Purification and structural analysis of volatile sesquiterpenes produced by Escherichia coli carrying unidentified terpene synthase genes from edible plants of the family Araliaceae

Author:

Shindo Kazutoshi1,Hattan Jun-ichiro2,Kato Mariko1,Sato Miho1,Ito Tomoko1,Shibuya Yurika1,Watanabe Arisa1,Sugiyama Maki1,Nakamura Yuri1,Misawa Norihiko2

Affiliation:

1. Department of Food and Nutrition, Japan Women’s University, Tokyo, Japan

2. Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Ishikawa, Japan

Abstract

Abstract A simple method to purify volatile sesquiterpenes from recombinant Escherichia coli was developed using the cells that carried known sesquiterpene synthase (Tps) genes ZzZss2 (ZSS2) and ZoTps1. This method was applied for the purification and structural analyses of volatile sesquiterpenes produced by E. coli cells that carried unidentified Tps genes, which were isolated from the Aralia-genus edible plants belonging to the family Araliaceae. Recombinant cells carrying each Tps gene were cultured in the two-layer medium (n-octane/TB medium), and volatile sesquiterpenes trapped in n-octane were purified through two-phase partition, silica gel column chromatography, and reversed-phase preparative high-performance liquid chromatography, if necessary. Further, their structures were confirmed by nuclear magnetic resonance, [α]D, and gas chromatography–mass spectrometry analyses. Herein, the products of E. coli cells that carried two Tps gene (named AcTps1 and AcTps2) in Araria cordata “Udo” and a Tps gene (named AeTps1) in Aralia elata “Taranoki” were studied resulting in identifying functionalities of these cryptic Tps genes.

Publisher

Oxford University Press (OUP)

Subject

Organic Chemistry,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Biochemistry,Analytical Chemistry,Biotechnology

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