Molecular Cloning and Expression of Two Novel β-N-Acetylglucosaminidases from Silkworm Bombyx mori
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概要
- 論文の詳細を見る
β-N-Acetylglucosaminidase is a major glycosidase involved in several physiological processes, such as fertilization, metamorphosis, glycoconjugate degradation, and glycoprotein biosynthesis in insects. A search using the Bombyx mori cDNA database revealed the existence of two putative β-N-acetylglucosaminidase genes. Their full-length cDNAs were cloned by rapid amplification of cDNA ends and polymerase chain reaction using specific primers, and named BmGlcNAcase1 and BmGlcNAcase2. A BLAST search revealed that BmGlcNAcase1 and BmGlcNAcase2 are homologous to a β-subunit homolog encoded by Drosophila melanogaster HEXO2 and the Spodoptera frugiperda β-N-acetylglucosaminidase gene respectively. The recombinant proteins of BmGlcNAcase1 and BmGlcNAcase2 without putative transmembrane domains were expressed in the yeast Pichia pastoris. Both enzymes showed broad substrate specificity, and cleaved terminal N-acetylglucosamine residues from the α-3 and α-6 branches of a biantennary N-glycan substrate, and also hydrolyzed chitotriose to chitobiose.
- 社団法人 日本農芸化学会の論文
- 2007-07-23
著者
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MITA Kazuei
National Institute of Agrobiological Sciences
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Fujiyama Kazuhito
The International Center for Biotechnology, Osaka University
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SEKI Tatsuji
The International Center for Biotechnology, Osaka University
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Ishiyama Seiji
Research Institute of Biological Science, Katakura industries CO., LTD.
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Mita Kazuhide
National Institute of Agrobiological Sciences
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Tamura Toshiki
National Institute of Agrobiological Sciences
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Fujiyama Kazuhito
International Center For Biotechnology Osaka Univ.
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Fujiyama Kazuhito
International Center For Biotechnology Osaka University
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Seki Tatsuji
The International Center For Biotechnology Osaka University
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Usami Akihiro
Research Institute Of Biological Science Katakura Industries Co. Ltd.
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Okada Takahiro
International Center For Biotechnology Osaka University:(present Office)division Of Molecular Cell B
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Ishiyama Seiji
Research Institute Of Biological Science Katakura Industries Co. Ltd.
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Sezutsu Hideki
National Institute of Agrobiological Sciences
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Fujiyama Kazuhito
The International Center For Biotechnology Osaka Univ. 2-1 Yamada-oka Suita-shi Osaka 565-0871 Jpn
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Mita Kazuei
National Institute Of Agrobiological Science
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OKADA Takahiro
The International Center for Biotechnology, Osaka University
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ISHIYAMA Seiji
Department of Research Development, Katakura Industries Co., Ltd.
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USAMI Akihiro
Department of Research Development, Katakura Industries Co., Ltd.
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Fujiyama Kazuhito
The International Center For Biotechnology Osaka Univ. 2-1 Yamada-oka Suita Osaka 565-0871 Jpn
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Tamura Toshiki
Division Of Insect Sciences National Institute Of Agrobiological Sciences
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Sezutsu Hideki
Transgenic Silkworm Research Unit Genetically Modified Organism Research Center National Institute Of Agrobiological Sciences
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