ID   CYPD_BACSU              Reviewed;        1061 AA.
AC   O08394;
DT   01-DEC-2000, integrated into UniProtKB/Swiss-Prot.
DT   01-JUL-1997, sequence version 1.
DT   27-MAR-2024, entry version 165.
DE   RecName: Full=Bifunctional cytochrome P450/NADPH--P450 reductase 1 {ECO:0000305};
DE   AltName: Full=CYP102A2 {ECO:0000303|PubMed:15122913};
DE   AltName: Full=Fatty acid hydroxylase CypD {ECO:0000305};
DE   AltName: Full=Flavocytochrome P450 102A2 {ECO:0000305};
DE   Includes:
DE     RecName: Full=Cytochrome P450 102A2 {ECO:0000305};
DE              EC=1.14.14.1 {ECO:0000269|PubMed:15122913, ECO:0000269|PubMed:15375636};
DE   Includes:
DE     RecName: Full=NADPH--cytochrome P450 reductase {ECO:0000305};
DE              EC=1.6.2.4 {ECO:0000269|PubMed:15122913, ECO:0000269|PubMed:15375636};
GN   Name=cypD {ECO:0000312|EMBL:CAB12544.1};
GN   Synonyms=cyp102A2 {ECO:0000303|PubMed:15122913,
GN   ECO:0000303|PubMed:15375636}, yetO {ECO:0000303|PubMed:15122913},
GN   yfnJ; OrderedLocusNames=BSU07250;
OS   Bacillus subtilis (strain 168).
OC   Bacteria; Bacillota; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX   NCBI_TaxID=224308;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9308178; DOI=10.1099/00221287-143-9-2939;
RA   Sorokin A., Bolotin A., Purnelle B., Hilbert H., Lauber J.,
RA   Duesterhoeft A., Ehrlich S.D.;
RT   "Sequence of the Bacillus subtilis genome region in the vicinity of the lev
RT   operon reveals two new extracytoplasmic function RNA polymerase sigma
RT   factors SigV and SigZ.";
RL   Microbiology 143:2939-2943(1997).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9384377; DOI=10.1038/36786;
RA   Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA   Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA   Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA   Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA   Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA   Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA   Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA   Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA   Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA   Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA   Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA   Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA   Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA   Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA   Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA   Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA   Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA   Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA   Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA   Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA   Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA   Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA   Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA   Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA   Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA   Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA   Yoshikawa H., Danchin A.;
RT   "The complete genome sequence of the Gram-positive bacterium Bacillus
RT   subtilis.";
RL   Nature 390:249-256(1997).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND BIOPHYSICOCHEMICAL
RP   PROPERTIES.
RX   PubMed=15375636; DOI=10.1007/s00253-004-1719-y;
RA   Budde M., Maurer S.C., Schmid R.D., Urlacher V.B.;
RT   "Cloning, expression and characterisation of CYP102A2, a self-sufficient
RT   P450 monooxygenase from Bacillus subtilis.";
RL   Appl. Microbiol. Biotechnol. 66:180-186(2004).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE SPECIFICITY, AND
RP   BIOPHYSICOCHEMICAL PROPERTIES.
RC   STRAIN=168 / 1A1;
RX   PubMed=15122913; DOI=10.1021/bi035904m;
RA   Gustafsson M.C., Roitel O., Marshall K.R., Noble M.A., Chapman S.K.,
RA   Pessegueiro A., Fulco A.J., Cheesman M.R., von Wachenfeldt C., Munro A.W.;
RT   "Expression, purification, and characterization of Bacillus subtilis
RT   cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of P450
RT   BM3 from Bacillus megaterium.";
RL   Biochemistry 43:5474-5487(2004).
RN   [5]
RP   PROTEIN ENGINEERING, AND MUTAGENESIS OF PRO-15.
RC   STRAIN=168;
RX   PubMed=15857787; DOI=10.1016/j.bioeng.2004.11.003;
RA   Axarli I., Prigipaki A., Labrou N.E.;
RT   "Engineering the substrate specificity of cytochrome P450 CYP102A2 by
RT   directed evolution: production of an efficient enzyme for bioconversion of
RT   fine chemicals.";
RL   Biomol. Eng. 22:81-88(2005).
RN   [6]
RP   FUNCTION.
RX   PubMed=21048857; DOI=10.4061/2010/125429;
RA   Axarli I., Prigipaki A., Labrou N.E.;
RT   "Cytochrome P450 102A2 catalyzes efficient oxidation of sodium dodecyl
RT   sulphate: a molecular tool for remediation.";
RL   Enzyme Res. 2010:125429-125429(2010).
