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Plant and Cell Physiology Advance Access first published online on November 3, 2009
This version published online on November 4, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp156
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Metabolic Engineering of Lignan Biosynthesis in Forsythia Cell Culture

Hyun Jung Kim1, Eiichiro Ono2, Kinuyo Morimoto1, Tohru Yamagaki1, Atsushi Okazawa3, Akio Kobayashi3 and Honoo Satake1

1Suntory Institute for Bioorganic Research, 1-1-1 Wakayamadai, Shimamoto, Mishima, Osaka, 618-8503 JAPAN
2Core Research Group, R&D Planning Division, Suntory Holdings Ltd, Suntory Research Center, 1-1-1 Wakayamadai, Shimamoto, Mishima, Osaka 618-8503, JAPAN
3Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita, Osaka 565-0871, JAPAN

Corresponding Author: Honoo Satake Address: Suntory Institute for Bioorganic Research, 1-1-1 Wakayamadai, Shimamoto, Mishima, Osaka, 618-8503 JAPAN E-mail: satake{at}sunbor.or.jp Tel: +81-75-962-6092 Fax: +81-75-962-2115


  Abstract

Lignans are a large class of secondary metabolites in plants, with numerous biological effects in mammals, including anti-tumor and anti-oxidant activities. Sesamin, the most abundant furofuran-class lignan in sesame seeds (Sesamum plants), is produced by the cytochrome P450 enzyme CYP81Q1 from the precursor lignan, pinoresinol. In contrast, Forsythia plants produce dibenzylbutyrolactone-class lignans, such as matairesinol, from pinoresinol via the catalysis of pinoresinol/lariciresinol reductase (PLR) and secoisolariciresinol dehydrogenase. Here we present the engineering of lignan biosynthesis in Forsythia cell suspension cultures for the development of an efficient production method of beneficial lignans. A suspension cell culture prepared from leaves of Forsythia koreana produced lignans, mainly pinoresinol and matairesinol glucosides, at levels comparable to that obtained from the leaves. In an attempt to increase the pinoresinol content in Forsythia, we generated a transgenic cell line overexpressing an RNA interference (RNAi) construct of PLR (PLR-RNAi). Down-regulation of PLR expression led to a complete loss of matairesinol and an accumulation of approximately 20-fold pinoresinol in its glucoside form in comparison with the non-transformant. Moreover, the Forsythia transgenic cells co-expressing CYP81Q1 and PLR-RNAi exhibited production of sesamin as well as accumulation of pinoresinol glucoside. These data suggest Forsythia cell suspension to be a promising tool for the engineering of lignan production. To the best of our knowledge, this is the first report on transgenic production of an exogenous lignan in a plant species.

Keywords: Forsythia koreana - Lignan - Metabolic engineering - Sesamin - Sesamum indicum - Suspension cell culture.

(Received August 11, 2009; Accepted October 26, 2009)
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