Molecular Cloning and Expression in Pichia pastoris of a Irpex lacteus Exo-β-(1→3)-galactanase Gene
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概要
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A gene encoding exo-β-(1→3)-galactanase from Irpex lacteus was cloned by reverse transcriptase-PCR. The deduced amino acid sequence showed high similarity with exo-β-(1→3)-galactanases from other sources. The molecular mass of the mature form was calculated to be 45,520 Da. The gene product expressed in Pichia pastoris specifically hydrolyzed β-(1→3)-galactooligosaccharides, as did other exo-β-(1→3)-galactanases. The recombinant enzyme showed high activity toward arabinogalactan-proteins (AGPs) from radish as well as β-(1→3)-galactan. Product analysis revealed that the enzyme released β-(1→6)-galactobiose, β-(1→6)-galactotriose, and α-L-arabinofuranosyl-(1→3)-β-galactosyl-(1→6)-galactose together with Gal from β-(1→3)-galactans attached with and without β-(1→6)-galactosyl branches prepared from acacia gum. These results indicate that the exo-β-(1→3)-galactanase from I. lacteus efficiently hydrolyzes β-(1→3)-galactan main chains of AGPs by bypassing β-(1→6)-galactosyl side chains.
- 社団法人 日本農芸化学会の論文
- 2009-10-23
著者
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Tsumuraya Yoichi
Division of Life Science, Graduate School of Science and Engineering, Saitama University
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KANEKO Satoshi
Food Biotechnology Division, National Food Research Institute
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Kaneko Satoshi
Food Biotechnology Div. National Food Res. Inst.
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Tsumuraya Yoichi
Division Of Life Science Graduate School Of Science And Engineering Saitama University
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Tsumuraya Yoichi
Graduate School Of Science And Engineering Saitama University
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Tsumuraya Yoichi
Div. Of Life Sci. Graduate School Of Sci. And Engineering Saitama Univ.
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ICHINOSE Hitomi
Food Biotechnology Division, National Food Research Institute
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Kotake Toshihisa
Division Of Life Science Graduate School Of Science And Engineering Saitama University
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Okabe Kohei
Division Of Life Science Graduate School Of Science And Engineering Saitama University
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KITAZAWA Kiminari
Division of Life Science, Graduate School of Science and Engineering, Saitama University
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TAKATA Ryohei
Division of Life Science, Graduate School of Science and Engineering, Saitama University
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Kitazawa Kiminari
Division Of Life Science Graduate School Of Science And Engineering Saitama University
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Takata Ryohei
Division Of Life Science Graduate School Of Science And Engineering Saitama University
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Ichinose Hitomi
National Food Res. Inst.
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