ID CYPB_BACSU Reviewed; 1054 AA. AC O08336; DT 01-DEC-2000, integrated into UniProtKB/Swiss-Prot. DT 01-JUL-1997, sequence version 1. DT 27-MAR-2024, entry version 169. DE RecName: Full=Bifunctional cytochrome P450/NADPH--P450 reductase 2 {ECO:0000305}; DE AltName: Full=CYP102A3 {ECO:0000303|PubMed:15122913}; DE AltName: Full=Fatty acid hydroxylase CypB {ECO:0000305}; DE AltName: Full=Flavocytochrome P450 102A3 {ECO:0000305}; DE Includes: DE RecName: Full=Cytochrome P450 102A3 {ECO:0000305}; DE EC=1.14.14.1 {ECO:0000269|PubMed:14741768, ECO:0000269|PubMed:15122913}; DE Includes: DE RecName: Full=NADPH--cytochrome P450 reductase; DE EC=1.6.2.4 {ECO:0000269|PubMed:14741768, ECO:0000269|PubMed:15122913}; GN Name=cypB {ECO:0000312|EMBL:CAB14658.1}; GN Synonyms=cyp102A3 {ECO:0000303|PubMed:11574077, GN ECO:0000303|PubMed:15122913}, yrhJ {ECO:0000303|PubMed:11574077, GN ECO:0000303|PubMed:15122913}; OrderedLocusNames=BSU27160; 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 INDUCTION. RX PubMed=11574077; DOI=10.1093/oxfordjournals.jbchem.a003020; RA Lee T.-R., Hsu H.-P., Shaw G.-C.; RT "Transcriptional regulation of the Bacillus subtilis bscR-CYP102A3 operon RT by the BscR repressor and differential induction of cytochrome CYP102A3 RT expression by oleic acid and palmitate."; RL J. Biochem. 130:569-574(2001). RN [4] RP INDUCTION. RC STRAIN=168 / 1A1; RX PubMed=11734890; DOI=10.1007/s002030100350; RA Gustafsson M.C., Palmer C.N., Wolf C.R., von Wachenfeldt C.; RT "Fatty-acid-displaced transcriptional repressor, a conserved regulator of RT cytochrome P450 102 transcription in Bacillus species."; RL Arch. Microbiol. 176:459-464(2001). RN [5] 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 [6] RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND MUTAGENESIS OF RP PHE-89 AND SER-190. RC STRAIN=168 / ATCC 33234 / DSM 402 / NBRC 111470 / NCIMB 10106; RX PubMed=14741768; DOI=10.1016/j.jbiotec.2003.11.001; RA Lentz O., Urlacher V., Schmid R.D.; RT "Substrate specificity of native and mutated cytochrome P450 (CYP102A3) RT from Bacillus subtilis."; RL J. Biotechnol. 108:41-49(2004). RN [7] RP PROTEIN ENGINEERING. RC STRAIN=168 / ATCC 33234 / DSM 402 / NBRC 111470 / NCIMB 10106; RX PubMed=16381045; DOI=10.1002/cbic.200500266; RA Lentz O., Feenstra A., Habicher T., Hauer B., Schmid R.D., Urlacher V.B.; RT "Altering the regioselectivity of cytochrome P450 CYP102A3 of Bacillus RT subtilis by using a new versatile assay system."; RL ChemBioChem 7:345-350(2006). RN [8] RP INDUCTION. RC STRAIN=168 / 1604; RX PubMed=17434969; DOI=10.1128/jb.00130-07; RA Jervis A.J., Thackray P.D., Houston C.W., Horsburgh M.J., Moir A.; RT "SigM-responsive genes of Bacillus subtilis and their promoters."; RL J. Bacteriol. 189:4534-4538(2007). CC -!