CPXJ_SACEN
ID CPXJ_SACEN Reviewed; 404 AA.
AC Q00441; A4F7P7;
DT 01-FEB-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1995, sequence version 2.
DT 03-AUG-2022, entry version 146.
DE RecName: Full=6-deoxyerythronolide B hydroxylase {ECO:0000303|PubMed:2011746};
DE Short=6-DEB hydroxylase {ECO:0000303|PubMed:2011746};
DE EC=1.14.15.35 {ECO:0000269|PubMed:1732208, ECO:0000269|PubMed:2011746};
DE AltName: Full=CYPCVIIA1;
DE AltName: Full=Cytochrome P450 107A1;
DE AltName: Full=Cytochrome P450eryF {ECO:0000303|PubMed:10716705, ECO:0000303|PubMed:15824115, ECO:0000303|PubMed:7749919};
DE AltName: Full=Erythromycin A biosynthesis hydroxylase;
GN Name=eryF {ECO:0000303|PubMed:2011746}; Synonyms=CYP107A1;
GN OrderedLocusNames=SACE_0730;
OS Saccharopolyspora erythraea (strain ATCC 11635 / DSM 40517 / JCM 4748 /
OS NBRC 13426 / NCIMB 8594 / NRRL 2338).
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC Saccharopolyspora.
OX NCBI_TaxID=405948;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=1840640; DOI=10.1007/bf00290659;
RA Haydock S.F., Dowson J.A., Dhillon N., Roberts G.A., Cortes J.,
RA Leadlay P.F.;
RT "Cloning and sequence analysis of genes involved in erythromycin
RT biosynthesis in Saccharopolyspora erythraea: sequence similarities between
RT EryG and a family of S-adenosylmethionine-dependent methyltransferases.";
RL Mol. Gen. Genet. 230:120-128(1991).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, DISRUPTION
RP PHENOTYPE, AND PATHWAY.
RX PubMed=2011746; DOI=10.1126/science.2011746;
RA Weber J.M., Leung J.O., Swanson S.J., Idler K.B., McAlpine J.B.;
RT "An erythromycin derivative produced by targeted gene disruption in
RT Saccharopolyspora erythraea.";
RL Science 252:114-117(1991).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC 2338;
RX PubMed=17369815; DOI=10.1038/nbt1297;
RA Oliynyk M., Samborskyy M., Lester J.B., Mironenko T., Scott N., Dickens S.,
RA Haydock S.F., Leadlay P.F.;
RT "Complete genome sequence of the erythromycin-producing bacterium
RT Saccharopolyspora erythraea NRRL23338.";
RL Nat. Biotechnol. 25:447-453(2007).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=1732208; DOI=10.1128/jb.174.3.725-735.1992;
RA Andersen J.F., Hutchinson C.R.;
RT "Characterization of Saccharopolyspora erythraea cytochrome P-450 genes and
RT enzymes, including 6-deoxyerythronolide B hydroxylase.";
RL J. Bacteriol. 174:725-735(1992).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 3-404 IN COMPLEX WITH HEME AND
RP 6-DEOXYERYTHRONOLIDE B, AND COFACTOR.
RX PubMed=7749919; DOI=10.1038/nsb0295-144;
RA Cupp-Vickery J.L., Poulos T.L.;
RT "Structure of cytochrome P450eryF involved in erythromycin biosynthesis.";
RL Nat. Struct. Biol. 2:144-153(1995).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 3-404 IN COMPLEX WITH HEME, AND
RP COFACTOR.
RX PubMed=10716705; DOI=10.1073/pnas.97.7.3050;
RA Cupp-Vickery J., Anderson R., Hatziris Z.;
RT "Crystal structures of ligand complexes of P450eryF exhibiting homotropic
RT cooperativity.";
RL Proc. Natl. Acad. Sci. U.S.A. 97:3050-3055(2000).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 2-404 IN COMPLEX WITH HEME, AND
RP COFACTOR.
