ID BIOB_ECOLI Reviewed; 346 AA. AC P12996; Q2MBJ4; DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot. DT 01-NOV-1997, sequence version 2. DT 27-MAR-2024, entry version 208. DE RecName: Full=Biotin synthase; DE EC=2.8.1.6 {ECO:0000269|PubMed:11862544, ECO:0000269|PubMed:17014080, ECO:0000269|PubMed:8142361}; GN Name=bioB; OrderedLocusNames=b0775, JW0758; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RX PubMed=3058702; DOI=10.1016/s0021-9258(19)77675-3; RA Otsuka A.J., Buoncristiani M.R., Howard P.K., Flamm J., Johnson O., RA Yamamoto R., Uchida K., Cook C., Ruppert J., Matsuzaki J.; RT "The Escherichia coli biotin biosynthetic enzyme sequences predicted from RT the nucleotide sequence of the bio operon."; RL J. Biol. Chem. 263:19577-19585(1988). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RA Pearson B.M., McKee R.A.; RT "Genetic material for expression of biotin synthetase enzymes."; RL Patent number GB2216530, 11-OCT-1989. RN [3] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=9278503; DOI=10.1126/science.277.5331.1453; RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., RA Shao Y.; RT "The complete genome sequence of Escherichia coli K-12."; RL Science 277:1453-1462(1997). RN [4] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=16738553; DOI=10.1038/msb4100049; RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.; RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 RT and W3110."; RL Mol. Syst. Biol. 2:E1-E5(2006). RN [5] RP PROTEIN SEQUENCE OF 1-17, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL RP PROPERTIES, COFACTOR, AND SUBUNIT. RX PubMed=8142361; DOI=10.1021/bi00178a020; RA Sanyal I., Cohen G., Flint D.H.; RT "Biotin synthase: purification, characterization as a [2Fe-2S] cluster RT protein, and in vitro activity of the Escherichia coli bioB gene product."; RL Biochemistry 33:3625-3631(1994). RN [6] RP IDENTIFICATION BY 2D-GEL. RX PubMed=9298644; DOI=10.1002/elps.1150180805; RA VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.; RT "Escherichia coli proteome analysis using the gene-protein database."; RL Electrophoresis 18:1243-1251(1997). RN [7] RP MUTAGENESIS OF CYSTEINE RESIDUES. RX PubMed=11686925; DOI=10.1093/oxfordjournals.jbchem.a003028; RA Farh L., Hwang S.-Y., Steinrauf L., Chiang H.-J., Shiuan D.; RT "Structure-function studies of Escherichia coli biotin synthase via a RT chemical modification and site-directed mutagenesis approach."; RL J. Biochem. 130:627-635(2001). RN [8] RP RESONANCE RAMAN SPECTROSCOPY, EPR SPECTROSCOPY, AND MOSSBAUER SPECTROSCOPY. RX PubMed=12440894; DOI=10.1021/ja0283044; RA Cosper M.M., Jameson G.N.L., Davydov R., Eidsness M.K., Hoffman B.M., RA Huynh B.H., Johnson M.K.; RT "The [4Fe-4S]2+ cluster in reconstituted biotin synthase binds S-adenosyl- RT L-methionine."; RL J. Am. Chem. Soc. 124:14006-14007(2002). RN [9] RP MUTAGENESIS OF CYS-53; CYS-57; CYS-60; CYS-97; CYS-128 AND CYS-188, RP CATALYTIC ACTIVITY, EPR SPECTROSCOPY, AND MOSSBAUER SPECTROSCOPY. RX PubMed=11834738; DOI=10.1074/jbc.m111324200; RA Ollagnier-de Choudens S., Sanakis Y., Hewitson K.S., Roach P., Muenck E., RA Fontecave M.; RT "Reductive cleavage of S-adenosylmethionine by biotin synthase from RT Escherichia coli."; RL J. Biol. Chem. 277:13449-13454(2002). RN [10] RP MUTAGENESIS OF CYS-53; CYS-57; CYS-60; CYS-97; CYS-128 AND CYS-188, EPR RP SPECTROSCOPY, AND MOSSBAUER