CBCAS_CANSA
ID CBCAS_CANSA Reviewed; 545 AA.
AC Q33DQ2; A0A3G5EA46; A0A3G5EAK0;
DT 06-FEB-2013, integrated into UniProtKB/Swiss-Prot.
DT 06-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 53.
DE RecName: Full=Cannabichromenic acid synthase {ECO:0000303|PubMed:9862135, ECO:0000303|Ref.3, ECO:0000303|Ref.5};
DE EC=1.21.99.- {ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3, ECO:0000269|Ref.5};
DE Flags: Precursor;
GN Name=CBCAS {ECO:0000303|PubMed:9862135, ECO:0000303|Ref.3,
GN ECO:0000303|Ref.5};
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 [GENOMIC DNA], FUNCTION, POLYMORPHISM, AND VARIANTS.
RC STRAIN=001, 010, 013, 020, 053, 054, and 069;
RX PubMed=16143478; DOI=10.1016/j.forsciint.2005.07.005;
RA Kojoma M., Seki H., Yoshida S., Muranaka T.;
RT "DNA polymorphisms in the tetrahydrocannabinolic acid (THCA) synthase gene
RT in 'drug-type' and 'fiber-type' Cannabis sativa L.";
RL Forensic Sci. Int. 159:132-140(2006).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND VARIANTS.
RC STRAIN=01, and 03;
RA El Alaoui M.A., Stambouli H., El Bouri A., Melloul M., Alaoui S.A.,
RA Soulaymani A., El Fahime E.;
RT "Study of moroccan Cannabis sativa DNA polymorphism in the
RT tetrahydrocannabinolic acid (thc) synthase gene.";
RL Submitted (JAN-2012) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, AND CATALYTIC ACTIVITY.
RA Page J.E., Stout J.M.;
RT "Cannabichromenic acid synthase from Cannabis sativa.";
RL Patent number EP3161140, 27-JUN-2014.
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=cv. Carmagnola4, cv. Ermes1, cv. Santhica2, and cv. Uso2;
RX PubMed=31718081; DOI=10.3390/plants8110496;
RA Cascini F., Farcomeni A., Migliorini D., Baldassarri L., Boschi I.,
RA Martello S., Amaducci S., Lucini L., Bernardi J.;
RT "Highly Predictive Genetic Markers Distinguish Drug-Type from Fiber-Type
RT Cannabis sativa L.";
RL Plants (Basel) 8:496-508(2019).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND TISSUE
RP SPECIFICITY.
RX DOI=10.1021/np970210y;
RA Morimoto S., Komatsu K., Taura F., Shoyama Y.;
RT "Enzymological evidence for cannabichromenic acid biosynthesis.";
RL J. Nat. Prod. 60:854-857(1997).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND
RP ACTIVITY REGULATION.
RX PubMed=9862135; DOI=10.1016/s0031-9422(98)00278-7;
RA Morimoto S., Komatsu K., Taura F., Shoyama Y.;
RT "Purification and characterization of cannabichromenic acid synthase from
RT Cannabis sativa.";
RL Phytochemistry 49:1525-1529(1998).
RN [7]
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 [8]
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 (Ref.5, Ref.3, PubMed:9862135). Catalyzes the oxidative
CC cyclization of the monoterpene moiety in cannabigerolic acid (CBGA)
CC producing cannabichromenic acid (CBCA), which exhibits anti-
CC inflammatory, antifungal and antimicrobial effects (Ref.5, Ref.3,
CC PubMed:9862135). Can also use cannabinerolic acid (CBNRA) as substrate
CC (PubMed:9862135). Unable to catalyze the production of delta(9)-
CC tetrahydrocannabinolate (THCA), the precursor of Delta(1)-
CC tetrahydrocannabinol (PubMed:16143478). {ECO:0000269|PubMed:16143478,
CC ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3, ECO:0000269|Ref.5}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=A + cannabigerolate = AH2 + cannabichromenate;
CC Xref=Rhea:RHEA:60432, ChEBI:CHEBI:13193, ChEBI:CHEBI:17499,
CC ChEBI:CHEBI:66962, ChEBI:CHEBI:167554;
CC Evidence={ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3,
CC ECO:0000269|Ref.5};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:60433;
CC Evidence={ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3,
CC ECO:0000269|Ref.5};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=A + cannabinerolate = AH2 + cannabichromenate;
CC Xref=Rhea:RHEA:66904, ChEBI:CHEBI:13193, ChEBI:CHEBI:17499,
CC ChEBI:CHEBI:66961, ChEBI:CHEBI:167554;
CC Evidence={ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3,
CC ECO:0000269|Ref.5};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66905;
CC Evidence={ECO:0000269|PubMed:9862135, ECO:0000269|Ref.3,
CC ECO:0000269|Ref.5};
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 hydrogen peroxide H(2)O(2) and FMN
CC (PubMed:9862135). Completely repressed activity by Hg(2+)
CC (PubMed:9862135). {ECO:0000269|PubMed:9862135}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=23 uM for cannabigerolic acid {ECO:0000269|PubMed:9862135};
CC KM=33 uM for cannabinerolic acid {ECO:0000269|PubMed:9862135};
CC Vmax=0.63 nmol/sec/mg enzyme with cannabigerolic acid as substrate
CC {ECO:0000269|PubMed:9862135};
CC Vmax=0.14 nmol/sec/mg enzyme with cannabinerolic acid as substrate
CC {ECO:0000269|PubMed:9862135};
CC Note=kcat is 0.04 sec(-1) with cannabigerolic acid as substrate.
