title: Characterization of an isoflavonoid-specific prenyltransferase from Lupinus albus.
abstract: Prenylated flavonoids and isoflavonoids possess antimicrobial activity against fungal pathogens of plants. However, only a few plant flavonoid and isoflavonoid prenyltransferase genes have been identified to date. In this study, an isoflavonoid prenyltransferase gene, designated as LaPT1, was identified from white lupin (Lupinus albus). The deduced protein sequence of LaPT1 shared high homologies with known flavonoid and isoflavonoid prenyltransferases. The LaPT1 gene was mainly expressed in roots, a major site for constitutive accumulation of prenylated isoflavones in white lupin. LaPT1 is predicted to be a membrane-bound protein with nine transmembrane regions and conserved functional domains similar to other flavonoid and isoflavonoid prenyltransferases; it has a predicted chloroplast transit peptide and is plastid localized. A microsomal fraction containing recombinant LaPT1 prenylated the isoflavone genistein at the B-ring 3' position to produce isowighteone. The enzyme is also active with 2'-hydroxygenistein but has no activity with other flavonoid substrates. The apparent K(m) of recombinant LaPT1 for the dimethylallyl diphosphate prenyl donor is in a similar range to that of other flavonoid prenyltransferases, but the apparent catalytic efficiency with genistein is considerably higher. Removal of the transit peptide increased the apparent overall activity but also increased the K(m). Medicago truncatula hairy roots expressing LaPT1 accumulated isowighteone, a compound that is not naturally produced in this species, indicating a strategy for metabolic engineering of novel antimicrobial compounds in legumes.
authors.affiliation: Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, Oklahoma 73401, USA.
authors.forename: Guoan
authors.initials: G
authors.lastname: Shen
authors.forename: David
authors.initials: D
authors.lastname: Huhman
authors.forename: Zhentian
authors.initials: Z
authors.lastname: Lei
authors.forename: John
authors.initials: J
authors.lastname: Snyder
authors.forename: Lloyd W
authors.initials: LW
authors.lastname: Sumner
authors.forename: Richard A
authors.initials: RA
authors.lastname: Dixon
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D020869:Gene Expression Profiling; D017343:Genes, Plant; D019833:Genistein / Q000302:isolation & purification / Q000378:metabolism; D031297:Lupinus / Q000201:enzymology* / Q000235:genetics; D046913:Medicago truncatula / Q000201:enzymology / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D018087:Plastids / Q000201:enzymology / Q000235:genetics; D054876:Prenylation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D008565:Membrane Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins; D019833:Genistein; D004122:Dimethylallyltranstransferase