CYPD_BACSU
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 03-AUG-2022, entry version 158.
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}, yfnJ;
GN OrderedLocusNames=BSU07250;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; 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 15-
CC 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; O08394; -.
DR PRIDE; O08394; -.
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 OMA; WKKAHNI; -.
DR PhylomeDB; O08394; -.
DR BioCyc; BSUB:BSU07250-MON; -.
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 Gene3D; 1.10.630.10; -; 1.
DR Gene3D; 1.20.990.10; -; 1.
DR Gene3D; 3.40.50.360; -; 1.
DR Gene3D; 3.40.50.80; -; 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 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; SSF48264; 1.
DR SUPFAM; SSF52218; SSF52218; 1.
DR SUPFAM; SSF52343; SSF52343; 1.
DR SUPFAM; SSF63380; SSF63380; 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 epsilon-
FT 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
