Identification of tuliposides K–M in tulip bulbs via an enzyme reaction-based screening method using a tuliposide-converting enzyme

Taiji Nomura; Ayami Omode; Yasuo Kato

doi:10.1515/znc-2023-0068

Abstract

Tuliposides (Pos) are major defense-related secondary metabolites in tulip, having 4-hydroxy-2-methylenebutanoyl and/or (3S)-3,4-dihydroxy-2-methylenebutanoyl groups at the C-1 and/or C-6 positions of d-glucose. The acyl group at the C-6 position is converted to antimicrobial lactones (tulipalins) by an endogenous Pos-converting enzyme. Based on this enzyme activity, we examined tulip bulb extracts and detected HPLC peaks that disappeared following the reaction by the Pos-converting enzyme. Spectroscopic analyses of the three purified compounds revealed that one of them was a glucose ester-type Pos, while the other two were identified as a glucoside ester-type Pos. These compounds were designated as PosK, L, and M. They were specific to bulbs, with the highest content in the outermost layer, but they were markedly less abundant than PosG, the minor bulb Pos we identified earlier. The study results suggest that tulip bulbs contain at least four minor Pos in addition to the major 6-PosA. Although PosK–M were present in almost all of the tested tulip cultivars, they were detected in only a few wild species, indicative of their potential utility as chemotaxonomic markers in tulip. Identification of PosK–M as 6-PosA derivatives unveils the biosynthetic diversity of Pos, the well-known group of secondary metabolites in tulip.

Keywords: glucose ester; glucoside ester; tulip; tulipalin; tuliposide

Corresponding author: Taiji Nomura, Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan, E-mail: tnomura@pu-toyama.ac.jp

Funding source: Japan Society for the Promotion of Science

Award Identifier / Grant number: JP18K05463

Acknowledgments

We thank Ms. Ran Masumoto and Ms. Mika Kamiguchi (Toyama Prefectural University) for technical assistance. This work was supported by JSPS KAKENHI grant no. JP18K05463 (to TN). We thank Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Conflict of interest statement: The authors declare no conflicts of interest regarding this article.

References

1. Tschesche, R, Kämmerer, F-J, Wulff, G, Schönbeck, F. Über die antibiotisch wirksamen substanzen der tulpe (Tulipa gesneriana). Tetrahedron Lett 1968;9:701–6. https://doi.org/10.1016/s0040-4039(00)75615-2.Search in Google Scholar PubMed

2. Tschesche, R, Kämmerer, F-J, Wulff, G. Über die struktur der antibiotisch aktiven substanzen der tulpe (Tulipa gesneriana L.). Chem Ber 1969;102:2057–71. https://doi.org/10.1002/cber.19691020631.Search in Google Scholar

3. Nomura, T. Function and application of a non-ester-hydrolyzing carboxylesterase discovered in tulip. Biosci Biotechnol Biochem 2017;81:81–94. https://doi.org/10.1080/09168451.2016.1240608.Search in Google Scholar PubMed

4. Christensen, LP, Kristiansen, K. Isolation and quantification of tuliposides and tulipalins in tulips (Tulipa) by high-performance liquid chromatography. Contact Dermatitis 1999;40:300–9. https://doi.org/10.1111/j.1600-0536.1999.tb06080.x.Search in Google Scholar PubMed

5. Kato, Y, Shoji, K, Ubukata, M, Shigetomi, K, Sato, Y, Nakajima, N, et al.. Purification and characterization of a tuliposide-converting enzyme from bulbs of Tulipa gesneriana. Biosci Biotechnol Biochem 2009;73:1895–7. https://doi.org/10.1271/bbb.90226.Search in Google Scholar PubMed

6. Kato, Y, Yoshida, H, Shoji, K, Sato, Y, Nakajima, N, Ogita, S. A facile method for the preparation of α-methylene-γ-butyrolactones from tulip tissues by enzyme-mediated conversion. Tetrahedron Lett 2009;50:4751–3. https://doi.org/10.1016/j.tetlet.2009.06.018.Search in Google Scholar

