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   28-FEB-2018, entry version 73.
DE   RecName: Full=Dehydrosqualene synthase;
DE            EC=2.5.1.96 {ECO:0000269|PubMed:12426357, ECO:0000269|PubMed:16269684};
DE   AltName: Full=4,4'-diapophytoene synthase;
DE            Short=DAP synthase;
DE   AltName: Full=Diapophytoene synthase;
GN   Name=crtM; OrderedLocusNames=SAOUHSC_02879;
OS   Staphylococcus aureus (strain NCTC 8325).
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;
RA   Gillaspy A.F., Worrell V., Orvis J., Roe B.A., Dyer D.W.,
RA   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,
RL   Washington D.C. (2006).
RN   [2]
RP   CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND MUTAGENESIS OF 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
RT   C(40) 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 KINETIC PARAMETERS.
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).
CC   -!- FUNCTION: Catalyzes the head-to-head condensation of two molecules
CC       of farnesyl diphosphate (FPP) into the colorless C(30) carotenoid
CC       dehydrosqualene (4,4'-diapophytoene). This is the initial step in
CC       the biosynthesis of staphyloxanthin, an orange carotenoid present
CC       in most staphylococci strains. {ECO:0000269|PubMed:16269684}.
CC   -!- CATALYTIC ACTIVITY: 2 (2E,6E)-farnesyl diphosphate = 15-cis-4,4'-
CC       diapophytoene + 2 diphosphate. {ECO:0000269|PubMed:12426357,
CC       ECO:0000269|PubMed:16269684}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.2 uM for farnesyl diphosphate
CC         {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   -!- MISCELLANEOUS: CrtM is not functional in a C(40) pathway, however
CC       independent mutations on Phe-26, Trp-38 or Glu-180 are sufficient
CC       to 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-
CC       26 and Trp-38 appears to be harmful for the general performance of
CC       the enzyme.
CC   -!- MISCELLANEOUS: Upon coexpression with Erwinia
CC       geranylgeranyldiphosphate (GGDP) synthase, CrtM produces novel
CC       carotenoids with the asymmetrical C(35) backbone, such as 4-
CC       apophytoene, and the production of the natural product 4,4'-
CC       diapophytoene drops dramatically.
CC   -!- SIMILARITY: Belongs to the phytoene/squalene synthase family. CrtM
CC       subfamily. {ECO:0000305}.
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DR   EMBL; CP000253; ABD31876.1; -; Genomic_DNA.
DR   RefSeq; WP_000178307.1; NC_007795.1.
DR   RefSeq; YP_501333.1; NC_007795.1.
DR   ProteinModelPortal; Q2FV59; -.
DR   SMR; Q2FV59; -.
DR   STRING; 93061.SAOUHSC_02879; -.
DR   EnsemblBacteria; ABD31876; ABD31876; SAOUHSC_02879.
DR   GeneID; 3921549; -.
DR   KEGG; sao:SAOUHSC_02879; -.
DR   PATRIC; fig|93061.5.peg.2602; -.
DR   eggNOG; ENOG4105FE1; Bacteria.
DR   eggNOG; COG1562; LUCA.
DR   HOGENOM; HOG000220848; -.
DR   KO; K10208; -.
DR   OMA; MIAGQRM; -.
DR   BioCyc; SAUR93061:G1G5Y-2710-MONOMER; -.
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:0051996; F:squalene synthase activity; IEA:InterPro.
DR   GO; GO:0016117; P:carotenoid biosynthetic process; IEA:UniProtKB-KW.
DR   GO; GO:0006696; P:ergosterol biosynthetic process; IEA:InterPro.
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   Pfam; PF00494; SQS_PSY; 1.
DR   SUPFAM; SSF48576; SSF48576; 1.
DR   PROSITE; PS01044; SQUALEN_PHYTOEN_SYN_1; 1.
PE   1: Evidence at protein level;
KW   Carotenoid biosynthesis; Complete proteome; Reference proteome;
KW   Transferase.
FT   CHAIN         1    287       Dehydrosqualene synthase.
FT                                /FTId=PRO_0000282616.
FT   REGION       18     22       Farnesyl diphosphate 1 binding.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   REGION       45     48       Farnesyl diphosphate 2 binding.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   BINDING      41     41       Farnesyl diphosphate 1.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   BINDING     168    168       Farnesyl diphosphate 2.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   BINDING     171    171       Farnesyl diphosphate 1.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   BINDING     172    172       Farnesyl diphosphate 2.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   BINDING     248    248       Farnesyl diphosphate 1.
FT                                {ECO:0000250|UniProtKB:A9JQL9}.
FT   MUTAGEN      26     26       F->A,G,L,S: Decrease in C(30) carotene
FT                                synthase activity. C(40) carotene
FT                                synthase activity acquired.
FT                                {ECO:0000269|PubMed:12426357,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN      26     26       F->A: Decrease in C(30) and C(40)
FT                                carotene synthase activities; when
FT                                associated with A-38 or G-38.
FT                                {ECO:0000269|PubMed:12426357,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN      26     26       F->G: Decrease in C(30) and C(40)
FT                                carotene synthase activities; when
FT                                associated with A-38 or G-38.
FT                                {ECO:0000269|PubMed:12426357,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN      38     38       W->A: Decrease in C(30) and C(40)
FT                                carotene synthase activities; when
FT                                associated with A-26 or G-26.
FT                                {ECO:0000269|PubMed:12907743,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN      38     38       W->C: Decrease in C(30) carotene synthase
FT                                activity. C(40) carotene synthase
FT                                activity acquired.
FT                                {ECO:0000269|PubMed:12907743,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN      38     38       W->G: Decrease in C(30) and C(40)
FT                                carotene synthase activities; when
FT                                associated with A-26 or G-26.
FT                                {ECO:0000269|PubMed:12907743,
FT                                ECO:0000269|PubMed:14973014}.
FT   MUTAGEN     180    180       E->G: Slight increase in C(30) carotene
FT                                synthase activity. C(40) carotene
FT                                synthase activity acquired.
FT                                {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
//