Engineering of plant-specific polyketide scaffold synthetic enzymes to produce structural diversity of compounds

2016 Fiscal Year Final Research Report

• Project/Area Number: 26282210
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: University of Toyama
• Principal Investigator: Morita Hiroyuki 富山大学, 和漢医薬学総合研究所, 教授 (20416663)
• Co-Investigator(Renkei-kenkyūsha): MATSUI Takashi 富山大学, 和漢医薬学総合研究所, 助教 (30463582) ; KODAMA Takeshi 富山大学, 和漢医薬学総合研究所, 特命助教 (40710207)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ポリケタイド / 酵素工学 / X線結晶構造解析

Outline of Final Research Achievements

In order to produce structural diversity of compounds, combinatorial biosyntheses of the polyketide cyclase, olivetolic acid cyclase (OAC) and the type III polyketide synthase, octaketide synthase from Aloe arborescens using structurally distinct various CoA thioesters as the substrates were carried out. As a result, we found that the co-incubation of the enzymes using hexanoyl-CoA and malonyl-CoA as the substrates led to a production of novel naphthalene. Furthermore, we solved OAC apo and OAC-olivetolic acid binary complex structures at 1.4 angstrom and 1.7 angstrom resolutions, respectively. The crystallographic and site-directed mutagenesis studies of OAC revealed that the enzyme possesses the pentyl-binding pocket that plays an important role to bind the pentyl moiety of the substrate and employs Tyr72 and His78 as the catalytic residues.

Free Research Field

天然物化学