7. Kato, Y, Futanaga, T, Nomura, T. Substrate specificity of tuliposide-converting enzyme, a unique non-ester-hydrolyzing carboxylesterase in tulip: effects of the alcohol moiety of substrate on the enzyme activity. Bioorg Med Chem Lett 2019;29:664–7. https://doi.org/10.1016/j.bmcl.2018.12.010.Search in Google Scholar PubMed

8. Nomura, T, Ogita, S, Kato, Y. A novel lactone-forming carboxylesterase: molecular identification of a tuliposide A-converting enzyme in tulip. Plant Physiol 2012;159:565–78. https://doi.org/10.1104/pp.112.195388.Search in Google Scholar PubMed PubMed Central

9. Nomura, T, Tsuchigami, A, Ogita, S, Kato, Y. Molecular diversity of tuliposide A-converting enzyme in the tulip. Biosci Biotechnol Biochem 2013;77:1042–8. https://doi.org/10.1271/bbb.130021.Search in Google Scholar PubMed

10. Nomura, T, Murase, T, Ogita, S, Kato, Y. Molecular identification of tuliposide B-converting enzyme: a lactone-forming carboxylesterase from the pollen of tulip. Plant J 2015;83:252–62. https://doi.org/10.1111/tpj.12883.Search in Google Scholar PubMed

11. Nomura, T, Ueno, A, Ogita, S, Kato, Y. Molecular diversity of tuliposide B-converting enzyme in tulip (Tulipa gesneriana): identification of the root-specific isozyme. Biosci Biotechnol Biochem 2017;81:1185–93. https://doi.org/10.1080/09168451.2017.1295806.Search in Google Scholar PubMed

12. Nomura, T, Kuchida, R, Kitaoka, N, Kato, Y. Molecular diversity of tuliposide B-converting enzyme in tulip (Tulipa gesneriana): identification of the third isozyme with a distinct expression profile. Biosci Biotechnol Biochem 2018;82:810–20. https://doi.org/10.1080/09168451.2018.1438170.Search in Google Scholar PubMed

13. Christensen, LP. A further tuliposide from Alstroemeria revoluta. Phytochemistry 1995;40:49–51. https://doi.org/10.1016/0031-9422(95)00235-y.Search in Google Scholar

14. Lee, C-L, Gao, Z-A, Jhan, Y-L, Chang, Y-S, Chen, C-J. Tuliposides H-J and bioactive components from the bulb of Amana edulis. Molecules 2021;26:5907. https://doi.org/10.3390/molecules26195907.Search in Google Scholar PubMed PubMed Central

15. Christensen, LP. Tuliposides from Tulipa sylvestris and T. turkestanica. Phytochemistry 1999;51:969–74. https://doi.org/10.1016/s0031-9422(98)00716-x.Search in Google Scholar

16. Nomura, T, Ogita, S, Kato, Y. Isolation and identification of tuliposides D and F from tulip cultivars. Z Naturforsch C Biosci 2020;75:7–12. https://doi.org/10.1515/znc-2019-0123.Search in Google Scholar PubMed

17. Nomura, T, Ogita, S, Kato, Y. One-step enzymatic synthesis of 1-tuliposide A using tuliposide-converting enzyme. Appl Biochem Biotechnol 2019;188:12–28. https://doi.org/10.1007/s12010-018-2903-3.Search in Google Scholar PubMed

18. Nomura, T, Kato, Y. Identification of tuliposide G, a novel glucoside ester-type tuliposide, and its distribution in tulip. Z Naturforsch C Biosci 2020;75:75–86. https://doi.org/10.1515/znc-2019-0176.Search in Google Scholar PubMed

19. Lepore, L, Malafronte, N, Condero, FB, Gualtieri, MJ, Abdo, S, Piaz, FD, et al.. Isolation and structural characterization of glycosides from an anti-angiogenic extract of Monnina obtusifolia H.B.K. Fitoterapia 2011;82:178–83. https://doi.org/10.1016/j.fitote.2010.08.018.Search in Google Scholar PubMed

