Development of a new method for the quantification of isoprenoid intermediates and analysis of the localization of the isoprenoid metabolic pathways in chloroplasts

2014 Fiscal Year Final Research Report

• Project/Area Number: 26650088
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Plant molecular biology/Plant physiology
• Research Institution: Hokkaido University
• Principal Investigator: TANAKA Ryouichi 北海道大学, 低温科学研究所, 准教授 (20311516)
• Project Period (FY): 2014-04-01 – 2015-03-31
• Keywords: 葉緑体

Outline of Final Research Achievements

We aimed at the development of a method to quantify geranylgeranyl-diphosphate (GGDP) and phytyl-diphosphate (PDP) in this study. First, we have tested whether geranylgeranyl transferase can conjugate GGDP and PDP to a dansylated peptide, which can be easily detected by a liquid chromatography system according to the method of Tong et al. 2013. However, this method only detect GGDP-conjugated peptide. Then, we attempted to use chlorophyll synthase activity to detect GGDP- or PDP-conjugate to chlorophyllide. However, we failed to overexpress this enzyme by an E. coli system, cultured insect cells (Sf9), and the MembraneMax cell-free expression system.

Free Research Field

植物生理学