Secretion of Mono- and Diacylglycerol Lipase from Penicillium camembertii U-150 by Saccharomyces cerevisiae and Site-directed Mutagenesis of the Putative Catalytic Sites of the Lipase
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概要
- 論文の詳細を見る
Yeast cells carrying intronless mono- and diacylglycerol lipase (MDGL) genes, constructed by recombination of the genomic gene and cDNA, secreted MDGL into the culture supernatant. Most of the yeast MDGL were extensively glycoslated while they had a similar glyceride specificity to that of native MDGL. Site-directed mutagenesis was used to directly confirm the involvements in enzyme activity of the presumptive amino acid residues to form the catalytic center of MDGL. These residues were conserved in the primary structure alignment of a lipase family from filamentous fungi. Mutant lipase proteins in which Ser^<83>, Ser^<145>, or His<259> was replaced with glycine were secreted by yeast transformants as inactive proteins. Mutant proteins replacing Asp^<199> with glycine or asparagine were not detected in the culture supernatant. Replacing other two highly conserved aspartic acids (at positions 232 and 243) with glycine did not render the enzyme inactive. These results indicate that Ser^<83>, Ser^<145>, and His^<259> in MDGL, are essential to enzyme activity. Asp^<199> is also likely to be involved.
- 社団法人日本農芸化学会の論文
- 1992-02-23
著者
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YAMAGUCHI Shinjiro
RIKEN Plant Science Center
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Yamaguchi Shinjiro
Plant Science Center Riken
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Yamaguchi Shinjiro
Frontier Research Program Riken
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Takeuchi Kazuyuki
Research And Development Division Amano Prarmaceutical Co. Ltd.
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Takeuchi Kazuyuki
Tsukuba Research Laboratories Amano Pharmaceutical Co. Ltd.
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MASE TAMIO
Research and Development Division, Amano Pharmaceutical Co. Ltd.,
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Mase T
Process Research Process R&d Laboratories For Technology Development Banyu Pharmaceutical Co. Lt
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Mase Tamio
Research And Development Division Amano Pharmaceutical Co. Ltd.
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Yamaguchi S
Riken Plant Science Center
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Mase T
Amano Pharmaceutical Co. Ltd. Aichi Jpn
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YAMAGUCHI Shotaro
Tsukuba Research Laboratories, Amano Pharmaceutical Co., Ltd.
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