ID DFRA_MALDO Reviewed; 348 AA. AC Q9XES5; Q84KP1; Q8L5N3; DT 24-MAR-2009, integrated into UniProtKB/Swiss-Prot. DT 01-NOV-1999, sequence version 1. DT 24-JAN-2024, entry version 80. DE RecName: Full=Bifunctional dihydroflavonol 4-reductase/flavanone 4-reductase; DE AltName: Full=Dihydroflavonol 4-reductase; DE Short=DFR; DE EC=1.1.1.219 {ECO:0000269|PubMed:12667486}; DE AltName: Full=Dihydrokaempferol 4-reductase; DE AltName: Full=Flavanone 4-reductase; DE Short=FNR; DE EC=1.1.1.234 {ECO:0000269|PubMed:12667486}; GN Name=DFR; OS Malus domestica (Apple) (Pyrus malus). OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae; OC rosids; fabids; Rosales; Rosaceae; Amygdaloideae; Maleae; Malus. OX NCBI_TaxID=3750; RN [1] RP NUCLEOTIDE SEQUENCE [MRNA]. RC STRAIN=cv. Fuji; TISSUE=Peelings; RA Lee J.-R., Hong S.-T., Yoo Y.G., Kim S.-R.; RT "Molecular cloning and expression of anthocyanin biosynthesis genes from RT 'Fuji apple'."; RL Submitted (DEC-1998) to the EMBL/GenBank/DDBJ databases. RN [2] RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, AND RP BIOPHYSICOCHEMICAL PROPERTIES. RC STRAIN=cv. M9, and cv. Weirouge; RX PubMed=12667486; DOI=10.1016/s0003-9861(03)00013-4; RA Fischer T.C., Halbwirth H., Meisel B., Stich K., Forkmann G.; RT "Molecular cloning, substrate specificity of the functionally expressed RT dihydroflavonol 4-reductases from Malus domestica and Pyrus communis RT cultivars and the consequences for flavonoid metabolism."; RL Arch. Biochem. Biophys. 412:223-230(2003). RN [3] RP NUCLEOTIDE SEQUENCE [MRNA] OF 4-317, AND DEVELOPMENTAL STAGE. RC STRAIN=cv. Jonathan; TISSUE=Peelings; RA Honda C., Kotoda N., Wada M., Kondo S., Kobayashi S., Soejima J., Zhang Z., RA Tsuda T., Moriguchi T.; RT "Anthocyanin biosynthetic genes are coordinately expressed during red RT coloration in apple skin."; RL Plant Physiol. Biochem. 40:955-962(2002). CC -!- FUNCTION: Bifunctional enzyme involved in flavonoid metabolism. May use CC dihydroquercetin, dihydrokaempferol, eriodictyol, garbanzol (5- CC deoxydihydrokaempferol), dihydrofisetin (5-deoxydihydroquercetin), CC naringenin to a low extent (10%), but not 5-deoxynaringenin or butin CC (5-deoxyeriodictyol) as substrate. {ECO:0000269|PubMed:12667486}. CC -!- CATALYTIC ACTIVITY: CC Reaction=a (2R,3S,4S)-leucoanthocyanidin + NADP(+) = a (2R,3R)- CC dihydroflavonol + H(+) + NADPH; Xref=Rhea:RHEA:54444, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, CC ChEBI:CHEBI:138176, ChEBI:CHEBI:138188; EC=1.1.1.219; CC Evidence={ECO:0000269|PubMed:12667486}; CC -!