Elucidation and controlment of diverse structure in the biosynthesis of indole diterpene

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K16636
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Hokkaido University
• Principal Investigator: Liu Chengwei 北海道大学, 理学研究院, 助教 (60773801)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: biosynthesis / indole diterpene / Indole sesquiterpene / shearinine / sespendole

Outline of Final Research Achievements

Indole terpenes are a large class of fungal natural product. In this study, we used Aspergillus oryzea heterologous expression system to express shearinine and sespendole biosynthetic genes. ①The late-stage biosynthetic pathway of the shearinine involving four enzymatic reaction was elucidated by heterologous expression in A. oryzae. When incubated this transformants with the corresponding intermediates, we found that shearinine D was produced. In vitro enzymatic reaction with a recombinant protein, we elucidated the key oxidative cyclization, forming a characteristic 6/5 bicyclic of shearinine by flavoprotein oxidase. ② Sespendole biosynthetic gene cluster were identified. The biosynthetic pathway that involves genes encoding six enzymes was elucidated through heterologous expression of spd genes in A. oryzae. Thereby, revealing the mechanism underlying the characteristic modification on the indole ring, catalyzed by a set of prenyltransferase (SpdE)/cytochrome P450 (SpdJ) enzymes.

Free Research Field

天然物化学