ID CRTM_STAA8 Reviewed; 287 AA. AC Q2FV59; DT 03-APR-2007, integrated into UniProtKB/Swiss-Prot. DT 21-MAR-2006, sequence version 1. DT 07-OCT-2020, entry version 88. DE RecName: Full=4,4'-diapophytoene synthase {ECO:0000303|PubMed:16269684}; DE Short=DAP synthase {ECO:0000303|PubMed:16269684}; DE EC=2.5.1.96 {ECO:0000269|PubMed:12426357, ECO:0000269|PubMed:16269684}; DE AltName: Full=C30 carotenoid synthase {ECO:0000303|PubMed:12426357}; DE AltName: Full=Dehydrosqualene synthase {ECO:0000303|PubMed:12426357}; GN Name=crtM {ECO:0000303|PubMed:12426357}; OrderedLocusNames=SAOUHSC_02879; OS Staphylococcus aureus (strain NCTC 8325 / PS 47). OC Bacteria; Firmicutes; Bacilli; Bacillales; Staphylococcaceae; OC Staphylococcus. OX NCBI_TaxID=93061; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=NCTC 8325 / PS 47; RA Gillaspy A.F., Worrell V., Orvis J., Roe B.A., Dyer D.W., Iandolo J.J.; RT "The Staphylococcus aureus NCTC 8325 genome."; RL (In) Fischetti V., Novick R., Ferretti J., Portnoy D., Rood J. (eds.); RL Gram positive pathogens, 2nd edition, pp.381-412, ASM Press, Washington RL D.C. (2006). RN [2] RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND MUTAGENESIS OF RP PHE-26. RX PubMed=12426357; DOI=10.1128/jb.184.23.6690-6699.2002; RA Umeno D., Tobias A.V., Arnold F.H.; RT "Evolution of the C(30) carotenoid synthase crtM for function in a C(40) RT pathway."; RL J. Bacteriol. 184:6690-6699(2002). RN [3] RP SUBSTRATE SPECIFICITY. RX PubMed=12788765; DOI=10.1128/aem.69.6.3573-3579.2003; RA Umeno D., Arnold F.H.; RT "A C(35) carotenoid biosynthetic pathway."; RL Appl. Environ. Microbiol. 69:3573-3579(2003). RN [4] RP MUTAGENESIS OF TRP-38 AND GLU-180. RX PubMed=12907743; DOI=10.1093/nar/gng091; RA Umeno D., Hiraga K., Arnold F.H.; RT "Method to protect a targeted amino acid residue during random RT mutagenesis."; RL Nucleic Acids Res. 31:E91-E91(2003). RN [5] RP MUTAGENESIS OF PHE-26; TRP-38 AND GLU-180. RX PubMed=14973014; DOI=10.1128/jb.186.5.1531-1536.2004; RA Umeno D., Arnold F.H.; RT "Evolution of a pathway to novel long-chain carotenoids."; RL J. Bacteriol. 186:1531-1536(2004). RN [6] RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES. RX PubMed=16269684; DOI=10.1128/aem.71.11.6578-6583.2005; RA Ku B., Jeong J.-C., Mijts B.N., Schmidt-Dannert C., Dordick J.S.; RT "Preparation, characterization, and optimization of an in vitro C(30) RT carotenoid pathway."; RL Appl. Environ. Microbiol. 71:6578-6583(2005). RN [7] RP FUNCTION, AND PATHWAY. RC STRAIN=KCTC 1928; RX PubMed=22535955; DOI=10.1074/jbc.m112.343020; RA Kim S.H., Lee P.C.; RT "Functional expression and extension of staphylococcal staphyloxanthin RT biosynthetic pathway in Escherichia coli."; RL J. Biol. Chem. 287:21575-21583(2012). CC -!- FUNCTION: Involved in the biosynthesis of the yellow-orange carotenoid CC staphyloxanthin, which plays a role in the virulence via its protective CC function against oxidative stress. Catalyzes the head-to-head CC condensation of two molecules of farnesyl diphosphate (FPP) into the CC colorless C(30) carotenoid 4,4'-diapophytoene (dehydrosqualene). CC {ECO:0000269|PubMed:12426357, ECO:0000269|PubMed:16269684, CC ECO:0000269|PubMed:22535955}. CC -!- CATALYTIC ACTIVITY: CC Reaction=2 (2E,6E)-farnesyl diphosphate = 15-cis-4,4'-diapophytoene + 2 CC diphosphate; Xref=Rhea:RHEA:31547, ChEBI:CHEBI:33019, CC ChEBI:CHEBI:62738, ChEBI:CHEBI:175763; EC=2.5.1.96; CC Evidence={ECO:0000269|PubMed:12426357, ECO:0000269|PubMed:16269684}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000250|UniProtKB:A9JQL9}; CC Note=Binds 2 Mg(2+) ions per subunit. {ECO:0000250|UniProtKB:A9JQL9}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=1.2 uM for farnesyl diphosphate {ECO:0000269|PubMed:16269684}; CC Vmax=176 uM/h/mg enzyme {ECO:0000269|PubMed:16269684}; CC -!- PATHWAY: Carotenoid biosynthesis; staphyloxanthin biosynthesis; CC staphyloxanthin from farnesyl diphosphate: step 1/5. CC {ECO:0000269|PubMed:22535955}. CC -!- MISCELLANEOUS: CrtM is not functional in a C(40) pathway, however CC independent mutations on Phe-26, Trp-38 or Glu-180 are sufficient to CC permit the synthesis of C(40) carotenoids, such as lycopene and CC 3,4,3',4'-tetrahydrolycopene, although there is a decrease in the CC synthesis of C(30) compounds. The combination of mutations at Phe-26 CC and Trp-38 appears to be harmful for the general performance of the CC enzyme. {ECO:0000269|PubMed:12426357}. CC -!- MISCELLANEOUS: Upon coexpression with Erwinia geranylgeranyldiphosphate CC (GGDP) synthase, CrtM produces novel carotenoids with the asymmetrical CC C(35) backbone, such as 4-apophytoene, and the production of the CC natural product 4,4'-diapophytoene drops dramatically. CC {ECO:0000269|PubMed:12788765}. CC -!