Cloning of a Novel Tyrosinase-Encoding Gene (melB) from Aspergillus oryzae and Its Overexpression in Solid-State Culture (Rice Koji)(BREWING AND FOOD TECHNOLOGY)
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概要
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We have cloned a novel tyrosinase-encoding gene (melB) specifically expressed in solid-state culture of Aspergillus oryzae. A tyrosinase-encoding gene (melO) from A. oryzae was already cloned and the protein structures of its catalytic and copper binding domains were investigated. However, our recent results revealed that the melO gene was highly expressed in submerged culture but not in solid-state culture. Because tyrosinase activity was also detected in solid-state culture, we assumed that another tyrosinase gene other than melO is expressed in solid-state culture. Another tyrosinase gene was screened using the expressed sequence tag (EST) library. One redundant cDNA clone homologous with the tyrosinase gene was found in the collection of wheat bran culture. Northern blot analysis revealed that the gene corresponding to the cDNA clone was specifically expressed in solid-state culture (koji making), but not in submerged culture. Molecular cloning showed that the gene carried six exons interrupted by five introns and had an open reading frame encoding 616 amino acid residues. This gene was designated as melB. The deduced amino acid sequence of the gene had weak homology (24%-33%) with MelO and other fungal tyrosinases but the sequences of the copper binding domains were highly conserved. When the melB gene was expressed under the control of the glaB promoter in solid-state culture, tyrosinase activity was markedly enhanced and the culture mass was browned with the melanization by MelB tyrosinase. These results indicated that the melB gene encodes a novel tyrosinase associated with melanization in solid-state culture.
- 社団法人日本生物工学会の論文
- 2004-06-25
著者
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Akao Takeshi
National Research Institute of Brewing
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Hata Yoji
Research Institute, Gekkeikan Sake Co. Ltd.
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Hata Yoji
Res. Inst. Gekkeikan Sake Co. Ltd. 101 Shimotoba-koyanagi-cho Fushimi-ku Kyoto 612-8385 Jpn
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Ishida H
Research Institute Gekkeikan Sake Co. Ltd.
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Ishida Hiroki
Research Institute Gekkeikan Sake Co. Ltd.
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Abe Yasuhisa
Res. Inst. Gekkeikan Sake Co. Ltd.
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Abe Yasuhisa
Research Institute For Fundamental Physics Kyoto University : Centre De Recherches Nucleaires
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Abe Yasuhisa
Research Institute For Fundamental Physics Kyoto University
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KAWATO Akitsugu
Research Institute, Gekkeikan Sake Co., Ltd.
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AKITA Osamu
National Research Institute of Brewing (NRIB)
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Obata Hiroshi
Research Institute Gekkeikan Sake Co. Ltd.
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ICHISHIMA EIJI
Department of Bioengineering, Graduate School of Engineering, Soka University
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Ichishima Eiji
Department Of Bioengineering Graduate School Of Engineering Soka University
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Ichishima E
Department Of Bioengineering Graduate School Of Engineering Soka University
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Ichishima Eiji
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Hata Yoji
Research Institite Of Gekkeikan Gekkeikan Sake Co. Ltd.
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Abe Yasushisa
Research Institite Of Gekkeikan Gekkeikan Sake Co. Ltd.
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Akita O
Fac. Of Human Life Sciences Dep. Of Food And Health Sciences Jissen Women's Univ.
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Akita Osamu
National Research Institute Of Brewing
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Kawato A
Research Institute Gekkeikan Sake Co. Ltd.
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Kawato Akitsugu
Research Institute Gekkeikan Sake Co. Ltd.
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Abe Yasuhisa
Research Institute Gekkeikan Sake Co. Ltd.
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Ichikawa Eiji
Gekkeikan Research Institute Gekkeikan Sake Co. Ltd.
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Abe Yasuhisa
Research Center For Nuclear Physics Osaka University
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