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Protein

Bifunctional cytochrome P450/NADPH--P450 reductase 1

Gene

cypD

Organism
Bacillus subtilis (strain 168)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Functions as a fatty acid monooxygenase. Catalyzes hydroxylation of a range of long-chain fatty acids, with a preference for long-chain unsaturated and branched-chain fatty acids over saturated fatty acids. Hydroxylation of myristic acid occurs mainly at the omega-2 position. Also displays a NADPH-dependent reductase activity in the C-terminal domain, which allows electron transfer from NADPH to the heme iron of the cytochrome P450 N-terminal domain (PubMed:15375636, PubMed:15122913). Is also able to catalyze efficient oxidation of sodium dodecyl sulfate (SDS) (PubMed:21048857).3 Publications

Catalytic activityi

RH + [reduced NADPH--hemoprotein reductase] + O2 = ROH + [oxidized NADPH--hemoprotein reductase] + H2O.2 Publications
NADPH + n oxidized hemoprotein = NADP+ + n reduced hemoprotein.2 Publications

Cofactori

Protein has several cofactor binding sites:

Kineticsi

kcat is 430 min(-1) for stearic acid hydroxylation. kcat is 5430 min(-1) for phytanic acid hydroxylation. kcat is 6105 min(-1) for 15-methylpalmitic acid hydroxylation. kcat is 11400 min(-1) for the reduction of cytochrome c. kcat is 38150 min(-1) for the reduction of ferricyanide (PubMed:15122913). kcat is 2244 min(-1) for oleic acid hydroxylation (PubMed:15375636).2 Publications

  1. KM=38.8 µM for stearic acid1 Publication
  2. KM=150 µM for phytanic acid1 Publication
  3. KM=56.8 µM for 15-methylpalmitic acid1 Publication
  4. KM=3.6 µM for NADPH1 Publication
  5. KM=17.9 mM for NADH1 Publication
  6. KM=6.9 µM for cytochrome c (in the reductase assay)1 Publication
  7. KM=153.4 µM for ferricyanide (in the reductase assay)1 Publication
  8. KM=17.36 µM for oleic acid1 Publication

    pH dependencei

    Optimum pH is 7.0.1 Publication

    Temperature dependencei

    Optimum temperature is 51 degrees Celsius. However, enzyme stability is dramatically reduced at this temperature, incubation for 30 minutes at 31 and 49 degrees Celsius results in 61% and 17% residual activity, respectively. Incubation at 60 degrees Celsius leads to total inactivation of the enzyme.1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Metal bindingi403 – 4031Iron (heme axial ligand)By similarity

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi499 – 5035FMNPROSITE-ProRule annotation
    Nucleotide bindingi577 – 60933FMNPROSITE-ProRule annotationAdd
    BLAST

    GO - Molecular functioni

    • aromatase activity Source: UniProtKB-EC
    • fatty acid binding Source: UniProtKB
    • flavin adenine dinucleotide binding Source: UniProtKB
    • FMN binding Source: UniProtKB
    • heme binding Source: UniProtKB
    • iron ion binding Source: InterPro
    • NADPH-hemoprotein reductase activity Source: UniProtKB
    • 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 Source: UniProtKB

    GO - Biological processi

    • fatty acid oxidation Source: UniProtKB
    Complete GO annotation...

    Keywords - Molecular functioni

    Monooxygenase, Oxidoreductase

    Keywords - Biological processi

    Electron transport, Transport

    Keywords - Ligandi

    FAD, Flavoprotein, FMN, Heme, Iron, Metal-binding, NADP

    Enzyme and pathway databases

    BioCyciBSUB:BSU07250-MONOMER.
    SABIO-RKO08394.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Bifunctional cytochrome P450/NADPH--P450 reductase 1Curated
    Alternative name(s):
    CYP102A21 Publication
    Fatty acid hydroxylase CypDCurated
    Flavocytochrome P450 102A2Curated
    Including the following 2 domains:
    Cytochrome P450 102A2Curated (EC:1.14.14.12 Publications)
    NADPH--cytochrome P450 reductaseCurated (EC:1.6.2.42 Publications)
    Gene namesi
    Name:cypDCombined sources
    Synonyms:cyp102A22 Publications, yetO1 Publication, yfnJ
    Ordered Locus Names:BSU07250
    OrganismiBacillus subtilis (strain 168)
    Taxonomic identifieri224308 [NCBI]
    Taxonomic lineageiBacteriaFirmicutesBacilliBacillalesBacillaceaeBacillus
    ProteomesiUP000001570 Componenti: Chromosome

    Organism-specific databases

    GenoListiBSU07250. [Micado]

    Subcellular locationi

    GO - Cellular componenti

    Complete GO annotation...

