Revisiting anabasine biosynthesis in tobacco hairy roots expressing plant lysine decarboxylase gene by using ¹⁵N-labeled lysine

• Bunsupa Somnuk

  Graduate School of Pharmaceutical Sciences, Chiba University

• Komastsu Kana

  Graduate School of Pharmaceutical Sciences, Chiba University

• Nakabayashi Ryo

  RIKEN Center for Sustainable Resource Science

• Saito Kazuki

  Graduate School of Pharmaceutical Sciences, Chiba University RIKEN Center for Sustainable Resource Science

• Yamazaki Mami

  Graduate School of Pharmaceutical Sciences, Chiba University

Bibliographic Information

Other Title

• Revisiting anabasine biosynthesis in tobacco hairy roots expressing plant lysine decarboxylase gene by using ¹⁵N-labeled lysine

Abstract

Anabasine is an alkaloid found in a small number of Nicotiana species. The components of the anabasine biosynthetic pathway have yet to be identified. Here, we report the reinvestigation of biosynthetic pathways of anabasine and related tobacco alkaloids in genetically engineered cells. Hairy roots of N. tabacum harboring a lysine/ornithine decarboxylase gene from Lupinus angustifolius (La-L/ODC) were fed with labeled [ε-¹⁵N]- or [α-¹⁵N]-L-lysine. Relative to the unfed control, feeding of labeled ¹⁵N-L-lysine greatly enhanced anabasine levels 13.5-fold in La-L/ODC-expressing line compared to 5.3-fold in the control line, suggesting that both LDC activity and substrate supplied are important factors for the efficient production of anabasine. GUS-expressing line showed preferential incorporation of [ε-¹⁵N]-L-lysine into anabasine, indicating the main biosynthetic pathway of Δ¹-piperideine intermediate in tobacco is asymmetrically processes. In contrast, the expression of La-L/ODC showed the symmetric labeling of ¹⁵N atom into anabasine, implying the occurrence of free cadaverine, which is produced by La-L/ODC enzyme, during the biosynthesis of Δ¹-piperideine intermediate. No considerable incorporation of ¹⁵N into other tobacco alkaloids such as, nicotine, anatabine, and anatalline, was detected. Detailed analysis using ultra-high resolution mass spectrometry indicated that two ¹⁵N atoms were incorporated into anabasine in La-L/ODC-expressing lines after feeding [ε-¹⁵N]- or [α-¹⁵N]-L-lysine. Our results not only provide information insight into the biosynthesis of anabasine but also suggest an alternative route for the production of anabasine by genetic engineering.

Journal

• Plant Biotechnology

  Plant Biotechnology 31 (5), 511-518, 2014

  Japanese Society for Plant Biotechnology

Keywords

• Anabasine
• tobacco alkaloids
• Lys-derived alkaloids
• lysine/ornithine decarboxylase

Details

• CRID: 1390001204326850560
• NII Article ID: 130004721174
• NII Book ID: AA11250821
• DOI: 10.5511/plantbiotechnology.14.1008a
• ISSN: 13476114
• ISSN: 13424580
• NDL BIB ID: 026092115
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I026092115
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed