Identification and characterization of isoflavonoid specific glycosyltransferase and malonyltransferase from soybean seeds.
Isoflavonoids are a diverse group of biologically active natural products that accumulate in soybean seeds during development. The majority of isoflavonoids are accumulated in the form of their glyco- and malonyl-conjugates in soybean seeds. The conjugation step confers stability and solubility to isoflavone aglycones enabling their compartmentalization to vacuoles or transport to the site of accumulation. A functional genomic approach was used to identify isoflavonoid specific glycosyltransferase (UGT) and malonyltransferase (MT) from soybean (Glycine max) seeds. An expressed sequence tag database for soybean was searched by key words to make a list of candidate genes. The full-length cDNAs for candidate UGTs and MTs were obtained and cloned into an expression vector for the production of recombinant enzymes. The in vitro enzymatic activity assays were conducted for recombinant UGTs and MTs using uridine diphosphate glucose and malonyl CoA, respectively, as donors with isoflavone substrates. Among several recombinant enzymes, UGT73F2 showed glycosylation activity towards all three soybean isoflavone aglycones and GmMT7 exhibited malonylation activity towards isoflavone glycosides. The subcellular localization study revealed both UGT73F2 and GmMT7 to be in the cytoplasm. The transcripts and protein accumulation patterns for UGT73F2 and GmMT7 genes have provided further support for their in planta function.
Dhaubhadel
Sangeeta
S
Southern Crop Protection and Food Research Center, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, Ontario, N5V 4T3, Canada.
Farhangkhoee
Mana
M
Chapman
Ralph
R
D000595:Amino Acid Sequence; D018076:DNA, Complementary / Q000235:genetics; D005419:Flavonoids / Q000737:chemistry* / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D041743:Herb-Drug Interactions / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012639:Seeds / Q000201:enzymology*; D013025:Glycine max / Q000201:enzymology*; D013379:Substrate Specificity
D018076:DNA, Complementary; D005419:Flavonoids; D010940:Plant Proteins; D016695:Glycosyltransferases