title: Molecular cloning and overexpression of fleA gene encoding a fucose-specific lectin of Aspergillus oryzae.
abstract: A protein from the cell lysate of Aspergillus oryzae was purified by column chromatography immobilized with a ferrichrysin (Fcy), which is one of the siderophores of A. oryzae. It is produced only in an iron-deficient culture and its molecular weight is estimated as 35,000 by SDS-PAGE. Two internal amino acid sequences of the protein obtained by lysylendopeptidase digestion were analyzed. Molecular cloning shows that it encodes 310 putative amino acid residues separated by 4 introns and is designated as fleA. It shows approximately 26% similarity with the gene encoding a fucose-specific lectin of Aleuria aurantia (AAL). The gene was overexpressed under control of the melO promoter in a submerged culture of A. oryzae. The fleA gene product showed hemagglutination activity against rabbit erythrocytes. A hemagglutination inhibition assay of monosaccharides showed that this lectin specifically binds to L-fucose and weakly reacts with mannose and N-acetyl-neuraminic acid.
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd, Kyoto, Japan. h-ishida@gekkeikan.co.jp
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ishida
authors.forename: Toshiyasu
authors.initials: T
authors.lastname: Moritani
authors.forename: Yoji
authors.initials: Y
authors.lastname: Hata
authors.forename: Akitsugu
authors.initials: A
authors.lastname: Kawato
authors.forename: Koji
authors.initials: K
authors.lastname: Suginami
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Abe
authors.forename: Satoshi
authors.initials: S
authors.lastname: Imayasu
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000235:genetics*; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004591:Electrophoresis, Polyacrylamide Gel; D005643:Fucose / Q000378:metabolism*; D037102:Lectins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
chemical_list: D004271:DNA, Fungal; D037102:Lectins; D005643:Fucose