Heterologous production of lead compound terpene natural products from mushroom
Project/Area Number |
18K14342
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Research Category |
Grant-in-Aid for Early-Career Scientists
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Allocation Type | Multi-year Fund |
Review Section |
Basic Section 37020:Chemistry and chemical methodology of biomolecules-related
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Research Institution | Hokkaido University |
Principal Investigator |
Liu Chengwei 北海道大学, 理学研究院, 助教 (60773801)
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Project Period (FY) |
2018-04-01 – 2020-03-31
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Project Status |
Completed (Fiscal Year 2019)
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Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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Keywords | erinacine / mushroom / Aspergillus oryzae / heterologous expression / CRISPR/Cas9 / pleuromutilin / 生合成 / キノコ / テルペン / 異種生産 |
Outline of Final Research Achievements |
In this study, we investigated the biosynthesis of erinacin isolated from mushrooms. 1. The CRISPR / Cas9 method that achieves high transformation efficiency. Furthermore, the marker recycling method has enabled the simultaneous expression of multiple biosynthetic genes. 2. The genomic DNA required for the synthesis of erinacine is introduced into Aspergillus oryzae, 90% of introns are correctly splicing. 3. We also found an acetyltransferase outside the erinacien gene cluster, and UDP-xylose synthetic genes derived from plants. A total of 10 genes were introduced into Aspergillus oryzae, the erinacine Q was successfully produced.
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Academic Significance and Societal Importance of the Research Achievements |
キノコ由来天然物の生合成研究においては、従来cDNA配列を用いた異種発現が検討されてきた。しかし、通常の培養条件で転写されない遺伝子のcDNA配列を得ることが困難であり、研究対象が限られていた。子嚢菌と担子菌のイントロンを比較すると、長さや認識配列が非常に似ていることから、麹菌は担子菌由来の遺伝子でも正しくスプライシング可能だと着想した。本研究では、キノコが生産する神経成長因子合成促進物質エリナシンを題材として、これまでに報告例のないゲノムDNA配列を利用した異種生産を達成した。この方法はキノコ由来天然物質の汎用的な物質生産法につながると期待される。
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Report
(3 results)
Research Products
(8 results)
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[Journal Article] Efficient reconstitution of Basidiomycota diterpene erinacine gene cluster in Ascomycota host Aspergillus oryzae based on genomic DNA sequences2019
Author(s)
Liu, C., Minami, A., Ozaki, T., Wu, J., Kawagishi, H., Maruyama, J., Oikawa, H.
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Journal Title
J. Am. Chem. Soc.
Volume: 141
Issue: 39
Pages: 15519-15523
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Ascomycota Aspergillus oryzae is an efficient expression host for production of Basidiomycota terpenes using genomic DNA sequences2019
Author(s)
Shota Nagamine, Chengwei Liu, Jumpei Nishishita, Takuto Kozaki, Kaho Sogahata, Yoshiro Sato, Atsushi Minami, Taro Ozaki, Claudia Schmidt-Dannert, Jun-ichi Maruyama, Hideaki Oikawa
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Journal Title
Applied and Environmental Microbiology
Volume: 85
Issue: 15
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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