    Keywords - Cellular componenti

    Cytoplasm

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi15 – 151P → S: Exhibits modified substrate specificity. Shows approximately 6- to 9-fold increased activity with SDS, lauric acid and 1,4-naphthoquinone, and enhanced activity for other substrates such as ethacrynic acid and epsilon-amino-n-caproic acid. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 10611061Bifunctional cytochrome P450/NADPH--P450 reductase 1PRO_0000052206Add
    BLAST

    Proteomic databases

    PaxDbiO08394.

    Interactioni

    Protein-protein interaction databases

    STRINGi224308.Bsubs1_010100004068.

    Structurei

    3D structure databases

    ProteinModelPortaliO08394.
    SMRiO08394. Positions 7-454, 476-1059.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini493 – 632140Flavodoxin-likePROSITE-ProRule annotationAdd
    BLAST
    Domaini671 – 904234FAD-binding FR-typePROSITE-ProRule annotationAdd
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni1 – 475475Cytochrome P4501 PublicationAdd
    BLAST
    Regioni476 – 1061586NADPH--P450 reductase1 PublicationAdd
    BLAST

    Sequence similaritiesi

    In the N-terminal section; belongs to the cytochrome P450 family.Curated
    Contains 1 FAD-binding FR-type domain.PROSITE-ProRule annotation
    Contains 1 flavodoxin-like domain.PROSITE-ProRule annotation

    Phylogenomic databases

    eggNOGiENOG4107EER. Bacteria.
    COG0369. LUCA.
    COG2124. LUCA.
    HOGENOMiHOG000093545.
    InParanoidiO08394.
    KOiK14338.
    OMAiCEIRFER.
    OrthoDBiEOG63Z73K.
    PhylomeDBiO08394.

