Studies on Triterpene Production using biosyntetic genes

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K08310
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Natural medicines
• Research Institution: Nigata University of Phermacy and Applied Life Sciences
• Principal Investigator: Shibuya Masaaki 新潟薬科大学, 薬学部, 教授 (50170923)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: トリテルペン / ソヤサポゲノールB / グルクロン酸転移酵素

Outline of Final Research Achievements

I supposed that it is indispensable to excrete the product from yeast cells by glycosylation in order to accomplish the high production of triterpene by hetelorogous gene expression in transformed yeast. For this reason, I started cloning of triterpene glycosyltransferase (UDP-glucuronate : soyasapogenol B glucuronosyltransferase). In order to clone this transferase gene, in vivo screening assay using the yeast which was transformed with UDP-glucose dehydrogenase gene, was used. UDP-glucose dehydrogenase gene was successfully cloned from soybean, and introduced into yeast expression plasmid pESC. Resulting plasmid pESC-DH was obtained, and candidate triterpene glycosyltransferase genes was cloned into pESC-DH. During the research period, activities of 20 candidate genes were investigated using the transformed yeast with the constructed plasmid, but no active clone was found. Investing the activity of remained 11 candidate genes was continued.

Free Research Field

天然物化学

Academic Significance and Societal Importance of the Research Achievements

本研究の目的としたトリテルペン第一配糖化酵素遺伝子をクローニングすることができれば、トリテルペンサポニンの酵母による大量生産が可能となると期待される。理研のグループもトリテルペンサポニンであるグリチルリチンの酵母による大量生産系の構築を進めている。グリチルリチンは医薬品として有用であり、本研究が成功すれば、理研グループのグリチルリチンの大量生産系の構築に寄与するものと思われる。