SPECTROSCOPY. RX PubMed=11862544; DOI=10.1007/s007750100268; RA Hewitson K.S., Ollagnier-de Choudens S., Sanakis Y., Shaw N.M., RA Baldwin J.E., Muenck E., Roach P.L., Fontecave M.; RT "The iron-sulfur center of biotin synthase: site-directed mutants."; RL J. Biol. Inorg. Chem. 7:83-93(2002). RN [11] RP ABSORPTION SPECTROSCOPY. RX PubMed=12824504; DOI=10.1110/ps.0302203; RA Cosper M.M., Cosper N.J., Hong W., Shokes J.E., Broderick W.E., RA Broderick J.B., Johnson M.K., Scott R.A.; RT "Structural studies of the interaction of S-adenosylmethionine with the RT [4Fe-4S] clusters in biotin synthase and pyruvate formate-lyase activating RT enzyme."; RL Protein Sci. 12:1573-1577(2003). RN [12] RP ACTIVITY REGULATION. RX PubMed=15911379; DOI=10.1016/j.chembiol.2005.04.012; RA Choi-Rhee E., Cronan J.E.; RT "A nucleosidase required for in vivo function of the S-adenosyl-L- RT methionine radical enzyme, biotin synthase."; RL Chem. Biol. 12:589-593(2005). RN [13] RP REACTION MECHANISM. RX PubMed=19245793; DOI=10.1016/j.bbrc.2009.02.089; RA Lotierzo M., Bui B.T., Leech H.K., Warren M.J., Marquet A., Rigby S.E.; RT "Iron-sulfur cluster dynamics in biotin synthase: a new [2Fe-2S](1+) RT cluster."; RL Biochem. Biophys. Res. Commun. 381:487-490(2009). RN [14] RP MUTAGENESIS OF ASN-151; HIS-152; ASN-153 AND ASP-155, AND CATALYTIC RP ACTIVITY. RX PubMed=17014080; DOI=10.1021/bi060662m; RA Lotierzo M., Raux E., Tse Sum Bui B., Goasdoue N., Libot F., Florentin D., RA Warren M.J., Marquet A.; RT "Biotin synthase mechanism: mutagenesis of the YNHNLD conserved motif."; RL Biochemistry 45:12274-12281(2006). RN [15] RP X-RAY CRYSTALLOGRAPHY (3.4 ANGSTROMS). RX PubMed=14704425; DOI=10.1126/science.1088493; RA Berkovitch F., Nicolet Y., Wan J.T., Jarrett J.T., Drennan C.L.; RT "Crystal structure of biotin synthase, an S-adenosylmethionine-dependent RT radical enzyme."; RL Science 303:76-79(2004). CC -!- FUNCTION: Catalyzes the conversion of dethiobiotin (DTB) to biotin by CC the insertion of a sulfur atom into dethiobiotin via a radical-based CC mechanism. {ECO:0000269|PubMed:8142361}. CC -!- CATALYTIC ACTIVITY: CC Reaction=(4R,5S)-dethiobiotin + [sulfur carrier]-SH + 2 reduced [2Fe- CC 2S]-[ferredoxin] + 2 S-adenosyl-L-methionine = 2 5'-deoxyadenosine + CC [sulfur carrier]-H + biotin + 2 L-methionine + 2 oxidized [2Fe-2S]- CC [ferredoxin]; Xref=Rhea:RHEA:22060, Rhea:RHEA-COMP:10000, Rhea:RHEA- CC COMP:10001, Rhea:RHEA-COMP:14737, Rhea:RHEA-COMP:14739, CC ChEBI:CHEBI:17319, ChEBI:CHEBI:29917, ChEBI:CHEBI:33737, CC ChEBI:CHEBI:33738, ChEBI:CHEBI:57586, ChEBI:CHEBI:57844, CC ChEBI:CHEBI:59789, ChEBI:CHEBI:64428, ChEBI:CHEBI:149473; EC=2.8.1.6; CC Evidence={ECO:0000269|PubMed:11862544, ECO:0000269|PubMed:17014080, CC ECO:0000269|PubMed:8142361}; CC -!- COFACTOR: CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883; CC Evidence={ECO:0000269|PubMed:8142361}; CC Note=Binds 1 [4Fe-4S] cluster. The cluster is coordinated with 3 CC cysteines and an exchangeable S-adenosyl-L-methionine. CC {ECO:0000269|PubMed:8142361}; CC -!- COFACTOR: CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135; CC Evidence={ECO:0000269|PubMed:8142361}; CC Note=Binds 1 [2Fe-2S] cluster. The cluster is coordinated with 3 CC cysteines and 1 arginine. {ECO:0000269|PubMed:8142361}; CC -!