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COSY catalyses trans–cis isomerization and lactonization in the biosynthesis of coumarins

Nature Plants volume 5, pages 1066–1075 (2019)Cite this article

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Abstract

Coumarins, also known as 1,2-benzopyrones, comprise a large class of secondary metabolites that are ubiquitously found throughout the plant kingdom. In many plant species, coumarins are particularly important for iron acquisition and plant defence. Here, we show that COUMARIN SYNTHASE (COSY) is a key enzyme in the biosynthesis of coumarins. Arabidopsis thaliana cosy mutants have strongly reduced levels of coumarin and accumulate o-hydroxyphenylpropanoids instead. Accordingly, cosy mutants have reduced iron content and show growth defects when grown under conditions in which there is a limited availability of iron. Recombinant COSY is able to produce umbelliferone, esculetin and scopoletin from their respective o-hydroxycinnamoyl-CoA thioesters by two reaction steps—a trans–cis isomerization followed by a lactonization. This conversion happens partially spontaneously and is catalysed by light, which explains why the need for an enzyme for this conversion has been overlooked. The combined results show that COSY has an essential function in the biosynthesis of coumarins in organs that are shielded from light, such as roots. These findings provide routes to improving coumarin production in crops or by microbial fermentation.

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Fig. 1: Revised coumarin biosynthetic pathway in Arabidopsis displaying metabolic shifts in the cosy mutants.
Fig. 2: Enzymatic activity of COSY.
Fig. 3: cosy mutants are sensitive to media or soil with reduced iron bioavailability.

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Data availability

The raw chromatograms of the phenolic profiling experiment of cosy-1, cosy-2 and wild-type roots are available from the MetaboLights database repository37 as study MTBLS692. The data that support the findings of this study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank A. Bleys for help with preparing the manuscript. We acknowledge funding through the framework of the IWT-SBO (project ARBOREF), the Hercules program of Ghent University for the Synapt Q-Tof (grant no. AUGE/014), the Bijzonder Onderzoeksfonds-Zware Apparatuur of Ghent University for the FT-ICR-MS instrument (174PZA05) and the Multidisciplinary Research Partnership Biotechnology for a Sustainable Economy (01MRB510W) of Ghent University. R.V. was supported by the Research Foundation—Flanders with a postdoctoral fellowship; L.S., L.H. and B.D.M. by the Agency for Innovation by Science and Technology, Flanders (IWT-Vlaanderen); and X.L. and J.L. by the China Scholarship Council. H.K. and J.R. were funded by the DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science DE-SC0018409) and research in the Schmidt laboratory was supported by MoST.

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Author notes

  1. These authors contributed equally: Ruben Vanholme, Lisa Sundin.

Authors and Affiliations

  1. Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, Belgium

    Ruben Vanholme, Lisa Sundin, Keletso Carol Seetso, Xinyu Liu, Jin Li, Barbara De Meester, Lennart Hoengenaert, Geert Goeminne, Kris Morreel, Bartel Vanholme & Wout Boerjan

  2. Center for Plant Systems Biology, VIB, Ghent, Belgium

    Ruben Vanholme, Lisa Sundin, Keletso Carol Seetso, Xinyu Liu, Jin Li, Barbara De Meester, Lennart Hoengenaert, Geert Goeminne, Kris Morreel, Bartel Vanholme & Wout Boerjan

  3. Department of Biochemistry and the DOE Great Lakes Bioenergy Research Center, the Wisconsin Energy Institute, University of Wisconsin, Madison, WI, USA

    Hoon Kim & John Ralph

  4. Metabolomics Core, VIB, Ghent, Belgium

    Geert Goeminne

  5. Protein Core, VIB-UGent Center for Inflammation Research, VIB, Ghent University, Ghent, Belgium

    Jurgen Haustraete

  6. Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan

    Huei-Hsuan Tsai & Wolfgang Schmidt

  7. Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, National Chung Hsing University and Academia Sinica, Taipei, Taiwan

    Huei-Hsuan Tsai & Wolfgang Schmidt

  8. Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan

    Huei-Hsuan Tsai

  9. Biotechnology Center, National Chung Hsing University, Taichung, Taiwan

    Wolfgang Schmidt

  10. Genome and Systems Biology Degree Program, College of Life Science, National Taiwan University, Taipei, Taiwan

    Wolfgang Schmidt

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Contributions

R.V., L.S., W.S. and W.B. designed the experiments. R.V., L.S., K.C.S., H.K., X.L., B.D.M., L.H., J.L., G.G., H.-H.T. and K.M. performed experiments. R.V., L.S., G.G., K.M., J.H., B.V., H.-H.T. and J.R. collected and analysed data; R.V. and W.B. wrote the manuscript with the help of all of the authors.

Corresponding author

Correspondence to Wout Boerjan.

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Competing interests

A patent (Belgium Patent publication no: WO 2018/046526 Al) for the use of COSY for the production of coumarins in plants and microorganisms has been published.

Additional information

Peer review information: Nature Plants thanks Chang Jun Liu and Jing Ke Weng and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–23, legend of Supplementary Table 1, Supplementary Tables 2 and 3, Supplementary Table 4 legend, Supplementary Tables 5 and 6.

Reporting Summary

Supplementary Tables 1 and 4

Supplementary Table 1: a full list of peaks detected in the root extracts of cosy-1 and cosy-2. Supplementary Table 4: a full list of peaks detected in the inflorescence stem extracts of cosy-1 and cosy-2.

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Vanholme, R., Sundin, L., Seetso, K.C. et al. COSY catalyses trans–cis isomerization and lactonization in the biosynthesis of coumarins. Nat. Plants 5, 1066–1075 (2019). https://doi.org/10.1038/s41477-019-0510-0

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