- FUNCTION: Functions as a fatty acid monooxygenase. Catalyzes CC hydroxylation of a range of medium to long-chain fatty acids, with a CC preference for long-chain unsaturated and branched-chain fatty acids CC over saturated fatty acids. Hydroxylation of myristic acid occurs CC mainly at the omega-2 and omega-3 positions, in approximately equal CC proportions. Also displays a NADPH-dependent reductase activity in the CC C-terminal domain, which allows electron transfer from NADPH to the CC heme iron of the cytochrome P450 N-terminal domain. CC {ECO:0000269|PubMed:14741768, ECO:0000269|PubMed:15122913}. 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:14741768, ECO:0000269|PubMed:15122913}; 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:14741768, CC ECO:0000269|PubMed:15122913}; 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=165 uM for lauric acid {ECO:0000269|PubMed:15122913}; CC KM=542 uM for myristic acid {ECO:0000269|PubMed:15122913}; CC KM=337 uM for palmitic acid {ECO:0000269|PubMed:15122913}; CC KM=68.5 uM for stearic acid {ECO:0000269|PubMed:15122913}; CC KM=28.7 uM for phytanic acid {ECO:0000269|PubMed:15122913}; CC KM=68.3 uM for 15-methylpalmitic acid {ECO:0000269|PubMed:15122913}; CC KM=79 uM for arachidonic acid {ECO:0000269|PubMed:15122913}; CC KM=5.1 uM for NADPH {ECO:0000269|PubMed:15122913}; CC KM=2.43 mM for NADH {ECO:0000269|PubMed:15122913}; CC KM=10.9 uM for cytochrome c (in the reductase assay) CC {ECO:0000269|PubMed:15122913}; CC KM=285 uM for ferricyanide (in the reductase assay) CC {ECO:0000269|PubMed:15122913}; CC Note=kcat is 104 min(-1) for lauric acid hydroxylation. kcat is 5556 CC min(-1) for myristic acid hydroxylation. kcat is 676 min(-1) for CC palmitic acid hydroxylation. kcat is 374 min(-1) for stearic acid CC hydroxylation. kcat is 794 min(-1) for phytanic acid hydroxylation. CC kcat is 3845 min(-1) for 15-methylpalmitic acid hydroxylation. kcat CC is 1690 min(-1) for arachidonic acid hydroxylation. kcat is 3520 CC min(-1) for the reduction of cytochrome c. kcat is 37050 min(-1) for CC the reduction of ferricyanide. {ECO:0000269|PubMed:15122913}; CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}. CC -!- INDUCTION: Negatively regulated by the transcriptional repressor FatR CC (PubMed:11574077, PubMed:11734890). Is induced by fatty acids such as CC oleate, linoleate and phytanate, that bind and displace the FatR CC repressor (PubMed:11734890). Is also induced by palmitate, likely via CC another mechanism (PubMed:11574077). Transcribed under partial control CC of SigM ECF sigma factor (PubMed:17434969). CC {ECO:0000269|PubMed:11574077, ECO:0000269|PubMed:11734890, CC ECO:0000269|PubMed:17434969}. CC -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome P450 CC 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; U93874; AAB80867.1; -; Genomic_DNA. DR EMBL; AL009126; CAB14658.1; -; Genomic_DNA. DR PIR; A69975; A69975. DR RefSeq; NP_390594.1; NC_000964.3. DR RefSeq; WP_003246174.1; NZ_JNCM01000036.1. DR AlphaFoldDB; O08336; -. DR SMR; O08336; -. DR STRING; 224308.BSU27160; -. DR PaxDb; 224308-BSU27160; -. DR DNASU; 937585; -. DR EnsemblBacteria; CAB14658; CAB14658; BSU_27160. DR GeneID; 937585; -. DR KEGG; bsu:BSU27160; -. DR PATRIC; fig|224308.179.peg.2949; -. DR eggNOG; COG0369; Bacteria. DR eggNOG; COG2124; Bacteria. DR InParanoid; O08336; -. DR OrthoDB; 9789468at2; -. DR PhylomeDB; O08336; -. DR BioCyc; BSUB:BSU27160-MONOMER; -. DR SABIO-RK; O08336; -. 