C93C2_GLYEC
ID C93C2_GLYEC Reviewed; 523 AA.
AC Q9SXS3;
DT 31-OCT-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 84.
DE RecName: Full=2-hydroxyisoflavanone synthase;
DE Short=2HI synthase;
DE EC=1.14.14.87 {ECO:0000269|PubMed:10557230, ECO:0000269|PubMed:12207646, ECO:0000269|PubMed:15809082};
DE AltName: Full=CYP Ge-8;
DE AltName: Full=Cytochrome P450 93C2;
DE AltName: Full=Isoflavonoid synthase;
GN Name=CYP93C2; Synonyms=IFS;
OS Glycyrrhiza echinata (Licorice).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Galegeae; Glycyrrhiza.
OX NCBI_TaxID=46348;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, INDUCTION BY
RP ELICITORS, AND SUBCELLULAR LOCATION.
RX PubMed=10557230; DOI=10.1104/pp.121.3.821;
RA Akashi T., Aoki T., Ayabe S.;
RT "Cloning and functional expression of a cytochrome P450 cDNA encoding 2-
RT hydroxyisoflavanone synthase involved in biosynthesis of the isoflavonoid
RT skeleton in licorice.";
RL Plant Physiol. 121:821-828(1999).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF ARG-104; GLN-106; VAL-119;
RP SER-305; SER-310 AND LYS-375, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP 3D-STRUCTURE MODELING.
RX PubMed=12207646; DOI=10.1046/j.1365-313x.2002.01378.x;
RA Sawada Y., Kinoshita K., Akashi T., Aoki T., Ayabe S.;
RT "Key amino acid residues required for aryl migration catalysed by the
RT cytochrome P450 2-hydroxyisoflavanone synthase.";
RL Plant J. 31:555-564(2002).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF SER-310; LEU-371 AND
RP LYS-375.
RX PubMed=15809082; DOI=10.1016/j.bbrc.2005.03.053;
RA Sawada Y., Ayabe S.;
RT "Multiple mutagenesis of P450 isoflavonoid synthase reveals a key active-
RT site residue.";
RL Biochem. Biophys. Res. Commun. 330:907-913(2005).
CC -!- FUNCTION: 2-hydroxyisoflavanone synthase, which catalyzes the
CC hydroxylation associated with 1,2-aryl migration of flavanones.
CC Converts liquiritigenin and naringenin into highly unstable precursors
CC of the isoflavones daidzein and genistein. Acts only on substrates with
CC (2S)-chirality. {ECO:0000269|PubMed:10557230,
CC ECO:0000269|PubMed:12207646, ECO:0000269|PubMed:15809082}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=liquiritigenin + O2 + reduced [NADPH--hemoprotein reductase] =
CC (2R,3S)-2,4',7-trihydroxyisoflavanone + H(+) + H2O + oxidized
CC [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:31723, Rhea:RHEA-
CC COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:28777,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:63325;
CC EC=1.14.14.87; Evidence={ECO:0000269|PubMed:10557230,
CC ECO:0000269|PubMed:12207646, ECO:0000269|PubMed:15809082};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2S)-naringenin + O2 + reduced [NADPH--hemoprotein reductase]
CC = 2-hydroxy-2,3-dihydrogenistein + H(+) + H2O + oxidized [NADPH--
CC hemoprotein reductase]; Xref=Rhea:RHEA:35487, Rhea:RHEA-COMP:11964,
CC Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17846, ChEBI:CHEBI:31080,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210; EC=1.14.14.87;
CC Evidence={ECO:0000269|PubMed:10557230, ECO:0000269|PubMed:12207646,
CC ECO:0000269|PubMed:15809082};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8 uM for liquiritigenin {ECO:0000269|PubMed:12207646};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:12207646};
CC -!- SUBCELLULAR LOCATION: Microsome membrane {ECO:0000305}; Single-pass
CC membrane protein {ECO:0000305}.
CC -!- INDUCTION: Up-regulated upon elicitation.
CC {ECO:0000269|PubMed:10557230}.
CC -!- MISCELLANEOUS: Lys-375 is essential for the intramolecular aryl
CC migration of a flavanone molecule to produce the isoflavonoid skeleton
CC (PubMed:12207646). Arg-104 and Leu-371 are essential for the correct
CC P450 conformation (PubMed:12207646, PubMed:15809082).
CC {ECO:0000305|PubMed:12207646, ECO:0000305|PubMed:15809082}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; AB023636; BAA76380.1; -; mRNA.
DR AlphaFoldDB; Q9SXS3; -.
DR SMR; Q9SXS3; -.
DR KEGG; ag:BAA76380; -.
