Presentation Information
[P04-513]Development of Culture-Based Production Systems for the Sustainable Production of Phenylethanoid Glycosides
○Yushiro Fuji1,2, Hiroshi Matsufuji2, Masami Yokota Hirai1,3 (1. RIKEN Center for Sustainable Resource Science (Japan), 2. Department of Food Science and Technology, College of Bioresource Sciences, Nihon University (Japan), 3. Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University (Japan))
Keywords:
Phenylethanoid glycoside,Biosynthesis pathway,Culture Production
[Purpose] Phenylethanoid glycosides (PhGs) are specialized metabolites characterized by a structure in which a C6-C2 phenylethanoid moiety is linked to a glucoside core, and they are widely distributed in medicinal and aromatic plants. Among them, acteoside is a representative PhG that has attracted attention because of its diverse biological and pharmacological activities, including antioxidant, anti-inflammatory, and neuroprotective effects. However, sustainable production systems for acteoside and related PhGs have not yet been established, mainly because their biosynthetic pathways remain insufficiently understood and efficient biological production systems are lacking. In our previous studies, we found that sesame (Sesamum indicum L.) accumulates acteoside at exceptionally high levels in its leaf blades, reaching up to 12.3% of dry weight, one of the highest levels reported in plants. Therefore, this study aimed to elucidate the biosynthetic pathway of acteoside and to develop culture-based production systems using sesame.
[Method] Calli were induced from sesame hypocotyls, and a suspension-cultured cell system was subsequently established. We also examined elicitor-induced acteoside production using various treatments and performed transcriptome analysis of the treated cells. Based on the resulting expression profiles, genes encoding glycosyltransferases and acyltransferases involved in the downstream steps of the biosynthetic process were selected as candidate genes. Furthermore, we established a sesame hairy root culture system as a functional platform for analyzing candidate regulatory genes. Candidate transcription factors were selected from transcriptome datasets derived from cultured cells, and their effects were evaluated by overexpressing them in the sesame hairy root culture system.
[Results] Methyl jasmonate elicitation of the suspension-cultured cells increased acteoside content 6.6-fold after 72 h. Enzyme assays in Escherichia coli expressing candidate genes selected from transcriptome-based expression profiles identified glycosyltransferases and acyltransferases involved in the downstream steps of the biosynthetic pathway. Furthermore, acteoside accumulation was confirmed in the established hairy root system, and overexpression analysis suggested that at least some candidate regulatory factors promote acteoside accumulation in sesame hairy roots.
[Consideration] These results suggest that sesame is a useful system for studying phenylethanoid glycoside biosynthesis. The identified downstream enzymes and their substrate specificities provided insight into the likely reaction sequence in sesame, including the order of glycosylation and acylation steps. The hairy root system also offers a practical tool for functional analysis of candidate regulatory genes. Together, these findings indicate that sesame can serve as both a natural high producer of acteoside and an experimental system for developing culture-based production methods.
[Conclusion] Sesame is a promising model for elucidating PhG biosynthesis and developing sustainable production strategies. Our findings provide a basis for future metabolic engineering and regulatory studies aimed at the stable production of valuable PhGs.
[Method] Calli were induced from sesame hypocotyls, and a suspension-cultured cell system was subsequently established. We also examined elicitor-induced acteoside production using various treatments and performed transcriptome analysis of the treated cells. Based on the resulting expression profiles, genes encoding glycosyltransferases and acyltransferases involved in the downstream steps of the biosynthetic process were selected as candidate genes. Furthermore, we established a sesame hairy root culture system as a functional platform for analyzing candidate regulatory genes. Candidate transcription factors were selected from transcriptome datasets derived from cultured cells, and their effects were evaluated by overexpressing them in the sesame hairy root culture system.
[Results] Methyl jasmonate elicitation of the suspension-cultured cells increased acteoside content 6.6-fold after 72 h. Enzyme assays in Escherichia coli expressing candidate genes selected from transcriptome-based expression profiles identified glycosyltransferases and acyltransferases involved in the downstream steps of the biosynthetic pathway. Furthermore, acteoside accumulation was confirmed in the established hairy root system, and overexpression analysis suggested that at least some candidate regulatory factors promote acteoside accumulation in sesame hairy roots.
[Consideration] These results suggest that sesame is a useful system for studying phenylethanoid glycoside biosynthesis. The identified downstream enzymes and their substrate specificities provided insight into the likely reaction sequence in sesame, including the order of glycosylation and acylation steps. The hairy root system also offers a practical tool for functional analysis of candidate regulatory genes. Together, these findings indicate that sesame can serve as both a natural high producer of acteoside and an experimental system for developing culture-based production methods.
[Conclusion] Sesame is a promising model for elucidating PhG biosynthesis and developing sustainable production strategies. Our findings provide a basis for future metabolic engineering and regulatory studies aimed at the stable production of valuable PhGs.
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