ERYK_SACEN
ID ERYK_SACEN Reviewed; 397 AA.
AC P48635; A4F7N0; O33990;
DT 01-FEB-1996, integrated into UniProtKB/Swiss-Prot.
DT 22-FEB-2012, sequence version 3.
DT 03-AUG-2022, entry version 116.
DE RecName: Full=Erythromycin C-12 hydroxylase;
DE EC=1.14.13.154;
DE AltName: Full=Cytochrome P450 113A1;
DE Short=CYP113A1;
DE AltName: Full=Erythromycin D C-12 hydroxylase;
GN Name=eryK; Synonyms=CYP113A1; OrderedLocusNames=SACE_0713;
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], FUNCTION IN ERYTHROMYCIN BIOSYNTHESIS,
RP AND DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC 2338;
RX PubMed=8416893; DOI=10.1128/jb.175.1.182-189.1993;
RA Stassi D., Donadio S., Staver M.J., Katz L.;
RT "Identification of a Saccharopolyspora erythraea gene required for the
RT final hydroxylation step in erythromycin biosynthesis.";
RL J. Bacteriol. 175:182-189(1993).
RN [2]
RP SEQUENCE REVISION.
RC STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC 2338;
RX PubMed=9249068; DOI=10.1016/s0378-1119(97)00086-3;
RA Pereda A., Summers R., Katz L.;
RT "Nucleotide sequence of the ermE distal flank of the erythromycin
RT biosynthesis cluster in Saccharopolyspora erythraea.";
RL Gene 193:65-71(1997).
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, COFACTOR, SUBSTRATE SPECIFICITY, KINETIC
RP PARAMETERS, PATHWAY, AND SUBUNIT.
RC STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC 2338;
RX PubMed=7849045; DOI=10.1021/bi00006a006;
RA Lambalot R.H., Cane D.E., Aparicio J.J., Katz L.;
RT "Overproduction and characterization of the erythromycin C-12 hydroxylase,
RT EryK.";
RL Biochemistry 34:1858-1866(1995).
RN [5]
RP CRYSTALLIZATION.
RX PubMed=19075827; DOI=10.2174/092986608786071201;
RA Savino C., Sciara G., Miele A.E., Kendrew S.G., Vallone B.;
RT "Cloning, expression, purification, crystallization and preliminary X-ray
RT crystallographic analysis of C-12 hydroxylase EryK from Saccharopolyspora
RT erythraea.";
RL Protein Pept. Lett. 15:1138-1141(2008).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.59 ANGSTROMS) OF 2-397 IN COMPLEXES WITH HEME AND
RP ERYTHROMYCIN D, COFACTOR, AND SUBUNIT.
RX PubMed=19625248; DOI=10.1074/jbc.m109.003590;
RA Savino C., Montemiglio L.C., Sciara G., Miele A.E., Kendrew S.G., Jemth P.,
RA Gianni S., Vallone B.;
RT "Investigating the structural plasticity of a cytochrome P450: three-
RT dimensional structures of P450 EryK and binding to its physiological
RT substrate.";
RL J. Biol. Chem. 284:29170-29179(2009).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF 2-397 IN COMPLEXES WITH HEME AND
RP THE INHIBITORS KETOCONAZOLE AND CLOTRIMAZOLE, AND COFACTOR.
RX PubMed=20845962; DOI=10.1021/bi101062v;
RA Montemiglio L.C., Gianni S., Vallone B., Savino C.;
RT "Azole drugs trap cytochrome P450 EryK in alternative conformational
RT states.";
RL Biochemistry 49:9199-9206(2010).
CC -!- FUNCTION: Responsible for the C-12 hydroxylation of the macrolactone
CC ring of erythromycin. Thus, EryK catalyzes the hydroxylation of
CC erythromycin D (ErD) at the C-12 position to produce erythromycin C
CC (ErC). Erythromycin B (ErB) is not a substrate for this enzyme.
CC {ECO:0000269|PubMed:7849045, ECO:0000269|PubMed:8416893}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=erythromycin D + H(+) + NADPH + O2 = erythromycin C + H2O +
CC NADP(+); Xref=Rhea:RHEA:32631, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:63677, ChEBI:CHEBI:64258; EC=1.14.13.154;
CC Evidence={ECO:0000269|PubMed:7849045};
CC -!- COFACTOR:
CC Name=heme b; Xref=ChEBI:CHEBI:60344;
CC Evidence={ECO:0000269|PubMed:19625248, ECO:0000269|PubMed:20845962,
CC ECO:0000269|PubMed:7849045};
CC Note=Binds 1 heme b (iron(II)-protoporphyrin IX) group per subunit.
CC {ECO:0000269|PubMed:19625248, ECO:0000269|PubMed:20845962,
CC ECO:0000269|PubMed:7849045};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8.0 uM for erythromycin D {ECO:0000269|PubMed:7849045};
CC Note=kcat is 108 min(-1).;
CC -!- PATHWAY: Antibiotic biosynthesis; erythromycin biosynthesis.
CC {ECO:0000269|PubMed:7849045}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:19625248,
CC ECO:0000269|PubMed:7849045}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene no longer produce
CC erythromycin A but accumulate the B and D forms of the antibiotic.
