TY - JOUR
T1 - Metabolic diversification of nitrogen-containing metabolites by the expression of a heterologous lysine decarboxylase gene in Arabidopsis
AU - Shimizu, Yohei
AU - Rai, Amit
AU - Okawa, Yuko
AU - Tomatsu, Hajime
AU - Sato, Masaru
AU - Kera, Kota
AU - Suzuki, Hideyuki
AU - Saito, Kazuki
AU - Yamazaki, Mami
N1 - Funding Information:
This work was supported by the JSPS KAKENHI program (grant number 15H02494 to K.S. and 16H06454 to M.Y.), by the Strategic International Collaborative Research Program of Japan Science and Technology Agency (Metabolomics for a Low Carbon Society, JST-NSF) and by the Strategic Priority Research Promotion Program of Chiba University. We thank Dr Hiroshi Tsugawa (RIKEN CSRS, Japan) for assisting with the use of ms-finder and the Arabidopsis transformation service of RIKEN CSRS, Japan. We thank RIKEN BRC for providing cDNA clones. We also thank Dr Megha Rai (Chiba University, Japan), Dr Gourvendu Saxena (National University of Singapore) and Mr Koki Hayashi (Chiba University, Japan) for helping us to improve the text.
Publisher Copyright:
© 2019 The Authors The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd
PY - 2019/11/1
Y1 - 2019/11/1
N2 - Lysine decarboxylase converts l-lysine to cadaverine as a branching point for the biosynthesis of plant Lys-derived alkaloids. Although cadaverine contributes towards the biosynthesis of Lys-derived alkaloids, its catabolism, including metabolic intermediates and the enzymes involved, is not known. Here, we generated transgenic Arabidopsis lines by expressing an exogenous lysine/ornithine decarboxylase gene from Lupinus angustifolius (La-L/ODC) and identified cadaverine-derived metabolites as the products of the emerged biosynthetic pathway. Through untargeted metabolic profiling, we observed the upregulation of polyamine metabolism, phenylpropanoid biosynthesis and the biosynthesis of several Lys-derived alkaloids in the transgenic lines. Moreover, we found several cadaverine-derived metabolites specifically detected in the transgenic lines compared with the non-transformed control. Among these, three specific metabolites were identified and confirmed as 5-aminopentanal, 5-aminopentanoate and δ-valerolactam. Cadaverine catabolism in a representative transgenic line (DC29) was traced by feeding stable isotope-labeled [α-15N]- or [ε-15N]-l-lysine. Our results show similar 15N incorporation ratios from both isotopomers for the specific metabolite features identified, indicating that these metabolites were synthesized via the symmetric structure of cadaverine. We propose biosynthetic pathways for the metabolites on the basis of metabolite chemistry and enzymes known or identified through catalyzing specific biochemical reactions in this study. Our study shows that this pool of enzymes with promiscuous activities is the driving force for metabolite diversification in plants. Thus, this study not only provides valuable information for understanding the catabolic mechanism of cadaverine but also demonstrates that cadaverine accumulation is one of the factors to expand plant chemodiversity, which may lead to the emergence of Lys-derived alkaloid biosynthesis.
AB - Lysine decarboxylase converts l-lysine to cadaverine as a branching point for the biosynthesis of plant Lys-derived alkaloids. Although cadaverine contributes towards the biosynthesis of Lys-derived alkaloids, its catabolism, including metabolic intermediates and the enzymes involved, is not known. Here, we generated transgenic Arabidopsis lines by expressing an exogenous lysine/ornithine decarboxylase gene from Lupinus angustifolius (La-L/ODC) and identified cadaverine-derived metabolites as the products of the emerged biosynthetic pathway. Through untargeted metabolic profiling, we observed the upregulation of polyamine metabolism, phenylpropanoid biosynthesis and the biosynthesis of several Lys-derived alkaloids in the transgenic lines. Moreover, we found several cadaverine-derived metabolites specifically detected in the transgenic lines compared with the non-transformed control. Among these, three specific metabolites were identified and confirmed as 5-aminopentanal, 5-aminopentanoate and δ-valerolactam. Cadaverine catabolism in a representative transgenic line (DC29) was traced by feeding stable isotope-labeled [α-15N]- or [ε-15N]-l-lysine. Our results show similar 15N incorporation ratios from both isotopomers for the specific metabolite features identified, indicating that these metabolites were synthesized via the symmetric structure of cadaverine. We propose biosynthetic pathways for the metabolites on the basis of metabolite chemistry and enzymes known or identified through catalyzing specific biochemical reactions in this study. Our study shows that this pool of enzymes with promiscuous activities is the driving force for metabolite diversification in plants. Thus, this study not only provides valuable information for understanding the catabolic mechanism of cadaverine but also demonstrates that cadaverine accumulation is one of the factors to expand plant chemodiversity, which may lead to the emergence of Lys-derived alkaloid biosynthesis.
KW - Arabidopsis thaliana
KW - Lys-derived alkaloids
KW - cadaverine catabolism
KW - chemodiversity
KW - lysine decarboxylase
KW - non-targeted metabolome analysis
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U2 - 10.1111/tpj.14454
DO - 10.1111/tpj.14454
M3 - Article
C2 - 31364191
AN - SCOPUS:85071175126
VL - 100
SP - 505
EP - 521
JO - Plant Journal
JF - Plant Journal
SN - 0960-7412
IS - 3
ER -