title: Molecular cloning of cDNA for trehalase from the European honeybee, Apis mellifera L., and its heterologous expression in Pichia pastoris.
abstract: cDNA encoding the bound type trehalase of the European honeybee was cloned. The cDNA (3,001 bp) contained the long 5' untranslated region (UTR) of 869 bp, and the 3' UTR of 251 bp including a poly(A) tail, and the open reading frame of 1,881 bp consisting of 626 amino acid residues. The Mr of the mature enzyme comprised of 591 amino acids, excluded a signal sequence of 35 amino acid residues, was 69,177. Six peptide sequences analyzed were all found in the deduced amino acid sequence. The amino acid sequence exhibited high identity with trehalases belonging to glycoside hydrolase family 37. A putative transmembrane region similar to trehalase-2 of the silkworm was found in the C-terminal amino acid sequence. Recombinant enzyme of the trehalase was expressed in the methylotrophic yeast Pichia pastoris as host, and displayed properties identical to those of the native enzyme except for higher sugar chain contents. This is the first report of heterologous expression of insect trehalase.
authors.affiliation: Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo, 060-8589, Japan.
authors.forename: Jin-Ha
authors.initials: JH
authors.lastname: Lee
authors.forename: Saori
authors.initials: S
authors.lastname: Saito
authors.forename: Haruhide
authors.initials: H
authors.lastname: Mori
authors.forename: Mamoru
authors.initials: M
authors.lastname: Nishimoto
authors.forename: Masayuki
authors.initials: M
authors.lastname: Okuyama
authors.forename: Doman
authors.initials: D
authors.lastname: Kim
authors.forename: Jintanart
authors.initials: J
authors.lastname: Wongchawalit
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 / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014198:Trehalase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*