Ishida Hiroki, Moritani Toshiyasu, Hata Yoji, Kawato Akitsugu, Suginami Koji, Abe Yasuhisa, Imayasu Satoshi
Research Institute, Gekkeikan Sake Co. Ltd, Kyoto, Japan.
Biosci Biotechnol Biochem. 2002 May;66(5):1002-8. doi: 10.1271/bbb.66.1002.
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.
用固定有稻曲霉铁载体之一的铁曲霉素（Fcy）的柱色谱法，从米曲霉细胞裂解物中纯化出一种蛋白质。它仅在缺铁培养物中产生，通过SDS-PAGE估计其分子量为35000。分析了通过赖氨酰内肽酶消化获得的该蛋白质的两个内部氨基酸序列。分子克隆表明，它编码由4个内含子隔开的310个推定氨基酸残基，并被命名为fleA。它与橙黄网柄菌（AAL）的岩藻糖特异性凝集素编码基因显示出约26%的相似性。该基因在米曲霉深层培养中在melO启动子的控制下过表达。fleA基因产物对兔红细胞显示出血凝活性。单糖的血凝抑制试验表明，这种凝集素特异性结合L-岩藻糖，并与甘露糖和N-乙酰神经氨酸弱反应。
