Exploratory research for innovative terpene synthases without existing motif sequences

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19065
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 38:Agricultural chemistry and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: Kuzuyama Tomohisa 東京大学, 大学院農学生命科学研究科(農学部), 教授 (30280952)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: テルペン / 生合成 / 環化酵素

Outline of Final Research Achievements

We succeeded in identifying the gene involved in the biosynthesis of a certain biologically active compound X, which has a complex cyclic structure produced by a certain microorganism. We attempted to purify the recombinant enzyme TC2, a terpene synthase that plays a central role in the production of X, and obtained a soluble enzyme. Next, the purified enzyme was incubated with Mg2+ and GGPP, which was considered as a substrate, and the reaction product was extracted with ethyl acetate, concentrated, and analyzed by GC-MS, which revealed that it was identical to the biologically active product X. These data indicate that TC2 is an enzyme that biosynthesizes biologically active product X. This is an important finding as this is the first report that a biosynthesis gene for biologically active product X has been identified.

Free Research Field

天然物化学

Academic Significance and Societal Importance of the Research Achievements

本研究課題では、複雑な化学構造をもつ生物活性物質Xの基本骨格を作り上げる生合成酵素の遺伝子を同定することに成功し、この生合成遺伝子から推定されるアミノ酸配列が、既知のテルペン合成酵素とはまったく相同性を示さない新しい酵素ファミリーであることが分かった。このような酵素の発見は、人類にとって有用な活性を示す化合物Xを大量に安価に調製するための製造方法の開発につながる成果と言える。また、この新しい酵素ファミリーに属するテルペン合成酵素をゲノムデータベースからさらに発掘して機能解析を行うことで、人類がこれまでに手にしたことのない新たな有用なテルペン化合物の発見につながる点でも重要な成果と言える。