Cloning and Homologous Expression of Novel Lignin Peroxidase Genes in the White-Rot Fungus Phanerochaete sordida YK-624
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概要
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Two genes, encoding YK-LiP1 and YK-LiP2, were cloned from the white-rot fungus Phanerochaete sordida YK-624, and a homologous expression system for the gene was constructed. Two full-length cDNAs (ylpA and ylpB) were isolated by degenerate RT-PCR and RACE-PCR. The results of N-terminal amino acid sequence analysis of native YK-LiP1 and YK-LiP2 showed that ylpA and ylpB coded for YK-LiP2 and YK-LiP1 respectively. The promoter of glyceraldehyde-3-phosphate dehydrogenase cloned from P. sordida YK-624 (PsGPD) was used to drive the expression of ylpA. Expression vector pGPD-g-ylpA was transformed into a P. sordida YK-624 uracil auxotrophic mutant, UV-64. The YlpA protein was secreted in active form by the transformants after 4 d of growth in a medium containing an excessive nitrogen source, whereas endogenous YK-LiP1 and YK-LiP2 were not produced. The physical and catalytic properties of the purified YlpA protein were very similar to those of YK-LiP2. These results suggest that homologous expression of recombinant YK-LiP2 was successful.
- 社団法人 日本農芸化学会の論文
著者
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Kawagishi Hirokazu
Department Of Applied Biological Chemistry Faculty Of Agriculture Shizuoka University
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HIRAI Hirofumi
Department of Forest Resources Science, Shizuoka University
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NISHIDA Tomoaki
Department of Forest Resources Science, Shizuoka University
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Sugiura Tatsuki
Department of Bioscience, Graduate School of Science and Technology, Shizuoka University
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Kimura Toshiyuki
National Institute Of Information And Communications Technology
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Kimura Toshiyuki
Japan National Agricultural Res. Center For Tohoku Region National Agriculture And Food Res. Organiz
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Yamagishi Kenji
National Agricultural Research Center for the Tohoku Region, National Agriculture and Food Research
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Kimura Toshiyuki
National Agricultural Research Center For The Tohoku Region National Agriculture And Food Research O
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