title: Purification and characterization of alpha-galactosidase from sunflower seeds.
abstract: From 100 g sunflower seeds, 1.2 mg purified alpha-galactosidase was obtained with an overall yield of 51%. The alpha-galactosidase acted on both terminal alpha-galactosyl residues and side-chain alpha-galactosyl residues of the galactomanno-oligosaccharides and galactomannans. The cDNA coding for sunflower alpha-galactosidase was cloned and the deduced amino acid sequence revealed that the mature enzyme consisted of 363 amino acid residues with a molecular weight of 40,263. Seven cysteine residues were found but no putative N-glycosylation sites were present in the sequence. The deduced amino acid sequences of mature enzyme and alpha-galactosidases from coffee, guar and Mortierella vinacea alpha-galactosidase II showed over 81%, 77%, and 47% homology, respectively. These enzymes are classified into the third group in which the enzyme has no insertion sequence and a broad specificity on galactomanno-oligosaccharides compared to the other groups.
authors.affiliation: Institute of Applied Biochemistry, University of Tsukuba, Ibaraki 305-8572, Japan.
authors.forename: Wook-Dong
authors.initials: WD
authors.lastname: Kim
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kaneko
authors.forename: Gwi-Gun
authors.initials: GG
authors.lastname: Park
authors.forename: Hideo
authors.initials: H
authors.lastname: Tanaka
authors.forename: Isao
authors.initials: I
authors.lastname: Kusakabe
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Kobayashi
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D006368:Helianthus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment / Q000379:methods; D020539:Sequence Analysis, Protein / Q000379:methods; D013045:Species Specificity; D013379:Substrate Specificity; D000519:alpha-Galactosidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D000519:alpha-Galactosidase