- CATALYTIC ACTIVITY: CC Reaction=(2S)-flavan-4-ol + NADP(+) = (2S)-flavanone + H(+) + NADPH; CC Xref=Rhea:RHEA:11228, ChEBI:CHEBI:15378, ChEBI:CHEBI:15605, CC ChEBI:CHEBI:15606, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; CC EC=1.1.1.234; Evidence={ECO:0000269|PubMed:12667486}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=3.9 uM for eriodictyol {ECO:0000269|PubMed:12667486}; CC KM=34 uM for dihydrokaempferol {ECO:0000269|PubMed:12667486}; CC KM=26 uM for dihydroquercetin {ECO:0000269|PubMed:12667486}; CC Vmax=4.6 nmol/sec/g enzyme toward eriodictyol CC {ECO:0000269|PubMed:12667486}; CC Vmax=186 nmol/sec/g enzyme toward dihydrokaempferol CC {ECO:0000269|PubMed:12667486}; CC Vmax=91 nmol/sec/g enzyme toward dihydroquercetin CC {ECO:0000269|PubMed:12667486}; CC pH dependence: CC Optimum pH is 5.75 with dihydroquercetin as substrate. CC {ECO:0000269|PubMed:12667486}; CC -!- DEVELOPMENTAL STAGE: Expressed during fruit ripening. CC {ECO:0000269|Ref.3}. CC -!- SIMILARITY: Belongs to the NAD(P)-dependent epimerase/dehydratase CC family. Dihydroflavonol-4-reductase subfamily. {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AF117268; AAD26204.1; -; mRNA. DR EMBL; AY227729; AAO39817.1; -; mRNA. DR EMBL; AY227728; AAO39816.1; -; mRNA. DR EMBL; AB074488; BAB92999.1; -; mRNA. DR RefSeq; NP_001280868.1; NM_001293939.1. DR AlphaFoldDB; Q9XES5; -. DR SMR; Q9XES5; -. DR EnsemblPlants; mRNA:MD15G0020300; mRNA:MD15G0020300; MD15G0020300. DR GeneID; 103450464; -. DR Gramene; mRNA:MD15G0020300; mRNA:MD15G0020300; MD15G0020300. DR KEGG; mdm:103450464; -. DR OrthoDB; 1694166at2759; -. DR BRENDA; 1.1.1.234; 3164. DR GO; GO:0045552; F:dihydrokaempferol 4-reductase activity; IEA:UniProtKB-EC. DR GO; GO:0047890; F:flavanone 4-reductase activity; IEA:UniProtKB-EC. DR GO; GO:0009813; P:flavonoid biosynthetic process; IEA:UniProtKB-KW. DR CDD; cd08958; FR_SDR_e; 1. DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1. DR InterPro; IPR001509; Epimerase_deHydtase. DR InterPro; IPR036291; NAD(P)-bd_dom_sf. DR PANTHER; PTHR10366:SF613; DIHYDROFLAVONOL 4-REDUCTASE; 1. DR PANTHER; PTHR10366; NAD DEPENDENT EPIMERASE/DEHYDRATASE; 1. DR Pfam; PF01370; Epimerase; 1. DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1. PE 1: Evidence at protein level; KW Flavonoid biosynthesis; NADP; Oxidoreductase. FT CHAIN 1..348 FT /note="Bifunctional dihydroflavonol 4-reductase/flavanone FT 4-reductase" FT /id="PRO_0000367056" FT BINDING 44 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT /evidence="ECO:0000250|UniProtKB:A0A059TC02" FT BINDING 163 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT /evidence="ECO:0000250|UniProtKB:A0A059TC02" FT VARIANT 347 FT /note="V -> A (in strain: cv. M9)" SQ SEQUENCE 348 AA; 39030 MW; 02056428FB2F9372 CRC64; MGSESESVCV TGASGFIGSW LVMRLLEHGY TVRATVRDPT NQKKVKHLLD LPKAETHLTL WKADLADEGS FDEAIQGCSG VFHVATPMDF ESKDPENEVI KPTINGLLDI LKACQKAKTV RKLVFTSSAG TVNVEEHQKP VYDESNWSDV EFCRSVKMTG WMYFVSKTLA EQAAWKYAKE NNIDFITIIP TLVIGPFLMP SMPPSLITGL SPILRNESHY GIIKQGQYVH LDDLCLSHIY LYEHPKAEGR YICSSHDATI HELVKMLREK YPEYNIPTKF KGIDDNLEPV HFSSKKLREI GFEFKYSLED MFVGAVDACR AKGLIPIPIP AEKTEAAEES NLVDVKVG //