Identification and functional analysis of a novel terpene cyclase that enables the creation of unnatural meroterpenoid

• Project/Area Number: 25850079
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Bioorganic chemistry
• Research Institution: Niigata University
• Principal Investigator: NAKANO CHIAKI 新潟大学, 自然科学系, 助教 (70620376)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: テルペン環化酵素 / メロテルペノイド / 生合成 / 放線菌 / BE-40644

Outline of Final Research Achievements

To identify the novel terpene cyclase responsible for synthesis of BE-40644, in vivo analysis of BE-40644 biosynthetic gene cluster was performed. The putative terpene cyclase orf15 was deleted from pKU1021-orf8-15 and the resultant plasmid was transformed into Streptomyces avermitilis. Moreover, the E44A mutant of orf15 and the deletion mutant of putative prenyltransferase orf14 were constructed. The orf15 deletion mutant gave no product. On the other hand, a new product was accumulated in the E44A mutant and the orf14 deletion mutant. Incubation of ORF15 with substrate analogues such as farnesyl hydroquinone gave no product.