20. Christensen, LP, Kristiansen, K. Isolation and quantification of a new tuliposide (tuliposide D) by HPLC in Alstroemeria. Contact Dermatitis 1995;33:188–92. https://doi.org/10.1111/j.1600-0536.1995.tb00543.x.Search in Google Scholar PubMed

21. Christensen, LP. Tuliposides from Alstroemeria revoluta. Phytochemistry 1995;38:1371–3. https://doi.org/10.1016/0031-9422(94)00809-8.Search in Google Scholar

22. Binutu, OA, Cordell, GA. Constituents of Afzelia bella stem bark. Phytochemistry 2001;56:827–30. https://doi.org/10.1016/s0031-9422(01)00006-1.Search in Google Scholar PubMed

23. Perry, NB, Brennan, NJ. Antimicrobial and cytotoxic phenolic glycoside esters from the New Zealand tree Toronia toru. J Nat Prod 1997;60:623–6. https://doi.org/10.1021/np970013z.Search in Google Scholar PubMed

24. MacLeod, JK, Rasmussen, HB, Willis, AC. A new glycoside antimicrobial agent from Persoonia linearis × pinifolia. J Nat Prod 1997;60:620–2. https://doi.org/10.1021/np970006a.Search in Google Scholar PubMed

25. Yoshikawa, K, Kinoshita, H, Kan, Y, Arihara, S. Neolignans and phenylpropanoids from the rhizomes of Coptis japonica var. dissecta. Chem Pharm Bull 1995;43:578–81. https://doi.org/10.1248/cpb.43.578.Search in Google Scholar

26. Yoshikawa, K, Kinoshita, H, Arihara, S. Non-basic components of Coptis rhizoma. II. Four new hemiterpenoid glucosides, two new phenylpropanoid glucosides and a new flavonoid glycoside from Coptis japonica var. dissecta. Nat Med 1997;51:244–8.Search in Google Scholar

27. Hiradate, S, Morita, S, Sugie, H, Fujii, Y, Harada, J. Phytotoxic cis-cinnamoyl glucosides from Spiraea thunbergii. Phytochemistry 2004;65:731–9. https://doi.org/10.1016/j.phytochem.2004.01.010.Search in Google Scholar PubMed

28. Park, SH, Park, KH, Oh, MH, Kim, HH, Choe, KI, Kim, SR, et al.. Anti-oxidative and anti-inflammatory activities of caffeoyl hemiterpene glycosides from Spiraea prunifolia. Phytochemistry 2013;96:430–6. https://doi.org/10.1016/j.phytochem.2013.09.017.Search in Google Scholar PubMed

29. Christenhusz, MJM, Govaerts, R, David, JC, Hall, T, Boland, K, Roberts, PS, et al.. Tiptoe through the tulips – cultural history, molecular phylogenetics and classification of Tulipa (Liliaceae). Bot J Linn Soc 2013;172:280–328. https://doi.org/10.1111/boj.12061.Search in Google Scholar

30. Marasek-Ciolakowska, A, Ramanna, MS, Arens, P, Van Tuyl, JM. Breeding and cytogenetics in the genus Tulipa. Floric Ornam Biotechnol 2012;6:90–7.Search in Google Scholar

31. Wilford, R. Tulips: species and hybrids for the gardener. Portland: Timber Press; 2006.Search in Google Scholar

32. Nomura, T, Hayashi, E, Kawakami, S, Ogita, S, Kato, Y. Environmentally benign process for the preparation of antimicrobial α-methylene-β-hydroxy-γ-butyrolactone (tulipalin B) from tulip biomass. Biosci Biotechnol Biochem 2015;79:25–35. https://doi.org/10.1080/09168451.2014.946395.Search in Google Scholar PubMed

Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/znc-2023-0068).

Received: 2023-05-05

Accepted: 2023-06-21

Published Online: 2023-07-05

Published in Print: 2023-09-26

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Supplementary Material