Vicianin hydrolase is a novel cyanogenic beta-glycosidase specific to beta-vicianoside (6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside) in seeds of Vicia angustifolia.
The cyanogenic disaccharide glycoside, vicianin [mandelonitrile beta-vicianoside (6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside)], is accumulated in seeds of Vicia angustifolia var. segetalis. Vicianin hydrolase (VH) catalyzes the hydrolysis of vicianin into mandelonitrile and a disaccharide vicianose. VH was purified from the seeds using DEAE-, CM- and Con A-Sepharose chromatography, and the molecular mass of the purified VH was estimated to be 56 kDa on SDS-PAGE. The N-terminal amino acid sequence of the purified VH was determined, and a cDNA encoding VH was obtained. The deduced VH protein consists of a 509 amino acid polypeptide containing a putative secretion signal peptide. It shares about 50% identity with various kinds of plant beta-glycosidases including tea leaf beta-primeverosidase and furcatin hydrolase, and is classified in family 1 of the glycosyl hydrolases. The VH transcript was detected abundantly in seeds and moderately in flowers, but only slightly in leaves, stems and roots, indicating that the organ distribution of VH expression is similar to that of the substrate vicianin. The recombinant VH was produced in insect cells with a baculovirus system, and was compared with the native VH in terms of substrate specificity. Both enzymes hydrolyzed vicianin to release vicianose, demonstrating that VH is a disaccharide-specific beta-glycosidase. VH also hydrolyzed the mandelonitrile beta-glucoside prunasin to some extent but did not hydrolyze the gentiobioside amygdalin, both of which contain the same aglycone as vicianin. Thus, VH is a unique cyanogenic glycosidase showing high glycone specificity for the disaccharide vicianoside.
Ahn
Young Ock
YO
Institute for Chemical Research, Kyoto University, Uji, Kyoto, 611-0011 Japan.
Saino
Hiromichi
H
Mizutani
Masaharu
M
Shimizu
Bun-ichi
B
Sakata
Kanzo
K
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003486:Cyanides / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004187:Disaccharides / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D005959:Glucosidases / Q000235:genetics / Q000378:metabolism*; D007313:Insecta; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism*; D013379:Substrate Specificity; D031306:Vicia / Q000201:enzymology* / Q000235:genetics
C524320:6-O-arabinopyranosylglucopyranoside; D003486:Cyanides; D018076:DNA, Complementary; D018744:DNA, Plant; D004187:Disaccharides; D010940:Plant Proteins; D005959:Glucosidases