminomycin and 13-
CC deoxydaunorubicin to yield 13-dihydrocarminomycin and 13-
CC dihydrodaunorubicin, respectively, as well as the oxidation of these
CC 13-dihydro-anthracyclines to their respective 13-keto forms,
CC carminomycin and daunorubicin. In vitro, it also catalyzes the C-14
CC hydroxylation of daunorubicin to form doxorubicin (adriamycin),
CC although this strain is not a doxorubicin producer. It can only use
CC NADP. DoxA acts jointly with DnrV. {ECO:0000269|PubMed:9864344}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=13-deoxydaunorubicin + H(+) + NADPH + O2 = 13-
CC dihydrodaunorubicin + H2O + NADP(+); Xref=Rhea:RHEA:37851,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:75296,
CC ChEBI:CHEBI:75297; EC=1.14.13.181;
CC Evidence={ECO:0000269|PubMed:9864344};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=13-dihydrodaunorubicin + H(+) + NADPH + O2 = daunorubicin + 2
CC H2O + NADP(+); Xref=Rhea:RHEA:37847, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:64677, ChEBI:CHEBI:75296;
CC EC=1.14.13.181; Evidence={ECO:0000269|PubMed:9864344};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=13-deoxycarminomycin + H(+) + NADPH + O2 = 13-
CC dihydrocarminomycin + H2O + NADP(+); Xref=Rhea:RHEA:56360,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:140329,
CC ChEBI:CHEBI:140330; Evidence={ECO:0000269|PubMed:9864344};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=13-dihydrocarminomycin + H(+) + NADPH + O2 = carminomycin + 2
CC H2O + NADP(+); Xref=Rhea:RHEA:56364, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:75730, ChEBI:CHEBI:140330;
CC Evidence={ECO:0000269|PubMed:9864344};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- PATHWAY: Antibiotic biosynthesis; daunorubicin biosynthesis.
CC -!- PATHWAY: Antibiotic biosynthesis; carminomycin biosynthesis.
CC -!- SUBUNIT: Monomer. {ECO:0000250}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC -!- MISCELLANEOUS: S.peucetius ATCC 29050 produces significant quantities
CC of doxorubicin only when it grows in a highly-buffered doxorubicin
CC production medium. S.peucetius subsp. caesius ATCC 27952, a mutant
CC strain derived from S.peucetius ATCC 29050, is the only organism
CC reported to produce doxorubicin in vivo (PubMed:9864344).
CC {ECO:0000305|PubMed:9864344}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; U77891; AAD04715.1; -; Genomic_DNA.
DR AlphaFoldDB; Q9ZAU3; -.
DR SMR; Q9ZAU3; -.
DR KEGG; ag:AAD04715; -.
DR BioCyc; MetaCyc:MON-18171; -.
DR BRENDA; 1.14.13.181; 6073.
DR UniPathway; UPA00054; -.
DR UniPathway; UPA01040; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0016709; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR GO; GO:1901771; P:daunorubicin biosynthetic process; IDA:UniProtKB.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR002397; Cyt_P450_B.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00359; BP450.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Antibiotic biosynthesis; Cytoplasm; Heme; Iron; Metal-binding;
KW Monooxygenase; NADP; Oxidoreductase.
FT CHAIN 1..415
FT /note="Cytochrome P-450 monooxygenase DoxA"
FT /id="PRO_0000425684"
FT BINDING 362
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250"
SQ SEQUENCE 415 AA; 45095 MW; CCF873824BEB6CCF CRC64;
MAVDPFACPM MTMQRKPEVH DAFREAGPVV EVNAPAGGPA WVITDDALAR EVLADPRFVK
DPDLAPAAWR GVDDGLDIPV PELRPFTLIA VDGEAHRRLR RIHAPAFNPR RLAERTDRIA
AIAGRLLTEL ADASGRSGKP AELIGGFAYH FPLLVICELL GVPVTDPAMA REAVSVLKAL
GLGGPQSGGG DGTDPAGGVP DTSALESLLL EAVHSARRND TPTMTRVLYE RAQAEFGSVS
DDQLVYMITG LIFAGHDTTG SFLGFLLAEV LAGRLAADAD EDAVSRFVEE ALRYHPPVPY
TLWRFAATEV TIGGVRLPRG APVLVDIEGT NTDGRHHDAP HAFHPDRPSW RRLTFGDGPH
YCIGEQLAQL ESRTMIGVLR SRFPEARLAV PYDELRWCRK GAQTARLTEL PVWLR