RX PubMed=11469860; DOI=10.1006/jmbi.2001.4803;
RA Cupp-Vickery J., Garcia C., Hofacre A., McGee-Estrada K.;
RT "Ketoconazole-induced conformational changes in the active site of
RT cytochrome P450eryF.";
RL J. Mol. Biol. 311:101-110(2001).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH HEME AND
RP 6-DEOXYERYTHRONOLIDE B, AND COFACTOR.
RX PubMed=15824115; DOI=10.1074/jbc.m501732200;
RA Nagano S., Cupp-Vickery J.R., Poulos T.L.;
RT "Crystal structures of the ferrous dioxygen complex of wild-type cytochrome
RT P450eryF and its mutants, A245S and A245T: investigation of the proton
RT transfer system in P450eryF.";
RL J. Biol. Chem. 280:22102-22107(2005).
CC -!- FUNCTION: Catalyzes the conversion of 6-deoxyerythronolide B (6-DEB) to
CC erythronolide B (EB) by the insertion of an oxygen at the 6S position
CC of 6-DEB. Requires the participation of a ferredoxin and a ferredoxin
CC reductase for the transfer of electrons from NADPH to the
CC monooxygenase. {ECO:0000269|PubMed:1732208,
CC ECO:0000269|PubMed:2011746}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=6-deoxyerythronolide B + 2 H(+) + O2 + 2 reduced [2Fe-2S]-
CC [ferredoxin] = erythronolide B + H2O + 2 oxidized [2Fe-2S]-
CC [ferredoxin]; Xref=Rhea:RHEA:40299, Rhea:RHEA-COMP:10000, Rhea:RHEA-
CC COMP:10001, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16089, ChEBI:CHEBI:27977, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738; EC=1.14.15.35;
CC Evidence={ECO:0000269|PubMed:1732208, ECO:0000269|PubMed:2011746};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000269|PubMed:10716705, ECO:0000269|PubMed:11469860,
CC ECO:0000269|PubMed:7749919};
CC -!- PATHWAY: Antibiotic biosynthesis; erythromycin biosynthesis.
CC {ECO:0000269|PubMed:1732208, ECO:0000269|PubMed:2011746}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:1732208}.
CC -!- DISRUPTION PHENOTYPE: Gene disruption leads to the accumulation of 6-
CC deoxyerythromycin A, an antibiotic with increased stability at low pH.
CC {ECO:0000269|PubMed:2011746}.
CC -!- MISCELLANEOUS: The bound substrate forms no direct hydrogen bonds with
CC the protein. {ECO:0000269|PubMed:15824115, ECO:0000269|PubMed:7749919}.
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; X60379; CAA42927.1; -; Genomic_DNA.
DR EMBL; M54983; AAA26496.1; -; Genomic_DNA.
DR EMBL; AM420293; CAM00071.1; -; Genomic_DNA.
DR PIR; S18531; S18531.
DR RefSeq; WP_009950397.1; NZ_PDBV01000001.1.
DR PDB; 1EGY; X-ray; 2.35 A; A=3-404.
DR PDB; 1EUP; X-ray; 2.10 A; A=3-404.
DR PDB; 1JIN; X-ray; 2.30 A; A=3-404.
DR PDB; 1JIO; X-ray; 2.10 A; A=2-404.
DR PDB; 1JIP; X-ray; 2.00 A; A=2-404.
DR PDB; 1OXA; X-ray; 2.10 A; A=3-404.
DR PDB; 1Z8O; X-ray; 1.70 A; A=1-404.
DR PDB; 1Z8P; X-ray; 1.85 A; A=1-404.
DR PDB; 1Z8Q; X-ray; 2.00 A; A=1-404.
DR PDBsum; 1EGY; -.
DR PDBsum; 1EUP; -.
DR PDBsum; 1JIN; -.
DR PDBsum; 1JIO; -.
DR PDBsum; 1JIP; -.
DR PDBsum; 1OXA; -.
DR PDBsum; 1Z8O; -.
DR PDBsum; 1Z8P; -.
DR PDBsum; 1Z8Q; -.
DR AlphaFoldDB; Q00441; -.
DR SMR; Q00441; -.
DR STRING; 405948.SACE_0730; -.
DR DrugBank; DB04070; 6-Deoxyerythronolide B.
