title: Molecular cloning of cDNAs and genes for three alpha-glucosidases from European honeybees, Apis mellifera L., and heterologous production of recombinant enzymes in Pichia pastoris.
abstract: cDNAs encoding three alpha-glucosidases (HBGases I, II, and III) from European honeybees, Apis mellifera, were cloned and sequenced, two of which were expressed in Pichia pastoris. The cDNAs for HBGases I, II, and III were 1,986, 1,910, and 1,915 bp in length, and included ORFs of 1,767, 1,743, and 1,704 bp encoding polypeptides comprised of 588, 580, and 567 amino acid residues, respectively. The deduced proteins of HBGases I, II, and III contained 18, 14, and 8 putative N-linked glycosylation sites, respectively, but at least 2 sites in HBGase II were unmodified by N-linked oligosaccharide. In spite of remarkable differences in the substrate specificities of the three HBGases, high homologies (38-44% identity) were found in the deduced amino acid sequences. In addition, three genomic DNAs, of 13,325, 2,759, and 27,643 bp, encoding HBGases I, II, and III, respectively, were isolated from honeybees, and the sequences were analyzed. The gene of HBGase I was found to be composed of 8 exons and 7 introns. The gene of HBGase II was not divided by intron. The gene of HBGase III was confirmed to be made up of 9 exons and 8 introns, and to be located in the region upstream the gene of HBGase I.
authors.affiliation: Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Nishimoto
authors.forename: Haruhide
authors.initials: H
authors.lastname: Mori
authors.forename: Tsuneharu
authors.initials: T
authors.lastname: Moteki
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Takamura
authors.forename: Gaku
authors.initials: G
authors.lastname: Iwai
authors.forename: Yu
authors.initials: Y
authors.lastname: Miyaguchi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Okuyama
authors.forename: Jintanart
authors.initials: J
authors.lastname: Wongchawalit
authors.forename: Rudee
authors.initials: R
authors.lastname: Surarit
authors.forename: Jisnuson
authors.initials: J
authors.lastname: Svasti
authors.forename: Atsuo
authors.initials: A
authors.lastname: Kimura
authors.forename: Seiya
authors.initials: S
authors.lastname: Chiba
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D010843:Pichia; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D000520:alpha-Glucosidases / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D000520:alpha-Glucosidases