- ACTIVITY REGULATION: Is physiologically inhibited by accumulation of CC the reaction product 5'-deoxyadenosine. {ECO:0000269|PubMed:15911379}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=2 uM for dethiobiotin {ECO:0000269|PubMed:8142361}; CC -!- PATHWAY: Cofactor biosynthesis; biotin biosynthesis; biotin from 7,8- CC diaminononanoate: step 2/2. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:8142361}. CC -!- INTERACTION: CC P12996; Q47147: yafJ; NbExp=4; IntAct=EBI-557917, EBI-1114805; CC -!- SIMILARITY: Belongs to the radical SAM superfamily. Biotin synthase 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; J04423; AAA23515.1; -; Genomic_DNA. DR EMBL; A11530; CAA00965.1; -; Unassigned_DNA. DR EMBL; U00096; AAC73862.1; -; Genomic_DNA. DR EMBL; AP009048; BAE76362.1; -; Genomic_DNA. DR PIR; JC2517; SYECBB. DR RefSeq; NP_415296.1; NC_000913.3. DR RefSeq; WP_000951213.1; NZ_SSZK01000002.1. DR PDB; 1R30; X-ray; 3.40 A; A/B=2-346. DR PDBsum; 1R30; -. DR AlphaFoldDB; P12996; -. DR SMR; P12996; -. DR BioGRID; 4259949; 37. DR BioGRID; 849746; 13. DR DIP; DIP-9220N; -. DR IntAct; P12996; 17. DR STRING; 511145.b0775; -. DR DrugBank; DB03775; Dethiobiotin. DR PaxDb; 511145-b0775; -. DR EnsemblBacteria; AAC73862; AAC73862; b0775. DR GeneID; 75204890; -. DR GeneID; 945370; -. DR KEGG; ecj:JW0758; -. DR KEGG; eco:b0775; -. DR PATRIC; fig|1411691.4.peg.1503; -. DR EchoBASE; EB0116; -. DR eggNOG; COG0502; Bacteria. DR HOGENOM; CLU_033172_1_2_6; -. DR InParanoid; P12996; -. DR OMA; NICTTHT; -. DR OrthoDB; 9786826at2; -. DR PhylomeDB; P12996; -. DR BioCyc; EcoCyc:BIOTIN-SYN-MONOMER; -. DR BioCyc; MetaCyc:BIOTIN-SYN-MONOMER; -. DR BRENDA; 2.8.1.6; 2026. DR SABIO-RK; P12996; -. DR UniPathway; UPA00078; UER00162. DR EvolutionaryTrace; P12996; -. DR PRO; PR:P12996; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IDA:EcoCyc. DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:EcoCyc. DR GO; GO:0004076; F:biotin synthase activity; IDA:EcoCyc. DR GO; GO:0005506; F:iron ion binding; IEA:UniProtKB-UniRule. DR GO; GO:0042803; F:protein homodimerization activity; IDA:EcoCyc. DR GO; GO:0009102; P:biotin biosynthetic process; IMP:EcoCyc. DR CDD; cd01335; Radical_SAM; 1. DR Gene3D; 3.20.20.70; Aldolase class I; 1. DR HAMAP; MF_01694; BioB; 1. DR InterPro; IPR013785; Aldolase_TIM. DR InterPro; IPR010722; BATS_dom. DR InterPro; IPR002684; Biotin_synth/BioAB. DR InterPro; IPR024177; Biotin_synthase. DR InterPro; IPR006638; Elp3/MiaA/NifB-like_rSAM. DR InterPro; IPR007197; rSAM. DR NCBIfam; TIGR00433; bioB; 1. DR PANTHER; PTHR22976; BIOTIN SYNTHASE; 1. DR PANTHER; PTHR22976:SF2; BIOTIN SYNTHASE, MITOCHONDRIAL; 1. DR Pfam; PF06968; BATS; 1. DR Pfam; PF04055; Radical_SAM; 1. DR PIRSF; PIRSF001619; Biotin_synth; 1. DR SFLD; SFLDG01060; BATS_domain_containing; 1. DR SFLD; SFLDF00272; biotin_synthase; 1. DR SMART; SM00876; BATS; 1. DR SMART; SM00729; Elp3; 1. DR SUPFAM; SSF102114; Radical SAM enzymes; 1. DR PROSITE; PS51918; RADICAL_SAM; 1. PE 1: Evidence at protein level; KW 2Fe-2S; 3D-structure; 4Fe-4S; Biotin biosynthesis; KW Direct protein sequencing; Iron; Iron-sulfur; Metal-binding; KW Reference proteome; S-adenosyl-L-methionine; Transferase. FT CHAIN 1..346 FT /note="Biotin synthase" FT /id="PRO_0000185552" FT DOMAIN 38..256 FT /note="Radical SAM core" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266" FT BINDING 53 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_note="4Fe-4S-S-AdoMet" FT BINDING 57 