    Family and domain databases

    Gene3Di1.10.630.10. 1 hit.
    1.20.990.10. 1 hit.
    3.40.50.360. 1 hit.
    InterProiIPR023206. Bifunctional_P450_P450_red.
    IPR001128. Cyt_P450.
    IPR017972. Cyt_P450_CS.
    IPR003097. FAD-binding_1.
    IPR017927. Fd_Rdtase_FAD-bd.
    IPR001094. Flavdoxin-like.
    IPR008254. Flavodoxin/NO_synth.
    IPR001709. Flavoprot_Pyr_Nucl_cyt_Rdtase.
    IPR029039. Flavoprotein-like_dom.
    IPR023173. NADPH_Cyt_P450_Rdtase_dom3.
    IPR001433. OxRdtase_FAD/NAD-bd.
    IPR017938. Riboflavin_synthase-like_b-brl.
    [Graphical view]
    PfamiPF00667. FAD_binding_1. 1 hit.
    PF00258. Flavodoxin_1. 1 hit.
    PF00175. NAD_binding_1. 1 hit.
    PF00067. p450. 1 hit.
    [Graphical view]
    PIRSFiPIRSF000209. Bifunctional_P450_P450R. 1 hit.
    PRINTSiPR00369. FLAVODOXIN.
    PR00371. FPNCR.
    SUPFAMiSSF48264. SSF48264. 1 hit.
    SSF52218. SSF52218. 1 hit.
    SSF63380. SSF63380. 1 hit.
    PROSITEiPS00086. CYTOCHROME_P450. 1 hit.
    PS51384. FAD_FR. 1 hit.
    PS50902. FLAVODOXIN_LIKE. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    O08394-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MKETSPIPQP KTFGPLGNLP LIDKDKPTLS LIKLAEEQGP IFQIHTPAGT
    60 70 80 90 100
    TIVVSGHELV KEVCDEERFD KSIEGALEKV RAFSGDGLFT SWTHEPNWRK
    110 120 130 140 150
    AHNILMPTFS QRAMKDYHEK MVDIAVQLIQ KWARLNPNEA VDVPGDMTRL
    160 170 180 190 200
    TLDTIGLCGF NYRFNSYYRE TPHPFINSMV RALDEAMHQM QRLDVQDKLM
    210 220 230 240 250
    VRTKRQFRYD IQTMFSLVDS IIAERRANGD QDEKDLLARM LNVEDPETGE
    260 270 280 290 300
    KLDDENIRFQ IITFLIAGHE TTSGLLSFAT YFLLKHPDKL KKAYEEVDRV
    310 320 330 340 350
    LTDAAPTYKQ VLELTYIRMI LNESLRLWPT APAFSLYPKE DTVIGGKFPI
    360 370 380 390 400
    TTNDRISVLI PQLHRDRDAW GKDAEEFRPE RFEHQDQVPH HAYKPFGNGQ
    410 420 430 440 450
    RACIGMQFAL HEATLVLGMI LKYFTLIDHE NYELDIKQTL TLKPGDFHIS
    460 470 480 490 500
    VQSRHQEAIH ADVQAAEKAA PDEQKEKTEA KGASVIGLNN RPLLVLYGSD
    510 520 530 540 550
    TGTAEGVARE LADTASLHGV RTKTAPLNDR IGKLPKEGAV VIVTSSYNGK
    560 570 580 590 600
    PPSNAGQFVQ WLQEIKPGEL EGVHYAVFGC GDHNWASTYQ YVPRFIDEQL
    610 620 630 640 650
    AEKGATRFSA RGEGDVSGDF EGQLDEWKKS MWADAIKAFG LELNENADKE
    660 670 680 690 700
    RSTLSLQFVR GLGESPLARS YEASHASIAE NRELQSADSD RSTRHIEIAL
    710 720 730 740 750
    PPDVEYQEGD HLGVLPKNSQ TNVSRILHRF GLKGTDQVTL SASGRSAGHL
    760 770 780 790 800
    PLGRPVSLHD LLSYSVEVQE AATRAQIREL ASFTVCPPHR RELEELSAEG
    810 820 830 840 850
    VYQEQILKKR ISMLDLLEKY EACDMPFERF LELLRPLKPR YYSISSSPRV
    860 870 880 890 900
    NPRQASITVG VVRGPAWSGR GEYRGVASND LAERQAGDDV VMFIRTPESR
    910 920 930 940 950
    FQLPKDPETP IIMVGPGTGV APFRGFLQAR DVLKREGKTL GEAHLYFGCR
    960 970 980 990 1000
    NDRDFIYRDE LERFEKDGIV TVHTAFSRKE GMPKTYVQHL MADQADTLIS
    1010 1020 1030 1040 1050
    ILDRGGRLYV CGDGSKMAPD VEAALQKAYQ AVHGTGEQEA QNWLRHLQDT
    1060
    GMYAKDVWAG I
    Length:1,061
    Mass (Da):119,468
    Last modified:July 1, 1997 - v1
    Checksum:i7915DACC20578978
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D87979 Genomic DNA. Translation: BAA20123.1.
    AL009126 Genomic DNA. Translation: CAB12544.1.
    PIRiD69799.
    RefSeqiNP_388606.1. NC_000964.3.
    WP_003242884.1. NZ_JNCM01000032.1.

    Genome annotation databases

    EnsemblBacteriaiCAB12544; CAB12544; BSU07250.
    GeneIDi938784.
    KEGGibsu:BSU07250.
    PATRICi18973100. VBIBacSub10457_0764.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D87979 Genomic DNA. Translation: BAA20123.1.
    AL009126 Genomic DNA. Translation: CAB12544.1.
    PIRiD69799.
    RefSeqiNP_388606.1. NC_000964.3.
    WP_003242884.1. NZ_JNCM01000032.1.

    3D structure databases

    ProteinModelPortaliO08394.
    SMRiO08394. Positions 7-454, 476-1059.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi224308.Bsubs1_010100004068.

    Proteomic databases

    PaxDbiO08394.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiCAB12544; CAB12544; BSU07250.
    GeneIDi938784.
    KEGGibsu:BSU07250.
    PATRICi18973100. VBIBacSub10457_0764.

    Organism-specific databases

    GenoListiBSU07250. [Micado]

    Phylogenomic databases

    eggNOGiENOG4107EER. Bacteria.
    COG0369. LUCA.
    COG2124. LUCA.
    HOGENOMiHOG000093545.
    InParanoidiO08394.
    KOiK14338.
    OMAiCEIRFER.
    OrthoDBiEOG63Z73K.
    PhylomeDBiO08394.

    Enzyme and pathway databases

    BioCyciBSUB:BSU07250-MONOMER.
    SABIO-RKO08394.

