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 25-OCT-2017, entry version 139. 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; 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 RT the lev operon reveals two new extracytoplasmic function RNA RT polymerase sigma 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., RA Azevedo V., Bertero M.G., Bessieres P., Bolotin A., Borchert S., RA Borriss R., Boursier L., Brans A., Braun M., Brignell S.C., Bron S., RA Brouillet S., Bruschi C.V., Caldwell B., Capuano V., Carter N.M., RA Choi S.-K., Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., RA Denizot F., Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., RA Entian K.-D., Errington J., Fabret C., Ferrari E., Foulger D., RA Fritz C., Fujita M., Fujita Y., Fuma S., Galizzi A., Galleron N., RA Ghim S.-Y., Glaser P., Goffeau A., Golightly E.J., Grandi G., RA Guiseppi G., Guy B.J., Haga K., Haiech J., Harwood C.R., Henaut A., RA Hilbert H., Holsappel S., Hosono S., Hullo M.-F., Itaya M., RA Jones L.-M., Joris B., Karamata D., Kasahara Y., Klaerr-Blanchard M., RA Klein C., Kobayashi Y., Koetter P., Koningstein G., Krogh S., RA 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., RA Rey M., Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., RA Rose M., Sadaie Y., Sato T., Scanlan E., Schleich S., Schroeter R., RA Scoffone F., Sekiguchi J., Sekowska A., Seror S.J., Serror P., RA Shin B.-S., Soldo B., Sorokin A., Tacconi E., Takagi T., Takahashi H., RA Takemaru K., Takeuchi M., Tamakoshi A., Tanaka T., Terpstra P., RA Tognoni A., Tosato V., Uchiyama S., Vandenbol M., Vannier F., RA Vassarotti A., Viari A., Wambutt R., Wedler E., Wedler H., RA 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 RT operon by the BscR repressor and differential induction of cytochrome RT CYP102A3 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 RT of 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., RA Munro A.W.; RT "Expression, purification, and characterization of Bacillus subtilis RT cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of RT P450 BM3 from Bacillus megaterium."; RL Biochemistry 43:5474-5487(2004). RN [6] RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND MUTAGENESIS RP OF PHE-89 AND SER-190. RC STRAIN=168 / ATCC 33234 / DSM 402 / 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 RT (CYP102A3) from Bacillus subtilis."; RL J. Biotechnol. 108:41-49(2004). RN [7] RP PROTEIN ENGINEERING. RC STRAIN=168 / ATCC 33234 / DSM 402 / NCIMB 10106; RX PubMed=16381045; DOI=10.1002/cbic.200500266; RA Lentz O., Feenstra A., Habicher T., Hauer B., Schmid R.D., RA 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 CC a preference for long-chain unsaturated and branched-chain fatty CC acids over saturated fatty acids. Hydroxylation of myristic acid CC occurs mainly at the omega-2 and omega-3 positions, in CC approximately equal proportions. Also displays a NADPH-dependent CC reductase activity in the C-terminal domain, which allows electron CC transfer from NADPH to the heme iron of the cytochrome P450 N- CC terminal domain. {ECO:0000269|PubMed:14741768, CC ECO:0000269|PubMed:15122913}. CC -!- CATALYTIC ACTIVITY: RH + [reduced NADPH--hemoprotein reductase] + CC O(2) = ROH + [oxidized NADPH--hemoprotein reductase] + H(2)O. CC {ECO:0000269|PubMed:14741768, ECO:0000269|PubMed:15122913}. CC -!- CATALYTIC ACTIVITY: NADPH + n oxidized hemoprotein = NADP(+) + n CC reduced hemoprotein. {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 CC {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 CC 5556 min(-1) for myristic acid hydroxylation. kcat is 676 min(- CC 1) for palmitic acid hydroxylation. kcat is 374 min(-1) for CC stearic acid hydroxylation. kcat is 794 min(-1) for phytanic CC acid hydroxylation. kcat is 3845 min(-1) for 15-methylpalmitic CC acid hydroxylation. kcat is 1690 min(-1) for arachidonic acid CC hydroxylation. kcat is 3520 min(-1) for the reduction of CC cytochrome c. kcat is 37050 min(-1) for the reduction of CC ferricyanide. {ECO:0000269|PubMed:15122913}; CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}. CC -!