CC {ECO:0000269|PubMed:8416893}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; U82823; AAC45584.1; -; Genomic_DNA.
DR EMBL; AM420293; CAM00054.1; -; Genomic_DNA.
DR PIR; B40634; B40634.
DR RefSeq; WP_009950895.1; NZ_PDBV01000001.1.
DR PDB; 2JJN; X-ray; 1.59 A; A=1-397.
DR PDB; 2JJO; X-ray; 1.99 A; A=1-397.
DR PDB; 2JJP; X-ray; 2.10 A; A=1-397.
DR PDB; 2WIO; X-ray; 2.00 A; A=2-397.
DR PDB; 2XFH; X-ray; 1.90 A; A=2-397.
DR PDB; 3ZKP; X-ray; 2.00 A; A=2-397.
DR PDBsum; 2JJN; -.
DR PDBsum; 2JJO; -.
DR PDBsum; 2JJP; -.
DR PDBsum; 2WIO; -.
DR PDBsum; 2XFH; -.
DR PDBsum; 3ZKP; -.
DR AlphaFoldDB; P48635; -.
DR SMR; P48635; -.
DR STRING; 405948.SACE_0713; -.
DR EnsemblBacteria; CAM00054; CAM00054; SACE_0713.
DR KEGG; sen:SACE_0713; -.
DR eggNOG; COG2124; Bacteria.
DR HOGENOM; CLU_033716_0_2_11; -.
DR OMA; VREMNGY; -.
DR OrthoDB; 816674at2; -.
DR BioCyc; MetaCyc:MON-17062; -.
DR BRENDA; 1.14.13.154; 5518.
DR BRENDA; 1.14.15.33; 5518.
DR UniPathway; UPA00240; -.
DR EvolutionaryTrace; P48635; -.
DR Proteomes; UP000006728; Chromosome.
DR GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IMP:UniProtKB.
DR GO; GO:0050661; F:NADP binding; IDA:UniProtKB.
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:0033068; P:macrolide 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; Heme; Iron; Metal-binding;
KW Monooxygenase; NADP; Oxidoreductase; Reference proteome.
FT CHAIN 1..397
FT /note="Erythromycin C-12 hydroxylase"
FT /id="PRO_0000052229"
FT REGION 307..326
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 307..321
FT /note="Basic and acidic residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 74..75
FT /ligand="substrate"
FT BINDING 81
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT BINDING 85
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT BINDING 278
FT /ligand="substrate"
FT BINDING 279
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT BINDING 337
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT BINDING 339
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 CONFLICT 330
FT /note="L -> F (in Ref. 1 and 2; AAC45584)"
FT /evidence="ECO:0000305"
FT STRAND 10..12
FT /evidence="ECO:0007829|PDB:2WIO"
FT HELIX 13..26
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 28..31
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 37..39
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 42..50
FT /evidence="ECO:0007829|PDB:2JJN"
FT TURN 52..54
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 55..57
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 59..62
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 73..75
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 80..91
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 94..98
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 101..113
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 117..120
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 121..124
FT /evidence="ECO:0007829|PDB:2JJN"
FT TURN 125..127
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 128..138
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 142..144
FT /evidence="ECO:0007829|PDB:2XFH"
FT HELIX 151..157
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 166..192
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 198..204
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 207..209
FT /evidence="ECO:0007829|PDB:2WIO"
FT HELIX 214..245
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 248..255
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 257..259
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 260..270
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 276..283
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 285..287
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 290..292
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 297..301
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 302..306
FT /evidence="ECO:0007829|PDB:2JJN"
FT TURN 309..311
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 312..314
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 326..328
FT /evidence="ECO:0007829|PDB:2XFH"
FT HELIX 330..332
FT /evidence="ECO:0007829|PDB:2JJN"
FT HELIX 342..360
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 363..365
FT /evidence="ECO:0007829|PDB:2JJO"
FT STRAND 377..383
FT /evidence="ECO:0007829|PDB:2JJN"
FT STRAND 386..388
FT /evidence="ECO:0007829|PDB:2JJN"
SQ SEQUENCE 397 AA; 43725 MW; 0106AF77AFAC86F0 CRC64;
MTTIDEVPGM ADETALLDWL GTMREKQPVW QDRYGVWHVF RHADVQTVLR DTATFSSDPT
RVIEGASPTP GMIHEIDPPE HRALRKVVSS AFTPRTISDL EPRIRDVTRS LLADAGESFD
LVDVLAFPLP VTIVAELLGL PPMDHEQFGD WSGALVDIQM DDPTDPALAE RIADVLNPLT
AYLKARCAER RADPGDDLIS RLVLAEVDGR ALDDEEAANF STALLLAGHI TTTVLLGNIV
RTLDEHPAHW DAAAEDPGRI PAIVEEVLRY RPPFPQMQRT TTKATEVAGV PIPADVMVNT
WVLSANRDSD AHDDPDRFDP SRKSGGAAQL SFGHGVHFCL GAPLARLENR VALEEIIARF
GRLTVDRDDE RLRHFEQIVL GTRHLPVLAG SSPRQSA
