Systematic identification of terpene synthases from sacred lotus (Nelumbo nucifera) and heterologous biosynthesis of the insecticidal and antimicrobial compound γ-eudesmol

Abstract

Sacred lotus is widely used in the agricultural, nutraceutical, and pharmaceutical industries. Terpenes are not only crucial components of sacred lotus essential oil, but also serve as signaling molecules involved in plant-environment interactions. However, the biosynthesis of terpenes in sacred lotus has not yet been reported. Thus, gene-directed heterologous mining and combinatorial biosynthesis methods were used in this study to systematically characterize the function of terpene synthase genes in the sacred lotus. As a result, two monoterpene, 11 sesquiterpene, and three diterpene products were synthesized, and a highly efficient γ-eudesmol synthase was discovered. In addition, a mechanistic study revealed that N314 is the key amino acid responsible for the secondary cyclization that produces γ-eudesmol. In vitro assays demonstrated that γ-eudesmol exhibited substantial insecticidal and antimicrobial activities. Furthermore, de novo biosynthesis of γ-eudesmol was achieved in a yeast chassis through a series of metabolic engineering strategies, reaching a titer of 801.66 mg/L in a shake flask, the highest yield reported to date. The present study uncovered the biosynthesis of terpenes in sacred lotus, as well as successfully synthesized the bioactive compound γ-eudesmol by synthetic biology. This comprehensive strategy can be readily adapted for investigation and the production of other valuable plant-derived natural products.

Acknowledgments

We gratefully acknowledge Prof. Xianbao Deng from Wuhan Botanical Garden for kindly providing the sacred lotus (Nelumbo nucifera) plant materials used in this study. We sincerely thank Dr. Wu Meiling for her thorough linguistic revision of the manuscript. The numerical calculations in this paper have been done on the supercomputing system in the Supercomputing Center of Wuhan University. This work is supported by grants from the National Key Research and Development Program (2022YFA0912100).

Author contributions

Z.X., X.F., Y.Z., X.X., J.Y., J.H., H.Z., F.C., and W.C. performed the experiments. Z.X., X.F., and L.L. conceived the project, designed the experiments, and prepared the manuscript. T.L. discussed the results and contributed to the final manuscript. All authors read and approved the final manuscript.

Data availability

Sequences of NnTPSs in this study were provided in Table S2.

Conflict of interest statement:

The authors have applied for a patent based on this work.

Supplementary data

Supplementary data is available at Horticulture Research online.

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