CBDAS_CANSA
ID CBDAS_CANSA Reviewed; 544 AA.
AC A6P6V9;
DT 06-FEB-2013, integrated into UniProtKB/Swiss-Prot.
DT 21-AUG-2007, sequence version 1.
DT 03-AUG-2022, entry version 56.
DE RecName: Full=Cannabidiolic acid synthase {ECO:0000303|PubMed:8663284};
DE Short=CBDA synthase {ECO:0000303|PubMed:8663284};
DE EC=1.21.3.8 {ECO:0000269|PubMed:17544411, ECO:0000269|PubMed:8663284};
DE Flags: Precursor;
GN Name=CBDAS {ECO:0000303|PubMed:8663284};
OS Cannabis sativa (Hemp) (Marijuana).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Rosales; Cannabaceae; Cannabis.
OX NCBI_TaxID=3483;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 29-53; 395-408 AND 428-442,
RP FUNCTION, CATALYTIC ACTIVITY, GLYCOSYLATION, AND MUTAGENESIS OF HIS-114.
RX PubMed=17544411; DOI=10.1016/j.febslet.2007.05.043;
RA Taura F., Sirikantaramas S., Shoyama Y., Yoshikai K., Shoyama Y.,
RA Morimoto S.;
RT "Cannabidiolic-acid synthase, the chemotype-determining enzyme in the
RT fiber-type Cannabis sativa.";
RL FEBS Lett. 581:2929-2934(2007).
RN [2]
RP PROTEIN SEQUENCE OF 29-44, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, SUBUNIT, SUBSTRATE SPECIFICITY, TISSUE
RP SPECIFICITY, AND SUBCELLULAR LOCATION.
RX PubMed=8663284; DOI=10.1074/jbc.271.29.17411;
RA Taura F., Morimoto S., Shoyama Y.;
RT "Purification and characterization of cannabidiolic-acid synthase from
RT Cannabis sativa L. Biochemical analysis of a novel enzyme that catalyzes
RT the oxidocyclization of cannabigerolic acid to cannabidiolic acid.";
RL J. Biol. Chem. 271:17411-17416(1996).
RN [3]
RP REVIEW.
RX PubMed=17712812; DOI=10.1002/cbdv.200790145;
RA Taura F., Sirikantaramas S., Shoyama Y., Shoyama Y., Morimoto S.;
RT "Phytocannabinoids in Cannabis sativa: recent studies on biosynthetic
RT enzymes.";
RL Chem. Biodivers. 4:1649-1663(2007).
RN [4]
RP PATHWAY, AND REVIEW.
RX PubMed=30468448; DOI=10.1039/c8np00077h;
RA Liu Y., Jing S.-X., Luo S.-H., Li S.-H.;
RT "Non-volatile natural products in plant glandular trichomes: chemistry,
RT biological activities and biosynthesis.";
RL Nat. Prod. Rep. 36:626-665(2019).
CC -!- FUNCTION: Oxidoreductase involved in the biosynthesis of cannabinoids-
CC related terpenophenolic natural products, which have pharmacological
CC activity (PubMed:17544411, PubMed:8663284). Catalyzes the
CC stereoselective oxidative cyclization of the monoterpene moiety in
CC cannabigerolic acid (CBGA), producing cannabidiolate (CBDA), the major
CC cannabioid in fiber-type Cannabis plants (PubMed:17544411,
CC PubMed:8663284). Can also use cannabinerolic acid as substrate, but not
CC cannabigerol or cannabinerol (PubMed:17544411, PubMed:8663284).
CC {ECO:0000269|PubMed:17544411, ECO:0000269|PubMed:8663284}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cannabigerolate + O2 = cannabidiolate + H2O2;
CC Xref=Rhea:RHEA:34411, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:66962, ChEBI:CHEBI:67136; EC=1.21.3.8;
CC Evidence={ECO:0000269|PubMed:17544411, ECO:0000269|PubMed:8663284};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:34412;
CC Evidence={ECO:0000269|PubMed:17544411, ECO:0000269|PubMed:8663284};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000250|UniProtKB:Q8GTB6};
CC Note=Binds 1 FAD per subunit in a bicovalent manner.
CC {ECO:0000250|UniProtKB:Q8GTB6};
CC -!- ACTIVITY REGULATION: Inhibited by Hg(2+). {ECO:0000269|PubMed:8663284}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.137 mM for cannabigerolic acid {ECO:0000269|PubMed:8663284};
CC KM=0.206 mM for cannabinerolic acid {ECO:0000269|PubMed:8663284};
CC Vmax=2.57 nmol/sec/mg enzyme with cannabigerolic acid as substrate
CC {ECO:0000269|PubMed:8663284};
CC Vmax=0.39 nmol/sec/mg enzyme with cannabinerolic acid as substrate
CC {ECO:0000269|PubMed:8663284};
CC pH dependence:
CC Optimum pH is 5.0. {ECO:0000269|PubMed:8663284};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000303|PubMed:30468448}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:8663284}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:8663284}. Secreted,
CC extracellular space, apoplast {ECO:0000250}. Note=Probably sorted from
CC the secretory cells into the storage cavity of glandular trichomes.
