CRTM_STAA8
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 03-AUG-2022, entry version 95.
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}.
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DR EMBL; CP000253; ABD31876.1; -; Genomic_DNA.
DR RefSeq; WP_000178307.1; NZ_LS483365.1.
DR RefSeq; YP_501333.1; NC_007795.1.
DR AlphaFoldDB; Q2FV59; -.
DR SMR; Q2FV59; -.
DR STRING; 1280.SAXN108_2813; -.
DR EnsemblBacteria; ABD31876; ABD31876; SAOUHSC_02879.
DR GeneID; 3921549; -.
DR KEGG; sao:SAOUHSC_02879; -.
DR PATRIC; fig|93061.5.peg.2602; -.
DR eggNOG; COG1562; Bacteria.
DR HOGENOM; CLU_037269_1_3_9; -.
DR OMA; KLYCYRV; -.
DR SABIO-RK; Q2FV59; -.
DR UniPathway; UPA00029; UER00556.
DR PRO; PR:Q2FV59; -.
DR Proteomes; UP000008816; Chromosome.
DR GO; GO:0004311; F:farnesyltranstransferase 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:0016117; P:carotenoid biosynthetic process; 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; IPR044843; Trans_IPPS_bact-type.
DR InterPro; IPR033904; Trans_IPPS_HH.
DR Pfam; PF00494; SQS_PSY; 1.
DR SFLD; SFLDG01212; Phytoene_synthase_like; 1.
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 BINDING 18..21
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 41
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 45
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 45
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 48
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 52
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 165
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 168
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 171
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 172
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:A9JQL9"
FT BINDING 248
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /ligand_label="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