DR DrugBank; DB03369; 9-Aminophenanthrene.
DR DrugBank; DB01536; Androstenedione.
DR PRIDE; Q00441; -.
DR EnsemblBacteria; CAM00071; CAM00071; SACE_0730.
DR KEGG; sen:SACE_0730; -.
DR eggNOG; COG2124; Bacteria.
DR HOGENOM; CLU_033716_1_0_11; -.
DR OMA; QLKIIWE; -.
DR OrthoDB; 816674at2; -.
DR BRENDA; 1.14.15.35; 5518.
DR UniPathway; UPA00240; -.
DR EvolutionaryTrace; Q00441; -.
DR Proteomes; UP000006728; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IDA:UniProtKB.
DR GO; GO:0004497; F:monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR GO; GO:1901115; P:erythromycin 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 SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Cytoplasm; Heme; Iron;
KW Metal-binding; Monooxygenase; Oxidoreductase; Reference proteome.
FT CHAIN 1..404
FT /note="6-deoxyerythronolide B hydroxylase"
FT /id="PRO_0000052220"
FT BINDING 351
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:0000269|PubMed:10716705,
FT ECO:0000269|PubMed:11469860, ECO:0000269|PubMed:15824115,
FT ECO:0000269|PubMed:7749919"
FT CONFLICT 261
FT /note="P -> PDQ (in Ref. 1; CAA42927)"
FT /evidence="ECO:0000305"
FT CONFLICT 302
FT /note="G -> R (in Ref. 1; CAA42927)"
FT /evidence="ECO:0000305"
FT HELIX 10..12
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 16..26
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 28..34
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 37..42
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 45..53
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 73..75
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 80..86
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 90..92
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 97..106
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 111..116
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 118..130
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 134..139
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 140..143
FT /evidence="ECO:0007829|PDB:1Z8O"
FT TURN 144..146
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 147..156
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 161..163
FT /evidence="ECO:0007829|PDB:1Z8O"
FT TURN 164..166
FT /evidence="ECO:0007829|PDB:1JIO"
FT HELIX 167..175
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 179..181
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 182..205
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 211..217
FT /evidence="ECO:0007829|PDB:1Z8O"
FT TURN 221..223
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 228..259
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 261..269
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 271..273
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 274..284
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 291..297
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 299..301
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 304..306
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 311..314
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 316..319
FT /evidence="ECO:0007829|PDB:1Z8O"
FT TURN 323..325
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 326..328
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 334..336
FT /evidence="ECO:0007829|PDB:1EGY"
FT STRAND 342..344
FT /evidence="ECO:0007829|PDB:1EGY"
FT HELIX 347..349
FT /evidence="ECO:0007829|PDB:1EUP"
FT HELIX 354..371
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 376..379
FT /evidence="ECO:0007829|PDB:1Z8O"
FT HELIX 381..383
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 390..392
FT /evidence="ECO:0007829|PDB:1Z8O"
FT STRAND 399..401
FT /evidence="ECO:0007829|PDB:1Z8O"
SQ SEQUENCE 404 AA; 45099 MW; 257ABEFC2D88B3FE CRC64;
MTTVPDLESD SFHVDWYRTY AELRETAPVT PVRFLGQDAW LVTGYDEAKA ALSDLRLSSD
PKKKYPGVEV EFPAYLGFPE DVRNYFATNM GTSDPPTHTR LRKLVSQEFT VRRVEAMRPR
VEQITAELLD EVGDSGVVDI VDRFAHPLPI KVICELLGVD EKYRGEFGRW SSEILVMDPE
RAEQRGQAAR EVVNFILDLV ERRRTEPGDD LLSALIRVQD DDDGRLSADE LTSIALVLLL
AGFEASVSLI GIGTYLLLTH PDQLALVRRD PSALPNAVEE ILRYIAPPET TTRFAAEEVE
IGGVAIPQYS TVLVANGAAN RDPKQFPDPH RFDVTRDTRG HLSFGQGIHF CMGRPLAKLE
GEVALRALFG RFPALSLGID ADDVVWRRSL LLRGIDHLPV RLDG