    Family and domain databases

    Gene3Di1.10.630.10. 1 hit.
    1.20.990.10. 1 hit.
    3.40.50.360. 1 hit.
    InterProiIPR023206. Bifunctional_P450_P450_red.
    IPR001128. Cyt_P450.
    IPR017972. Cyt_P450_CS.
    IPR003097. FAD-binding_1.
    IPR017927. Fd_Rdtase_FAD-bd.
    IPR001094. Flavdoxin-like.
    IPR008254. Flavodoxin/NO_synth.
    IPR001709. Flavoprot_Pyr_Nucl_cyt_Rdtase.
    IPR029039. Flavoprotein-like_dom.
    IPR023173. NADPH_Cyt_P450_Rdtase_dom3.
    IPR001433. OxRdtase_FAD/NAD-bd.
    IPR017938. Riboflavin_synthase-like_b-brl.
    [Graphical view]
    PfamiPF00667. FAD_binding_1. 1 hit.
    PF00258. Flavodoxin_1. 1 hit.
    PF00175. NAD_binding_1. 1 hit.
    PF00067. p450. 1 hit.
    [Graphical view]
    PIRSFiPIRSF000209. Bifunctional_P450_P450R. 1 hit.
    PRINTSiPR00369. FLAVODOXIN.
    PR00371. FPNCR.
    SUPFAMiSSF48264. SSF48264. 1 hit.
    SSF52218. SSF52218. 1 hit.
    SSF63380. SSF63380. 1 hit.
    PROSITEiPS00086. CYTOCHROME_P450. 1 hit.
    PS51384. FAD_FR. 1 hit.
    PS50902. FLAVODOXIN_LIKE. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Sequence of the Bacillus subtilis genome region in the vicinity of the lev operon reveals two new extracytoplasmic function RNA polymerase sigma factors SigV and SigZ."
      Sorokin A., Bolotin A., Purnelle B., Hilbert H., Lauber J., Duesterhoeft A., Ehrlich S.D.
      Microbiology 143:2939-2943(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: 168.
    2. "The complete genome sequence of the Gram-positive bacterium Bacillus subtilis."
      Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V., Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R., Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S., Bruschi C.V.
      , Caldwell B., Capuano V., Carter N.M., Choi S.-K., Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F., Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D., Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M., Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P., Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K., Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S., Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y., Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G., Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J., Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C., Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S., Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B., Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S., Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M., Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y., Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J., Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A., Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M., Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S., Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E., Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K., Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E., Yoshikawa H., Danchin A.
      Nature 390:249-256(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: 168.
    3. "Cloning, expression and characterisation of CYP102A2, a self-sufficient P450 monooxygenase from Bacillus subtilis."
      Budde M., Maurer S.C., Schmid R.D., Urlacher V.B.
      Appl. Microbiol. Biotechnol. 66:180-186(2004) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES.
    4. "Expression, purification, and characterization of Bacillus subtilis cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of P450 BM3 from Bacillus megaterium."
      Gustafsson M.C., Roitel O., Marshall K.R., Noble M.A., Chapman S.K., Pessegueiro A., Fulco A.J., Cheesman M.R., von Wachenfeldt C., Munro A.W.
      Biochemistry 43:5474-5487(2004) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES.
      Strain: 168 / 1A1.
    5. "Engineering the substrate specificity of cytochrome P450 CYP102A2 by directed evolution: production of an efficient enzyme for bioconversion of fine chemicals."
      Axarli I., Prigipaki A., Labrou N.E.
      Biomol. Eng. 22:81-88(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN ENGINEERING, MUTAGENESIS OF PRO-15.
      Strain: 168.
    6. "Cytochrome P450 102A2 catalyzes efficient oxidation of sodium dodecyl sulphate: a molecular tool for remediation."
      Axarli I., Prigipaki A., Labrou N.E.
      Enzyme Res. 2010:125429-125429(2010) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION.

    Entry informationi

    Entry nameiCYPD_BACSU
    AccessioniPrimary (citable) accession number: O08394
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: December 1, 2000
    Last sequence update: July 1, 1997
    Last modified: January 20, 2016
    This is version 128 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    Complete proteome, Multifunctional enzyme, Reference proteome

    Documents

    1. Bacillus subtilis
      Bacillus subtilis (strain 168): entries, gene names and cross-references to SubtiList
    2. SIMILARITY comments
      Index of protein domains and families

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.