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_note="4Fe-4S-S-AdoMet" FT BINDING 60 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_note="4Fe-4S-S-AdoMet" FT BINDING 97 FT /ligand="[2Fe-2S] cluster" FT /ligand_id="ChEBI:CHEBI:190135" FT BINDING 128 FT /ligand="[2Fe-2S] cluster" FT /ligand_id="ChEBI:CHEBI:190135" FT BINDING 188 FT /ligand="[2Fe-2S] cluster" FT /ligand_id="ChEBI:CHEBI:190135" FT BINDING 260 FT /ligand="[2Fe-2S] cluster" FT /ligand_id="ChEBI:CHEBI:190135" FT MUTAGEN 53 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT MUTAGEN 57 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT MUTAGEN 60 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT MUTAGEN 97 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT MUTAGEN 128 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT MUTAGEN 151 FT /note="N->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:17014080" FT MUTAGEN 152 FT /note="H->A: Weak activity." FT /evidence="ECO:0000269|PubMed:17014080" FT MUTAGEN 153 FT /note="N->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:17014080" FT MUTAGEN 155 FT /note="D->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:17014080" FT MUTAGEN 188 FT /note="C->A: Total loss of activity." FT /evidence="ECO:0000269|PubMed:11834738, FT ECO:0000269|PubMed:11862544" FT CONFLICT 63 FT /note="S -> T (in Ref. 1; AAA23515)" FT /evidence="ECO:0000305" FT TURN 10..12 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 13..17 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 20..34 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 41..49 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 53..55 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 78..90 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 94..100 FT /evidence="ECO:0007829|PDB:1R30" FT TURN 107..109 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 110..122 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 125..130 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 136..145 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 149..151 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 158..164 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 170..184 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 186..188 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 191..193 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 199..210 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 211..214 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 217..223 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 240..253 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 257..264 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 265..267 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 270..279 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 283..285 FT /evidence="ECO:0007829|PDB:1R30" FT STRAND 287..294 FT /evidence="ECO:0007829|PDB:1R30" FT HELIX 298..307 FT /evidence="ECO:0007829|PDB:1R30" SQ SEQUENCE 346 AA; 38648 MW; 550A7899A2DF6082 CRC64; MAHRPRWTLS QVTELFEKPL LDLLFEAQQV HRQHFDPRQV QVSTLLSIKT GACPEDCKYC PQSSRYKTGL EAERLMEVEQ VLESARKAKA AGSTRFCMGA AWKNPHERDM PYLEQMVQGV KAMGLEACMT LGTLSESQAQ RLANAGLDYY NHNLDTSPEF YGNIITTRTY QERLDTLEKV RDAGIKVCSG GIVGLGETVK DRAGLLLQLA NLPTPPESVP INMLVKVKGT PLADNDDVDA FDFIRTIAVA RIMMPTSYVR LSAGREQMNE QTQAMCFMAG ANSIFYGCKL LTTPNPEEDK DLQLFRKLGL NPQQTAVLAG DNEQQQRLEQ ALMTPDTDEY YNAAAL //