- INDUCTION: Negatively regulated by the transcriptional repressor CC FatR (PubMed:11574077, PubMed:11734890). Is induced by fatty acids CC such as oleate, linoleate and phytanate, that bind and displace CC the FatR repressor (PubMed:11734890). Is also induced by CC palmitate, likely via another mechanism (PubMed:11574077). CC Transcribed under partial control of SigM ECF sigma factor CC (PubMed:17434969). {ECO:0000269|PubMed:11574077, CC ECO:0000269|PubMed:11734890, ECO:0000269|PubMed:17434969}. CC -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome CC 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; 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 ProteinModelPortal; O08336; -. DR SMR; O08336; -. DR STRING; 224308.Bsubs1_010100014841; -. DR PaxDb; O08336; -. DR EnsemblBacteria; CAB14658; CAB14658; BSU27160. DR GeneID; 937585; -. DR KEGG; bsu:BSU27160; -. DR PATRIC; fig|224308.179.peg.2949; -. DR eggNOG; ENOG4107EER; Bacteria. DR eggNOG; COG0369; LUCA. DR HOGENOM; HOG000093545; -. DR InParanoid; O08336; -. DR KO; K14338; -. DR OMA; EDYPACE; -. DR PhylomeDB; O08336; -. DR BioCyc; BSUB:BSU27160-MONOMER; -. DR SABIO-RK; O08336; -. DR Proteomes; UP000001570; Chromosome. DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell. 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: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; 3.40.50.360; -; 1. DR InterPro; IPR023206; Bifunctional_P450_P450_red. DR InterPro; IPR001128; Cyt_P450. DR InterPro; IPR017972; Cyt_P450_CS. DR InterPro; IPR036396; Cyt_P450_sf. DR InterPro; IPR003097; FAD-binding_1. DR InterPro; IPR017927; Fd_Rdtase_FAD-bd. DR InterPro; IPR001094; Flavdoxin-like. DR InterPro; IPR008254; Flavodoxin/NO_synth. DR InterPro; IPR001709; Flavoprot_Pyr_Nucl_cyt_Rdtase. DR InterPro; IPR029039; Flavoprotein-like_dom. 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; 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 Complete proteome; Cytoplasm; Electron transport; FAD; Flavoprotein; KW FMN; Heme; Iron; Metal-binding; Monooxygenase; Multifunctional enzyme; KW NADP; Oxidoreductase; Reference proteome; Transport. FT CHAIN 1 1054 Bifunctional cytochrome P450/NADPH--P450 FT reductase 2. FT /FTId=PRO_0000052207. FT DOMAIN 486 625 Flavodoxin-like. {ECO:0000255|PROSITE- FT ProRule:PRU00088}. FT DOMAIN 663 896 FAD-binding FR-type. FT {ECO:0000255|PROSITE-ProRule:PRU00716}. FT NP_BIND 492 497 FMN. {ECO:0000250|UniProtKB:P14779}. FT NP_BIND 539 542 FMN. {ECO:0000250|UniProtKB:P14779}. FT NP_BIND 573 575 FMN. {ECO:0000250|UniProtKB:P14779}. FT NP_BIND 581 583 FMN. {ECO:0000250|UniProtKB:P14779}. FT REGION 1 475 Cytochrome P450. FT {ECO:0000305|PubMed:15122913}. FT REGION 22 31 Fatty acid binding. FT {ECO:0000250|UniProtKB:P14779}. FT REGION 77 89 Fatty acid binding. FT {ECO:0000250|UniProtKB:P14779}. FT REGION 331 333 Fatty acid binding. FT {ECO:0000250|UniProtKB:P14779}. FT REGION 476 1053 NADPH--P450 reductase. FT {ECO:0000305|PubMed:15122913}. FT METAL 403 403 Iron (heme axial ligand). FT {ECO:0000250|UniProtKB:P14779}. FT BINDING 183 183 Fatty acid. FT {ECO:0000250|UniProtKB:P14779}. FT BINDING 266 266 Fatty acid. FT {ECO:0000250|UniProtKB:P14779}. FT BINDING 440 440 Fatty acid. FT {ECO:0000250|UniProtKB:P14779}. FT SITE 271 271 Important for catalytic activity. FT {ECO:0000250|UniProtKB:P14779}. FT MUTAGEN 89 89 F->V: Hydroxylates shorter substrates FT with higher conversion rates than wild- FT type, and in contrast to wild-type, is FT also able to convert medium-chain alkanes FT and aromatic compounds; when associated FT with Q-190. FT {ECO:0000269|PubMed:14741768}. FT MUTAGEN 190 190 S->Q: Hydroxylates shorter substrates FT with higher conversion rates than wild- FT type, and in contrast to wild-type, is FT also able to convert medium-chain alkanes FT and aromatic compounds; when associated FT with V-89. {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 //