ID   C82D6_SINHE             Reviewed;         519 AA.
AC   A0A0N7F297;
DT   10-FEB-2021, integrated into UniProtKB/Swiss-Prot.
DT   20-JAN-2016, sequence version 1.
DT   03-AUG-2022, entry version 18.
DE   RecName: Full=Demethylepipodophyllotoxin synthase {ECO:0000303|PubMed:26359402};
DE            EC=1.14.14.132 {ECO:0000269|PubMed:26359402};
DE   AltName: Full=Cytochrome P450 family 82 subfamily D polypeptide 61 {ECO:0000303|PubMed:26359402};
GN   Name=CYP82D61 {ECO:0000303|PubMed:26359402};
OS   Sinopodophyllum hexandrum (Himalayan may apple) (Podophyllum hexandrum).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; Ranunculales; Berberidaceae; Podophylloideae;
OC   Sinopodophyllum.
OX   NCBI_TaxID=93608;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY,
RP   TISSUE SPECIFICITY, AND PATHWAY.
RX   PubMed=26359402; DOI=10.1126/science.aac7202;
RA   Lau W., Sattely E.S.;
RT   "Six enzymes from mayapple that complete the biosynthetic pathway to the
RT   etoposide aglycone.";
RL   Science 349:1224-1228(2015).
CC   -!- FUNCTION: Cytochrome P450 involved in the biosynthesis of etoposide, a
CC       chemotherapeutic compound of the topoisomerase inhibitor family
CC       (PubMed:26359402). Catalyzes the hydroxylation of deoxypodophyllotoxin
CC       to form epipodophyllotoxin (PubMed:26359402).
CC       {ECO:0000269|PubMed:26359402}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(-)-4'-desmethyl-deoxypodophyllotoxin + O2 + reduced [NADPH--
CC         hemoprotein reductase] = 4'-demethylepipodophyllotoxin + H(+) + H2O +
CC         oxidized [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:49032,
CC         Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:1729,
CC         ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC         ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:74422;
CC         EC=1.14.14.132; Evidence={ECO:0000269|PubMed:26359402};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49033;
CC         Evidence={ECO:0000269|PubMed:26359402};
CC   -!- COFACTOR:
CC       Name=heme; Xref=ChEBI:CHEBI:30413;
CC         Evidence={ECO:0000250|UniProtKB:Q96242};
CC   -!- PATHWAY: Aromatic compound metabolism; phenylpropanoid biosynthesis.
CC       {ECO:0000269|PubMed:26359402}.
CC   -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC       protein {ECO:0000255}.
CC   -!- TISSUE SPECIFICITY: Rhizome-specific expression.
CC       {ECO:0000269|PubMed:26359402}.
CC   -!- BIOTECHNOLOGY: Combinatorially expression of Sinopodophyllum hexandrum
CC       (mayapple) genes of the podophyllotoxin pathway (e.g. DIR, PLR, SDH,
CC       CYP719A23, OMT3, CYP71CU1, OMT1, 2-ODD, CYP71BE54 and CYP82D61) in
CC       Nicotiana benthamiana (tobacco) results in the production of the
CC       chemotherapeutic compound etoposide. {ECO:0000305|PubMed:26359402}.
CC   -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR   EMBL; KT390182; ALG05144.1; -; mRNA.
DR   AlphaFoldDB; A0A0N7F297; -.
DR   SMR; A0A0N7F297; -.
DR   KEGG; ag:ALG05144; -.
DR   BRENDA; 1.14.14.132; 4928.
DR   UniPathway; UPA00711; -.
DR   GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR   GO; GO:0020037; F:heme binding; IEA:InterPro.
DR   GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR   GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR   GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IDA:UniProtKB.
DR   GO; GO:0019438; P:aromatic compound biosynthetic process; IDA:UniProtKB.
DR   GO; GO:0009699; P:phenylpropanoid biosynthetic process; IDA:UniProtKB.
DR   Gene3D; 1.10.630.10; -; 1.
DR   InterPro; IPR001128; Cyt_P450.
DR   InterPro; IPR017972; Cyt_P450_CS.
DR   InterPro; IPR002401; Cyt_P450_E_grp-I.
DR   InterPro; IPR036396; Cyt_P450_sf.
DR   Pfam; PF00067; p450; 1.
DR   PRINTS; PR00463; EP450I.
DR   PRINTS; PR00385; P450.
DR   SUPFAM; SSF48264; SSF48264; 1.
DR   PROSITE; PS00086; CYTOCHROME_P450; 1.
PE   1: Evidence at protein level;
KW   Heme; Iron; Membrane; Metal-binding; Monooxygenase; Oxidoreductase;
KW   Transmembrane; Transmembrane helix.
FT   CHAIN           1..519
FT                   /note="Demethylepipodophyllotoxin synthase"
FT                   /id="PRO_0000451903"
FT   TRANSMEM        6..26
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   BINDING         458
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|UniProtKB:Q96242"
SQ   SEQUENCE   519 AA;  59010 MW;  1FA96EFA7CF0A0F1 CRC64;
     MDSLHCLETL LLGFFVLLPC FFYFVWKKPN NKIKEPPQPA GAWPIIGHLH LLARGDLPHK
     ILSSFADKNG PVFKIQLGVH QALVVNNSEI AKECFTTNDR FFLNRPSGVA AKIMGYNYVM
     LGVAPYGPYW RDMRKIIMLE FLSNRRLQSL KHVWHSEISI SSKELYKLWE TQNIDFCLVD
     MKQWLADLTL NMSVKMVVGK RFFGSASASA CEETESSNCP KTLRNMFRLM GSFVLSDYLP
     YLRWLDLGGH EKEMKRTVKE LDILFKGWLD EHKRKRLSGG KEDDDQDFMD VMLSILEESK
     LGNDVDTINK TACLAIILGG ADTTWATLTW ALSLLLNNPN ALKKAQDELD LHVGRDRNVD
     ESDLVKLTYI DAIIKETLRL YPPGPLLGPR VVTEDCTIAG YHVRAGTRLI VNAWKIQRDP
     LVWSQPHEYQ PERFLERDVD MKGQHFELIP FGSGRRACPA ISLALQVLPL TLAHILHGFE
     LRTPNQNKVD MTETPGIVHA KATPLEVLVA PRISPKCFV