CC -!- TISSUE SPECIFICITY: Expressed in young leaves.
CC {ECO:0000269|PubMed:8663284}.
CC -!- PTM: Glycosylated. {ECO:0000269|PubMed:17544411}.
CC -!- PTM: The FAD cofactor is bound via a bicovalent 6-S-cysteinyl, 8alpha-
CC N1-histidyl FAD linkage. {ECO:0000250|UniProtKB:Q8GTB6}.
CC -!- MISCELLANEOUS: CBDA synthase might contribute to the self-defense of
CC Cannabis plants by producing both CBDA and hydrogen peroxide, but since
CC these reaction products are toxic to the plant itself, CBDA synthase is
CC probably secreted from secretory cells into the storage cavity to avoid
CC cellular damage. {ECO:0000250}.
CC -!- SIMILARITY: Belongs to the oxygen-dependent FAD-linked oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; AB292682; BAF65033.1; -; mRNA.
DR AlphaFoldDB; A6P6V9; -.
DR SMR; A6P6V9; -.
DR KEGG; ag:BAF65033; -.
DR BioCyc; MetaCyc:MON-12034; -.
DR BRENDA; 1.21.3.8; 1159.
DR UniPathway; UPA00213; -.
DR GO; GO:0048046; C:apoplast; IEA:UniProtKB-SubCell.
DR GO; GO:0102779; F:cannabidiolate synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:1901696; P:cannabinoid biosynthetic process; TAS:UniProtKB.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.30.43.10; -; 1.
DR Gene3D; 3.30.465.10; -; 1.
DR InterPro; IPR012951; BBE.
DR InterPro; IPR016166; FAD-bd_PCMH.
DR InterPro; IPR036318; FAD-bd_PCMH-like_sf.
DR InterPro; IPR016167; FAD-bd_PCMH_sub1.
DR InterPro; IPR016169; FAD-bd_PCMH_sub2.
DR InterPro; IPR006094; Oxid_FAD_bind_N.
DR Pfam; PF08031; BBE; 1.
DR Pfam; PF01565; FAD_binding_4; 1.
DR SUPFAM; SSF56176; SSF56176; 1.
DR PROSITE; PS51387; FAD_PCMH; 1.
PE 1: Evidence at protein level;
KW Apoplast; Direct protein sequencing; Disulfide bond; FAD; Flavoprotein;
KW Glycoprotein; Oxidoreductase; Secreted; Signal.
FT SIGNAL 1..28
FT /evidence="ECO:0000269|PubMed:17544411,
FT ECO:0000269|PubMed:8663284"
FT CHAIN 29..544
FT /note="Cannabidiolic acid synthase"
FT /id="PRO_0000421144"
FT DOMAIN 77..251
FT /note="FAD-binding PCMH-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00718"
FT ACT_SITE 483
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 109..115
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 120
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 176
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 180..184
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 190
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 236
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 241
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 291
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 416
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 441
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:G2QG48"
FT BINDING 480..482
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT CARBOHYD 45
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 65
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 168
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 296
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 304
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 328
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 498
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT DISULFID 37..99
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT CROSSLNK 114..176
FT /note="6-(S-cysteinyl)-8alpha-(pros-histidyl)-FAD (His-
FT Cys)"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT MUTAGEN 114
FT /note="H->A: Loss of FAD binding and loss of catalytic
FT activity."
FT /evidence="ECO:0000269|PubMed:17544411"
SQ SEQUENCE 544 AA; 62237 MW; 199378287274F156 CRC64;
MKCSTFSFWF VCKIIFFFFS FNIQTSIANP RENFLKCFSQ YIPNNATNLK LVYTQNNPLY
MSVLNSTIHN LRFTSDTTPK PLVIVTPSHV SHIQGTILCS KKVGLQIRTR SGGHDSEGMS
YISQVPFVIV DLRNMRSIKI DVHSQTAWVE AGATLGEVYY WVNEKNENLS LAAGYCPTVC
AGGHFGGGGY GPLMRNYGLA ADNIIDAHLV NVHGKVLDRK SMGEDLFWAL RGGGAESFGI
IVAWKIRLVA VPKSTMFSVK KIMEIHELVK LVNKWQNIAY KYDKDLLLMT HFITRNITDN
QGKNKTAIHT YFSSVFLGGV DSLVDLMNKS FPELGIKKTD CRQLSWIDTI IFYSGVVNYD
TDNFNKEILL DRSAGQNGAF KIKLDYVKKP IPESVFVQIL EKLYEEDIGA GMYALYPYGG
IMDEISESAI PFPHRAGILY ELWYICSWEK QEDNEKHLNW IRNIYNFMTP YVSKNPRLAY
LNYRDLDIGI NDPKNPNNYT QARIWGEKYF GKNFDRLVKV KTLVDPNNFF RNEQSIPPLP
RHRH
