Abstract
Although Z-2-oxo-4-methyl-3-pentene-1,5-dioic acid (Z-OMPD) has been identified as a major dicarboxylic acid in tulip tissues, its biosynthetic pathway has not been elucidated. Herein, Z-OMPD was isolated from tulip leaves and chemically synthesized. Comparisons of these samples revealed that Z-OMPD exists as a tautomeric mixture at physiological pH. As a regioisomer of Z-OMPD, we enzymatically and chemically prepared 4-methylene-2-oxo-glutaric acid (4-MEOG) for the first time. Using these compounds as standards, the occurrence of Z-OMPD and 4-MEOG in various tissues of the tulip cultivar “Murasakizuisho” was evaluated directly and by 2,4-dinitrophenylhydrazone derivatization. Z-OMPD was found to be abundant in the aerial tissues, whereas 4-MEOG was almost absent from all tissues. Stability analyses of Z-OMPD and 4-MEOG revealed that no double bond isomerization occurred at physiological pH, suggesting that enzyme systems are responsible for Z-OMPD biosynthesis in tulip tissues.
Funding source: Japan Society for the Promotion of Science
Award Identifier / Grant number: 18K05463
Award Identifier / Grant number: 21K05403
Acknowledgements
The authors thank Dr. Issei Osaka, Toyama Prefectural University, for high-resolution mass spectrometry measurements.
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Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
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Research funding: This work was supported in part by Japan Society for the Promotion of Science KAKENHI grant nos. 18K05463 (to TN) and 21K05403 (to YK).
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Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
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Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/znc-2021-0282).
S1: General experimental details, S2.1: isolation of 4-MnGlu, S2.2: isolation of MnGln, S2.3–S2.8; synthesis of chemical compounds, S3: enzymatic deamination of 4-MnGlu by GDH, S4: expression and purification of recombinant GOx, S5: 2,4-DH derivatization of the crude extract of tulip leaves, and Figures S1–S5 are available in the Supplementary Material.
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