CC   -!- FUNCTION: Functions as a fatty acid monooxygenase. Catalyzes
CC       hydroxylation of a range of long-chain fatty acids, with a preference
CC       for long-chain unsaturated and branched-chain fatty acids over
CC       saturated fatty acids. Hydroxylation of myristic acid occurs mainly at
CC       the omega-2 position. Also displays a NADPH-dependent reductase
CC       activity in the C-terminal domain, which allows electron transfer from
CC       NADPH to the heme iron of the cytochrome P450 N-terminal domain
CC       (PubMed:15375636, PubMed:15122913). Is also able to catalyze efficient
CC       oxidation of sodium dodecyl sulfate (SDS) (PubMed:21048857).
CC       {ECO:0000269|PubMed:15122913, ECO:0000269|PubMed:15375636,
CC       ECO:0000269|PubMed:21048857}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=an organic molecule + O2 + reduced [NADPH--hemoprotein
CC         reductase] = an alcohol + H(+) + H2O + oxidized [NADPH--hemoprotein
CC         reductase]; Xref=Rhea:RHEA:17149, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC         COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC         ChEBI:CHEBI:30879, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC         ChEBI:CHEBI:142491; EC=1.14.14.1;
CC         Evidence={ECO:0000269|PubMed:15122913, ECO:0000269|PubMed:15375636};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=NADPH + 2 oxidized [cytochrome P450] = H(+) + NADP(+) + 2
CC         reduced [cytochrome P450]; Xref=Rhea:RHEA:24040, Rhea:RHEA-
CC         COMP:14627, Rhea:RHEA-COMP:14628, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:55376, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC         ChEBI:CHEBI:60344; EC=1.6.2.4; Evidence={ECO:0000269|PubMed:15122913,
CC         ECO:0000269|PubMed:15375636};
CC   -!- COFACTOR:
CC       Name=FAD; Xref=ChEBI:CHEBI:57692;
CC         Evidence={ECO:0000269|PubMed:15122913};
CC   -!- COFACTOR:
CC       Name=FMN; Xref=ChEBI:CHEBI:58210;
CC         Evidence={ECO:0000269|PubMed:15122913};
CC   -!- COFACTOR:
CC       Name=heme b; Xref=ChEBI:CHEBI:60344;
CC         Evidence={ECO:0000269|PubMed:15122913};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=38.8 uM for stearic acid {ECO:0000269|PubMed:15122913};
CC         KM=150 uM for phytanic acid {ECO:0000269|PubMed:15122913};
CC         KM=56.8 uM for 15-methylpalmitic acid {ECO:0000269|PubMed:15122913};
CC         KM=3.6 uM for NADPH {ECO:0000269|PubMed:15122913};
CC         KM=17.9 mM for NADH {ECO:0000269|PubMed:15122913};
CC         KM=6.9 uM for cytochrome c (in the reductase assay)
CC         {ECO:0000269|PubMed:15122913};
CC         KM=153.4 uM for ferricyanide (in the reductase assay)
CC         {ECO:0000269|PubMed:15122913};
CC         KM=17.36 uM for oleic acid {ECO:0000269|PubMed:15375636};
CC         Note=kcat is 430 min(-1) for stearic acid hydroxylation. kcat is 5430
CC         min(-1) for phytanic acid hydroxylation. kcat is 6105 min(-1) for
CC         15-methylpalmitic acid hydroxylation. kcat is 11400 min(-1) for the
CC         reduction of cytochrome c. kcat is 38150 min(-1) for the reduction of
CC         ferricyanide (PubMed:15122913). kcat is 2244 min(-1) for oleic acid
CC         hydroxylation (PubMed:15375636). {ECO:0000269|PubMed:15122913,
CC         ECO:0000269|PubMed:15375636};
CC       pH dependence:
CC         Optimum pH is 7.0. {ECO:0000269|PubMed:15375636};
CC       Temperature dependence:
CC         Optimum temperature is 51 degrees Celsius. However, enzyme stability
CC         is dramatically reduced at this temperature, incubation for 30
CC         minutes at 31 and 49 degrees Celsius results in 61% and 17% residual
CC         activity, respectively. Incubation at 60 degrees Celsius leads to
CC         total inactivation of the enzyme. {ECO:0000269|PubMed:15375636};
CC   -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC   -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome P450
CC       family. {ECO:0000305}.