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..1054 FT /note="Bifunctional cytochrome P450/NADPH--P450 reductase FT 2" FT /id="PRO_0000052207" FT DOMAIN 486..625 FT /note="Flavodoxin-like" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00088" FT DOMAIN 663..896 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 462..482 FT /note="Disordered" FT /evidence="ECO:0000256|SAM:MobiDB-lite" FT REGION 476..1053 FT /note="NADPH--P450 reductase" FT /evidence="ECO:0000305|PubMed:15122913" FT COMPBIAS 462..481 FT /note="Basic and acidic residues" FT /evidence="ECO:0000256|SAM:MobiDB-lite" 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 492..497 FT /ligand="FMN" FT /ligand_id="ChEBI:CHEBI:58210" FT /evidence="ECO:0000250|UniProtKB:P14779" FT BINDING 539..542 FT /ligand="FMN" FT /ligand_id="ChEBI:CHEBI:58210" FT /evidence="ECO:0000250|UniProtKB:P14779" FT BINDING 573..575 FT /ligand="FMN" FT /ligand_id="ChEBI:CHEBI:58210" FT /evidence="ECO:0000250|UniProtKB:P14779" FT BINDING 581..583 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 89 FT /note="F->V: Hydroxylates shorter substrates with higher FT conversion rates than wild-type, and in contrast to FT wild-type, is also able to convert medium-chain alkanes and FT aromatic compounds; when associated with Q-190." FT /evidence="ECO:0000269|PubMed:14741768" FT MUTAGEN 190 FT /note="S->Q: Hydroxylates shorter substrates with higher FT conversion rates than wild-type, and in contrast to FT wild-type, is also able to convert medium-chain alkanes and FT aromatic compounds; when associated with V-89." FT /evidence="ECO:0000269|PubMed:14741768" SQ SEQUENCE 1054 AA; 118676 MW; 705F8E27866CA110 CRC64; MKQASAIPQP KTYGPLKNLP HLEKEQLSQS LWRIADELGP IFRFDFPGVS SVFVSGHNLV AEVCDEKRFD KNLGKGLQKV REFGGDGLFT SWTHEPNWQK AHRILLPSFS QKAMKGYHSM MLDIATQLIQ KWSRLNPNEE IDVADDMTRL TLDTIGLCGF NYRFNSFYRD SQHPFITSML RALKEAMNQS KRLGLQDKMM VKTKLQFQKD IEVMNSLVDR MIAERKANPD ENIKDLLSLM LYAKDPVTGE TLDDENIRYQ IITFLIAGHE TTSGLLSFAI YCLLTHPEKL KKAQEEADRV LTDDTPEYKQ IQQLKYIRMV LNETLRLYPT APAFSLYAKE DTVLGGEYPI SKGQPVTVLI PKLHRDQNAW GPDAEDFRPE RFEDPSSIPH HAYKPFGNGQ RACIGMQFAL QEATMVLGLV LKHFELINHT GYELKIKEAL TIKPDDFKIT VKPRKTAAIN VQRKEQADIK AETKPKETKP KHGTPLLVLF GSNLGTAEGI AGELAAQGRQ MGFTAETAPL DDYIGKLPEE GAVVIVTASY NGAPPDNAAG FVEWLKELEE GQLKGVSYAV FGCGNRSWAS TYQRIPRLID DMMKAKGASR LTAIGEGDAA DDFESHRESW ENRFWKETMD AFDINEIAQK EDRPSLSITF LSEATETPVA KAYGAFEGIV LENRELQTAA STRSTRHIEL EIPAGKTYKE GDHIGILPKN SRELVQRVLS RFGLQSNHVI KVSGSAHMAH LPMDRPIKVV DLLSSYVELQ EPASRLQLRE LASYTVCPPH QKELEQLVSD DGIYKEQVLA KRLTMLDFLE DYPACEMPFE RFLALLPSLK PRYYSISSSP KVHANIVSMT VGVVKASAWS GRGEYRGVAS NYLAELNTGD AAACFIRTPQ SGFQMPNDPE TPMIMVGPGT GIAPFRGFIQ ARSVLKKEGS TLGEALLYFG CRRPDHDDLY REELDQAEQD GLVTIRRCYS RVENEPKGYV QHLLKQDTQK LMTLIEKGAH IYVCGDGSQM APDVERTLRL AYEAEKAASQ EESAVWLQKL QDQRRYVKDV WTGM //