DR GO; GO:0005783; C:endoplasmic reticulum; IEA:UniProtKB-KW.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0043231; C:intracellular membrane-bounded organelle; IDA:UniProtKB.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0102668; F:liquiritigenin,NADPH:oxygen oxidoreductase activity; IEA:UniProtKB-EC.
DR GO; GO:0102604; F:naringenin,NADPH:oxygen oxidoreductase activity; IEA:UniProtKB-EC.
DR GO; GO:0016709; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR GO; GO:0009717; P:isoflavonoid biosynthetic process; IDA:UniProtKB.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Endoplasmic reticulum; Heme; Iron; Membrane; Metal-binding; Microsome;
KW Monooxygenase; NADP; Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..523
FT /note="2-hydroxyisoflavanone synthase"
FT /id="PRO_0000419822"
FT TRANSMEM 2..22
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 450
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250"
FT MUTAGEN 104
FT /note="R->A: Total loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:12207646"
FT MUTAGEN 106
FT /note="Q->E: No effect on catalytic activity."
FT /evidence="ECO:0000269|PubMed:12207646"
FT MUTAGEN 119
FT /note="V->L: No effect on catalytic activity."
FT /evidence="ECO:0000269|PubMed:12207646"
FT MUTAGEN 305
FT /note="S->T: No effect on catalytic activity."
FT /evidence="ECO:0000269|PubMed:12207646"
FT MUTAGEN 310
FT /note="S->A,V: Decreased intramolecular 1,2-aryl migration
FT and increased production of 3-hydroxyflavanone and
FT flavone."
FT /evidence="ECO:0000269|PubMed:12207646,
FT ECO:0000269|PubMed:15809082"
FT MUTAGEN 310
FT /note="S->T: Decreased intramolecular 1,2-aryl migration
FT and increased production of 3-hydroxyflavanone and flavone.
FT 5% activity and production of mainly 3-hydroxyflavanone;
FT when associated with T-375. 0.1% activity and production of
FT only flavone; when associated with V-371 and T-375."
FT /evidence="ECO:0000269|PubMed:12207646,
FT ECO:0000269|PubMed:15809082"
FT MUTAGEN 371
FT /note="L->I: Production of mainly 3-hydroxyflavanone; when
FT associated with T-375."
FT /evidence="ECO:0000269|PubMed:15809082"
FT MUTAGEN 371
FT /note="L->V: Total loss of catalytic activity. 5% activity
FT and production of mainly 3-hydroxyflavanone; when
FT associated with T-375. 0.1% activity and production of only
FT flavone; when associated with T-310 and T-375."
FT /evidence="ECO:0000269|PubMed:15809082"
FT MUTAGEN 375
FT /note="K->A,R: Loss of intramolecular 1,2-aryl migration
FT and production of only 3-hydroxyflavanone."
FT /evidence="ECO:0000269|PubMed:12207646,
FT ECO:0000269|PubMed:15809082"
FT MUTAGEN 375
FT /note="K->T: Loss of intramolecular 1,2-aryl migration and
FT production of only 3-hydroxyflavanone. 5% activity and
FT production of mainly 3-hydroxyflavanone; when associated
FT with T-310; I-371 or V-371. 0.1% activity and production of
FT only flavone; when associated with T-310 and V-371."
FT /evidence="ECO:0000269|PubMed:12207646,
FT ECO:0000269|PubMed:15809082"
SQ SEQUENCE 523 AA; 59429 MW; A0E7F1127847BAB2 CRC64;
MLVELAITLL VIALFIHLRP TLSAKSKSLR HLPNPPSPKP RLPFVGHLHL LDKPLLHYSL
IDLSKRYGPL YSLYFGSMPT VVASTPELFK LFLQTHEASS FNTRFQTSAI RRLTYDNSVA
MVPFGPYWKF IRKLIMNDLL NATTVNKLRP LRSQEIRKVL RVMAQSAESQ VPLNVTEELL
KWTNSTISRM MLGEAEEIRD IARDVLKIFG EYSLTDFIWP LKKLKVGQYE KRIDDIFNRF
DPVIERVIKK RQEIRKKRKE RNGEIEEGEQ SVVFLDTLLD FAEDETMEIK ITKEQIKGLV
VDFFSAGTDS TAVATDWALS ELINNPRVFQ KAREEIDAVV GKDRLVDEAD VQNLPYIRSI
VKETFRMHPP LPVVKRKCVQ ECEVDGYVIP EGALILFNVW AVGRDPKYWD RPTEFRPERF
LENVGEGDQA VDLRGQHFQL LPFGSGRRMC PGVNLATAGM ATLLASVIQC FDLSVVGPQG
KILKGNDAKV SMEERAGLTV PRAHNLICVP VARSSAVPKL FSS