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DR   EMBL; D87979; BAA20123.1; -; Genomic_DNA.
DR   EMBL; AL009126; CAB12544.1; -; Genomic_DNA.
DR   PIR; D69799; D69799.
DR   RefSeq; NP_388606.1; NC_000964.3.
DR   RefSeq; WP_003242884.1; NZ_JNCM01000032.1.
DR   AlphaFoldDB; O08394; -.
DR   SMR; O08394; -.
DR   STRING; 224308.BSU07250; -.
DR   PaxDb; 224308-BSU07250; -.
DR   EnsemblBacteria; CAB12544; CAB12544; BSU_07250.
DR   GeneID; 938784; -.
DR   KEGG; bsu:BSU07250; -.
DR   PATRIC; fig|224308.179.peg.786; -.
DR   eggNOG; COG0369; Bacteria.
DR   eggNOG; COG2124; Bacteria.
DR   InParanoid; O08394; -.
DR   OrthoDB; 9789468at2; -.
DR   PhylomeDB; O08394; -.
DR   BioCyc; BSUB:BSU07250-MONOMER; -.
DR   SABIO-RK; O08394; -.
DR   Proteomes; UP000001570; Chromosome.
DR   GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR   GO; GO:0070330; F:aromatase activity; IEA:UniProtKB-EC.
DR   GO; GO:0005504; F:fatty acid binding; IDA:UniProtKB.
DR   GO; GO:0050660; F:flavin adenine dinucleotide binding; IDA:UniProtKB.
DR   GO; GO:0010181; F:FMN binding; IDA:UniProtKB.
DR   GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR   GO; GO:0005506; F:iron ion binding; ISS:UniProtKB.
DR   GO; GO:0003958; F:NADPH-hemoprotein reductase activity; IDA:UniProtKB.
DR   GO; GO:0016491; F:oxidoreductase activity; IBA:GO_Central.
DR   GO; GO:0016712; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR   GO; GO:0019395; P:fatty acid oxidation; IDA:UniProtKB.
DR   CDD; cd06206; bifunctional_CYPOR; 1.
DR   CDD; cd11068; CYP120A1; 1.
DR   Gene3D; 3.40.50.360; -; 1.
DR   Gene3D; 1.10.630.10; Cytochrome P450; 1.
DR   Gene3D; 3.40.50.80; Nucleotide-binding domain of ferredoxin-NADP reductase (FNR) module; 1.
DR   Gene3D; 2.40.30.10; Translation factors; 1.
DR   InterPro; IPR023206; Bifunctional_P450_P450_red.
DR   InterPro; IPR003097; CysJ-like_FAD-binding.
DR   InterPro; IPR001128; Cyt_P450.
DR   InterPro; IPR017972; Cyt_P450_CS.
DR   InterPro; IPR036396; Cyt_P450_sf.
DR   InterPro; IPR017927; FAD-bd_FR_type.
DR   InterPro; IPR001094; Flavdoxin-like.
DR   InterPro; IPR008254; Flavodoxin/NO_synth.
DR   InterPro; IPR001709; Flavoprot_Pyr_Nucl_cyt_Rdtase.
DR   InterPro; IPR029039; Flavoprotein-like_sf.
DR   InterPro; IPR039261; FNR_nucleotide-bd.
DR   InterPro; IPR023173; NADPH_Cyt_P450_Rdtase_alpha.
DR   InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR   InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR   PANTHER; PTHR19384:SF17; NADPH--CYTOCHROME P450 REDUCTASE; 1.
DR   PANTHER; PTHR19384; NITRIC OXIDE SYNTHASE-RELATED; 1.
DR   Pfam; PF00667; FAD_binding_1; 1.
DR   Pfam; PF00258; Flavodoxin_1; 1.
DR   Pfam; PF00175; NAD_binding_1; 1.
DR   Pfam; PF00067; p450; 1.
DR   PIRSF; PIRSF000209; Bifunctional_P450_P450R; 1.
DR   PRINTS; PR00369; FLAVODOXIN.
DR   PRINTS; PR00371; FPNCR.
DR   SUPFAM; SSF48264; Cytochrome P450; 1.
DR   SUPFAM; SSF52343; Ferredoxin reductase-like, C-terminal NADP-linked domain; 1.
DR   SUPFAM; SSF52218; Flavoproteins; 1.
DR   SUPFAM; SSF63380; Riboflavin synthase domain-like; 1.
DR   PROSITE; PS00086; CYTOCHROME_P450; 1.
