Development of efficient gene expression systems for secondary metabolite production in filamentous fungi

2014 Fiscal Year Final Research Report

• Project Area: Biosynthetic machinery: Deciphering and regulating the system for bioactive metabolite diversification
• Project/Area Number: 22108007
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Tohoku University
• Principal Investigator: GOMI Katsuya 東北大学, (連合)農学研究科(研究院), 教授 (60302197)
• Co-Investigator(Kenkyū-buntansha): MACHIDA Masayuki 産業技術総合研究所, 生物プロセス部門, チームリーダー (30358006)
• Co-Investigator(Renkei-kenkyūsha): SHINTANI Takahiro 東北大学, 大学院農学研究科, 准教授 (70374973) ; KOIKE Hideaki 産業技術総合研究所, 生物プロセス部門, 主任研究員 (90344118)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Keywords: 二次代謝化合物 / 多重遺伝子導入 / 遺伝子破壊 / マーカーリサイクリング / 生合成酵素 / 細胞内局在 / トランスポーター / 物質生産

Outline of Final Research Achievements

We developed a self-excising Cre/loxP-mediated marker recycling system with mutated lox sequences to introduce a number of biosynthetic genes into Aspergillus oryzae. By using the marker recycling system constructed, hyperproduction of kojic acid could be achieved in A. oryzae by the introduction of the two genes that each encodes oxidoreductase and transporter. Deletion of the transporter resulted in a significant decrease in the yield of kojic acid compared to the wild type strain, indicating that overexpression of the transporter gene is highly effective in improving the production yield of secondary metabolites. In addition, expression vectors with multiple expression cassettes consisting of the amyB promoter and terminator were also constructed. We examined subcellular localization of the enzyme proteins encoded by a gene cluster involved in aphidicolin biosynthesis from Phoma betae, which were expressed as GFP-fusion proteins in A. oryzae.

Free Research Field

応用微生物学