CYPB_BACSU
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 03-AUG-2022, entry version 162.
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 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}.
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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; O08336; -.
DR PRIDE; O08336; -.
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 OMA; EDYPACE; -.
DR PhylomeDB; O08336; -.
DR BioCyc; BSUB:BSU27160-MON; -.
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 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..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 wild-
FT 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 wild-
FT 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