DR   PROSITE; PS51384; FAD_FR; 1.
DR   PROSITE; PS50902; FLAVODOXIN_LIKE; 1.
PE   1: Evidence at protein level;
KW   Cytoplasm; Electron transport; FAD; Flavoprotein; FMN; Heme; Iron;
KW   Metal-binding; Monooxygenase; Multifunctional enzyme; NADP; Oxidoreductase;
KW   Reference proteome; Transport.
FT   CHAIN           1..1061
FT                   /note="Bifunctional cytochrome P450/NADPH--P450 reductase
FT                   1"
FT                   /id="PRO_0000052206"
FT   DOMAIN          493..632
FT                   /note="Flavodoxin-like"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT   DOMAIN          671..904
FT                   /note="FAD-binding FR-type"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT   REGION          1..475
FT                   /note="Cytochrome P450"
FT                   /evidence="ECO:0000305|PubMed:15122913"
FT   REGION          476..1061
FT                   /note="NADPH--P450 reductase"
FT                   /evidence="ECO:0000305|PubMed:15122913"
FT   BINDING         403
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:P14779"
FT   BINDING         499..504
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   BINDING         546..549
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   BINDING         580..582
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   BINDING         588..590
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   SITE            271
FT                   /note="Important for catalytic activity"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   MUTAGEN         15
FT                   /note="P->S: Exhibits modified substrate specificity. Shows
FT                   approximately 6- to 9-fold increased activity with SDS,
FT                   lauric acid and 1,4-naphthoquinone, and enhanced activity
FT                   for other substrates such as ethacrynic acid and
FT                   epsilon-amino-n-caproic acid."
FT                   /evidence="ECO:0000269|PubMed:15857787"
SQ   SEQUENCE   1061 AA;  119468 MW;  7915DACC20578978 CRC64;
     MKETSPIPQP KTFGPLGNLP LIDKDKPTLS LIKLAEEQGP IFQIHTPAGT TIVVSGHELV
     KEVCDEERFD KSIEGALEKV RAFSGDGLFT SWTHEPNWRK AHNILMPTFS QRAMKDYHEK
     MVDIAVQLIQ KWARLNPNEA VDVPGDMTRL TLDTIGLCGF NYRFNSYYRE TPHPFINSMV
     RALDEAMHQM QRLDVQDKLM VRTKRQFRYD IQTMFSLVDS IIAERRANGD QDEKDLLARM
     LNVEDPETGE KLDDENIRFQ IITFLIAGHE TTSGLLSFAT YFLLKHPDKL KKAYEEVDRV
     LTDAAPTYKQ VLELTYIRMI LNESLRLWPT APAFSLYPKE DTVIGGKFPI TTNDRISVLI
     PQLHRDRDAW GKDAEEFRPE RFEHQDQVPH HAYKPFGNGQ RACIGMQFAL HEATLVLGMI
     LKYFTLIDHE NYELDIKQTL TLKPGDFHIS VQSRHQEAIH ADVQAAEKAA PDEQKEKTEA
     KGASVIGLNN RPLLVLYGSD TGTAEGVARE LADTASLHGV RTKTAPLNDR IGKLPKEGAV
     VIVTSSYNGK PPSNAGQFVQ WLQEIKPGEL EGVHYAVFGC GDHNWASTYQ YVPRFIDEQL
     AEKGATRFSA RGEGDVSGDF EGQLDEWKKS MWADAIKAFG LELNENADKE RSTLSLQFVR
     GLGESPLARS YEASHASIAE NRELQSADSD RSTRHIEIAL PPDVEYQEGD HLGVLPKNSQ
     TNVSRILHRF GLKGTDQVTL SASGRSAGHL PLGRPVSLHD LLSYSVEVQE AATRAQIREL
     ASFTVCPPHR RELEELSAEG VYQEQILKKR ISMLDLLEKY EACDMPFERF LELLRPLKPR
     YYSISSSPRV NPRQASITVG VVRGPAWSGR GEYRGVASND LAERQAGDDV VMFIRTPESR
     FQLPKDPETP IIMVGPGTGV APFRGFLQAR DVLKREGKTL GEAHLYFGCR NDRDFIYRDE
     LERFEKDGIV TVHTAFSRKE GMPKTYVQHL MADQADTLIS ILDRGGRLYV CGDGSKMAPD
     VEAALQKAYQ AVHGTGEQEA QNWLRHLQDT GMYAKDVWAG I
//