Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus

doi:10.1007/s13659-025-00578-9

Natural Products and Bioprospecting ›› 2026, Vol. 16 ›› Issue (2): 27-27.DOI: 10.1007/s13659-025-00578-9

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Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus

Ming-Xuan Gao¹, Li-Li Guo¹, Meng-Ting Wang¹, Xin-Yi Zhang¹, Xinyang Li¹, Shin-Yee Fung², He-Ping Chen¹, Ji-Kai Liu¹

1. International Cooperation Base for Active Substances in Traditional Chinese Medicine in Hubei Province, School of Pharmaceutical Sciences, South-Central Minzu University, Wuhan, China;
2. Medicinal Mushroom Research Group (MMRG), Mycological Sciences Laboratory, Department of Molecular Medicine, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia

Received:2025-10-23 Online:2026-04-22 Published:2026-04-22
Contact: He-Ping Chen,Email:chenhp@mail.scuec.edu.cn;Ji-Kai Liu,Email:liujikai@mail.scuec.edu.cn
Supported by:
National Natural Science Foundation of China, 22177138, Ji-Kai Liu, Fundamental Research Funds for the Central Universities, South-Central Minzu University, PTZ25025, Heping Chen

Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus

Ming-Xuan Gao¹, Li-Li Guo¹, Meng-Ting Wang¹, Xin-Yi Zhang¹, Xinyang Li¹, Shin-Yee Fung², He-Ping Chen¹, Ji-Kai Liu¹

1. International Cooperation Base for Active Substances in Traditional Chinese Medicine in Hubei Province, School of Pharmaceutical Sciences, South-Central Minzu University, Wuhan, China;
2. Medicinal Mushroom Research Group (MMRG), Mycological Sciences Laboratory, Department of Molecular Medicine, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia

通讯作者: He-Ping Chen,Email:chenhp@mail.scuec.edu.cn;Ji-Kai Liu,Email:liujikai@mail.scuec.edu.cn
基金资助:
National Natural Science Foundation of China, 22177138, Ji-Kai Liu, Fundamental Research Funds for the Central Universities, South-Central Minzu University, PTZ25025, Heping Chen

Abstract

Abstract: The Tiger Milk mushrooms (Lignosus spp.) have long been used as traditional folk medicines throughout Southeast Asia. However, the chemical constituents of these species remain largely unexplored. In this study, four type I sesquiterpene synthases (LrhTS1-LrhTS4) from Lignosus rhinocerus TM02^® were functionally characterized through genome mining, in vitro enzymatic assays, and heterologous expression in an Escherichia coli host engineered to overexpress an exogenous mevalonate pathway. The enzyme products were analyzed by GC-MS and further purified and structurally elucidated by NMR spectroscopy. A total of 11 sesquiterpenes (1-11) were identified, among which two were unstable and converted to stable derivatives (2 to 2a and 7 to 7a). Notably, compounds 1-10 are reported from the genus Lignosus for the first time. LrhTS1 and LrhTS3 exhibited high catalytic specificity, whereas LrhTS2 and LrhTS4 displayed product promiscuity. This work expands the knowledge of terpene synthase from mushroom, and advances the understanding of the chemical basis underlying the bioactivity of the Lignosus rhinocerus TM02^®.

Key words: Lignosus rhinocerus (Cooke) Ryvarden, Tiger Milk Mushroom, Sesquiterpene synthase, Heterologous expression

摘要： The Tiger Milk mushrooms (Lignosus spp.) have long been used as traditional folk medicines throughout Southeast Asia. However, the chemical constituents of these species remain largely unexplored. In this study, four type I sesquiterpene synthases (LrhTS1-LrhTS4) from Lignosus rhinocerus TM02^® were functionally characterized through genome mining, in vitro enzymatic assays, and heterologous expression in an Escherichia coli host engineered to overexpress an exogenous mevalonate pathway. The enzyme products were analyzed by GC-MS and further purified and structurally elucidated by NMR spectroscopy. A total of 11 sesquiterpenes (1-11) were identified, among which two were unstable and converted to stable derivatives (2 to 2a and 7 to 7a). Notably, compounds 1-10 are reported from the genus Lignosus for the first time. LrhTS1 and LrhTS3 exhibited high catalytic specificity, whereas LrhTS2 and LrhTS4 displayed product promiscuity. This work expands the knowledge of terpene synthase from mushroom, and advances the understanding of the chemical basis underlying the bioactivity of the Lignosus rhinocerus TM02^®.

关键词: Lignosus rhinocerus (Cooke) Ryvarden, Tiger Milk Mushroom, Sesquiterpene synthase, Heterologous expression

Ming-Xuan Gao, Li-Li Guo, Meng-Ting Wang, Xin-Yi Zhang, Xinyang Li, Shin-Yee Fung, He-Ping Chen, Ji-Kai Liu. Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus[J]. Natural Products and Bioprospecting, 2026, 16(2): 27-27.

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Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus

Genome mining and functional characterization of four type I sesquiterpene synthases from the Tiger Milk Mushroom Lignosus rhinocerus

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