CC {ECO:0000269|PubMed:9862135};
CC pH dependence:
CC Optimum pH is 6.5. {ECO:0000269|Ref.5};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000303|PubMed:30468448}.
CC -!- SUBUNIT: Monomer (By similarity). May form homodimer (PubMed:9862135).
CC {ECO:0000250|UniProtKB:Q8GTB6, ECO:0000269|PubMed:9862135}.
CC -!- TISSUE SPECIFICITY: Expressed in young leaves. {ECO:0000269|Ref.5}.
CC -!- PTM: The FAD cofactor is bound via a bicovalent 6-S-cysteinyl, 8alpha-
CC N1-histidyl FAD linkage. {ECO:0000250|UniProtKB:Q8GTB6}.
CC -!- POLYMORPHISM: Several isoforms of the inactive tetrahydrocannabinolic
CC acid synthase found in the 'fiber-type' cannabis plants exist due to
CC polymorphism. {ECO:0000269|PubMed:16143478}.
CC -!- SIMILARITY: Belongs to the oxygen-dependent FAD-linked oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; JQ437493; AFI24257.1; -; Genomic_DNA.
DR EMBL; JQ437494; AFI24258.1; -; Genomic_DNA.
DR EMBL; JQ437495; AFI24259.1; -; Genomic_DNA.
DR EMBL; JQ437496; AFI24260.1; -; Genomic_DNA.
DR EMBL; AB212830; BAE48242.1; -; Genomic_DNA.
DR EMBL; AB212831; BAE48243.1; -; Genomic_DNA.
DR EMBL; MG996402; AYW35088.1; -; Genomic_DNA.
DR EMBL; MG996405; AYW35091.1; -; Genomic_DNA.
DR EMBL; MG996409; AYW35095.1; -; Genomic_DNA.
DR EMBL; MG996411; AYW35097.1; -; Genomic_DNA.
DR AlphaFoldDB; Q33DQ2; -.
DR SMR; Q33DQ2; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
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 Disulfide bond; FAD; Flavoprotein; Glycoprotein; Oxidoreductase; Signal.
FT SIGNAL 1..28
FT /evidence="ECO:0000255"
FT CHAIN 29..545
FT /note="Cannabichromenic acid synthase"
FT /id="PRO_0000421143"
FT DOMAIN 77..251
FT /note="FAD-binding PCMH-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00718"
FT ACT_SITE 484
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 292
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 417
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT BINDING 442
FT /ligand="cannabigerolate"
FT /ligand_id="ChEBI:CHEBI:66962"
FT /evidence="ECO:0000250|UniProtKB:G2QG48"
FT BINDING 481..483
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q8GTB6"
FT CARBOHYD 65
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 89
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 297
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 305
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 329
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 467
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 499
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 CONFLICT 5
FT /note="A -> T (in Ref. 4; AYW35088/AYW35091/AYW35095/
FT AYW35097 and 3)"
FT /evidence="ECO:0000305"
FT CONFLICT 244
FT /note="W -> C (in Ref. 3)"
FT /evidence="ECO:0000305"
FT CONFLICT 543
FT /note="H -> R (in Ref. 3)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 545 AA; 61990 MW; 1A19BECE99931796 CRC64;
MNCSAFSFWF VCKIIFFFLS FNIQISIANP QENFLKCFSE YIPNNPANPK FIYTQHDQLY
MSVLNSTIQN LRFTSDTTPK PLVIVTPSNV SHIQASILCS KKVGLQIRTR SGGHDAEGLS
YISQVPFAIV DLRNMHTVKV DIHSQTAWVE AGATLGEVYY WINEMNENFS FPGGYCPTVG
VGGHFSGGGY GALMRNYGLA ADNIIDAHLV NVDGKVLDRK SMGEDLFWAI RGGGGENFGI
IAAWKIKLVV VPSKATIFSV KKNMEIHGLV KLFNKWQNIA YKYDKDLMLT THFRTRNITD
NHGKNKTTVH GYFSSIFLGG VDSLVDLMNK SFPELGIKKT DCKELSWIDT TIFYSGVVNY
NTANFKKEIL LDRSAGKKTA FSIKLDYVKK LIPETAMVKI LEKLYEEEVG VGMYVLYPYG
GIMDEISESA IPFPHRAGIM YELWYTATWE KQEDNEKHIN WVRSVYNFTT PYVSQNPRLA
YLNYRDLDLG KTNPESPNNY TQARIWGEKY FGKNFNRLVK VKTKADPNNF FRNEQSIPPL
PPHHH
