Molecular cloning and overexpression of fleA gene encoding a fucose-specific lectin of Aspergillus oryzae

被引：37

作者：

Ishida, H ^{[1
]}

Moritani, T ^{[1
]}

Hata, Y ^{[1
]}

Kawato, A ^{[1
]}

Suginami, K ^{[1
]}

Abe, Y ^{[1
]}

Imayasu, S ^{[1
]}

机构：

[1] Gekkeikan Sake Co Ltd, Inst Res, Fushimi Ku, Kyoto 6128361, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2002年 / 66卷 / 05期

关键词：

fucose-specific lectin; Aspergillus oryzae; fleA; melO promoter; ferrichrysin;

D O I：

10.1271/bbb.66.1002

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 fucosespecific 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.

引用

页码：1002 / 1008

页数：7

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