Characterization of amylolytic enzyme overproducing mutant of <i>Aspergillus luchuensis</i> obtained by ion beam mutagenesis
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- Kojo Toshihiro
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Kadooka Chihiro
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Komohara Marisa
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Onitsuka Shiori
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Tanimura Miya
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Muroi Yukiko
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Kurazono Shugo
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Shiraishi Yohei
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University Bio’c, Co., Ltd
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- Oda Ken
- National Research Institute of Brewing
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- Iwashita Kazuhiro
- National Research Institute of Brewing
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- Onoue Masahira
- Natural Science Centre for Research and Education, Kagoshima University
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- Okutsu Kayu
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Yoshizaki Yumiko
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Takamine Kazunori
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Futagami Taiki
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
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- Mori Kazuki
- Computational Bio-Big Data Open Innovation Laboratory, National Institute of Advanced Industrial Science and Technology
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- Tamaki Hisanori
- Education and Research Centre for Fermentation Studies, Faculty of Agriculture, Kagoshima University
抄録
<p>Aspergillus luchuensis is a kuro (black) koji fungus that has been used as a starch degrader for the awamori- and shochu-making industries in Japan. In this study, we investigated the effect of ion beam irradiation on A. luchuensis RIB2601 and obtained a high starch-degrading mutant strain U1. Strain U1 showed reduced growth rate, whereas it showed higher α-amylase, glucoamylase, and α-glucosidase activities on a mycelial mass basis than the wild type (wt) strain both on agar plates and in rice koji. In addition, strain U1 showed higher N-acetylglucosamine content in the cell wall and higher sensitivity to calcofluor white, suggesting a deficiency in cell wall composition. Interestingly, produced protein showed higher expression of acid-labile α-amylase (AmyA) and glucoamylase (GlaA) in strain U1, although real-time RT-PCR indicated no significant change in the transcription of the amyA or glaA gene. These results suggested that the high amylolytic activity of strain U1 is attributable to a high AmyA and GlaA production level, but the elevated production is not due to transcriptional regulation of the corresponding genes. Furthermore, RNA-seq analysis indicated that strain U1 shows transcriptional changes in at least 604 genes related to oxidation-reduction, transport, and glucosamine-containing compound metabolic processes, which may be involved in the deficient cell wall composition of strain U1.</p>
収録刊行物
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- The Journal of General and Applied Microbiology
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The Journal of General and Applied Microbiology 63 (6), 339-346, 2017
公益財団法人 応用微生物学・分子細胞生物学研究奨励会