- SIMILARITY: Belongs to the phytoene/squalene synthase family. CrtM CC subfamily. {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; CP000253; ABD31876.1; -; Genomic_DNA. DR RefSeq; WP_000178307.1; NZ_LS483365.1. DR RefSeq; YP_501333.1; NC_007795.1. DR SMR; Q2FV59; -. DR STRING; 1280.SAXN108_2813; -. DR EnsemblBacteria; ABD31876; ABD31876; SAOUHSC_02879. DR GeneID; 3921549; -. DR GeneID; 45575712; -. DR KEGG; sao:SAOUHSC_02879; -. DR PATRIC; fig|93061.5.peg.2602; -. DR eggNOG; COG1562; Bacteria. DR HOGENOM; CLU_037269_1_3_9; -. DR KO; K10208; -. DR OMA; KRRAIWA; -. DR SABIO-RK; Q2FV59; -. DR UniPathway; UPA00029; UER00556. DR PRO; PR:Q2FV59; -. DR Proteomes; UP000008816; Chromosome. DR GO; GO:0004310; F:farnesyl-diphosphate farnesyltransferase activity; IEA:InterPro. DR GO; GO:0016767; F:geranylgeranyl-diphosphate geranylgeranyltransferase activity; IBA:GO_Central. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0051996; F:squalene synthase activity; IEA:InterPro. DR GO; GO:0016117; P:carotenoid biosynthetic process; IBA:GO_Central. DR GO; GO:0009405; P:pathogenesis; IEA:UniProtKB-KW. DR CDD; cd00683; Trans_IPPS_HH; 1. DR Gene3D; 1.10.600.10; -; 1. DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf. DR InterPro; IPR002060; Squ/phyt_synthse. DR InterPro; IPR019845; Squalene/phytoene_synthase_CS. DR InterPro; IPR033904; Trans_IPPS_HH. DR SFLD; SFLDG01018; Squalene/Phytoene_Synthase_Lik; 1. DR SUPFAM; SSF48576; SSF48576; 1. DR PROSITE; PS01044; SQUALEN_PHYTOEN_SYN_1; 1. PE 1: Evidence at protein level; KW Carotenoid biosynthesis; Magnesium; Metal-binding; Reference proteome; KW Transferase; Virulence. FT CHAIN 1..287 FT /note="4,4'-diapophytoene synthase" FT /id="PRO_0000282616" FT REGION 18..21 FT /note="Farnesyl diphosphate 1 binding" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT METAL 48 FT /note="Magnesium 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT METAL 52 FT /note="Magnesium 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT METAL 168 FT /note="Magnesium 2" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT METAL 172 FT /note="Magnesium 2" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 41 FT /note="Farnesyl diphosphate 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 45 FT /note="Farnesyl diphosphate 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 45 FT /note="Farnesyl diphosphate 2" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 165 FT /note="Farnesyl diphosphate 2" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 171 FT /note="Farnesyl diphosphate 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT BINDING 248 FT /note="Farnesyl diphosphate 1" FT /evidence="ECO:0000250|UniProtKB:A9JQL9" FT MUTAGEN 26 FT /note="F->A,G,L,S: Decrease in C(30) carotene synthase FT activity. C(40) carotene synthase activity acquired." FT /evidence="ECO:0000269|PubMed:12426357, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 26 FT /note="F->A: Decrease in C(30) and C(40) carotene synthase FT activities; when associated with A-38 or G-38." FT /evidence="ECO:0000269|PubMed:12426357, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 26 FT /note="F->G: Decrease in C(30) and C(40) carotene synthase FT activities; when associated with A-38 or G-38." FT /evidence="ECO:0000269|PubMed:12426357, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 38 FT /note="W->A: Decrease in C(30) and C(40) carotene synthase FT activities; when associated with A-26 or G-26." FT /evidence="ECO:0000269|PubMed:12907743, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 38 FT /note="W->C: Decrease in C(30) carotene synthase activity. FT C(40) carotene synthase activity acquired." FT /evidence="ECO:0000269|PubMed:12907743, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 38 FT /note="W->G: Decrease in C(30) and C(40) carotene synthase FT activities; when associated with A-26 or G-26." FT /evidence="ECO:0000269|PubMed:12907743, FT ECO:0000269|PubMed:14973014" FT MUTAGEN 180 FT /note="E->G: Slight increase in C(30) carotene synthase FT activity. C(40) carotene synthase activity acquired." FT /evidence="ECO:0000269|PubMed:12907743, FT ECO:0000269|PubMed:14973014" SQ SEQUENCE 287 AA; 34231 MW; 2407009413D42E63 CRC64; MTMMDMNFKY CHKIMKKHSK SFSYAFDLLP EDQRKAVWAI YAVCRKIDDS IDVYGDIQFL NQIKEDIQSI EKYPYEHHHF QSDRRIMMAL QHVAQHKNIA FQSFYNLIDT VYKDQHFTMF ETDAELFGYC YGVAGTVGEV LTPILSDHET HQTYDVARRL GESLQLINIL RDVGEDFDNE RIYFSKQRLK QYEVDIAEVY QNGVNNHYID LWEYYAAIAE KDFQDVMDQI KVFSIEAQPI IELAARIYIE ILDEVRQANY TLHERVFVDK RKKAKLFHEI NSKYHRI //