Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression

doi:10.1007/s13659-026-00601-7

Natural Products and Bioprospecting ›› 2026, Vol. 16 ›› Issue (4): 49-49.DOI: 10.1007/s13659-026-00601-7

• ORIGINAL ARTICLES • Previous Articles Next Articles

Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression

Ziou Zha¹, Dan He¹, Jianguo Song², Zhenhua Guan³, Jiapei Han¹, Chang Liu¹, Xinyu Wang¹, Yongchun Zhu¹, Hucheng Zhu¹, Wencai Ye², Qin Li¹, Yonghui Zhang¹, Yuan Zhou¹

1. Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, People's Republic of China;
2. State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China;
3. Department of Pharmacy, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital), Southern University of Science and Technology, Shenzhen 518055, Guangdong, People's Republic of China

Received:2026-01-06 Accepted:2026-02-03 Online:2026-08-29 Published:2026-08-22
Contact: Qin Li,E-mail:liqin2023@hust.edu.cn;Yonghui Zhang,E-mail:zhangyh@mails.tjmu.edu.cn;Yuan Zhou,E-mail:zhouyuan@hust.edu.cn
Supported by:
This work was financially supported by the National Key R&D Program of China (No. 2021YFA0910500), the National Natural Science Foundation of China (No. 81973205, 22477036, 22277035, and U22A20380), the Fundamental Research Funds for the Central Universities (2024BRA018), and the Sino-German Center for Research Promotion (M-0477).

Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression

Ziou Zha¹, Dan He¹, Jianguo Song², Zhenhua Guan³, Jiapei Han¹, Chang Liu¹, Xinyu Wang¹, Yongchun Zhu¹, Hucheng Zhu¹, Wencai Ye², Qin Li¹, Yonghui Zhang¹, Yuan Zhou¹

1. Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, People's Republic of China;
2. State Key Laboratory of Bioactive Molecules and Druggability Assessment, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China;
3. Department of Pharmacy, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital), Southern University of Science and Technology, Shenzhen 518055, Guangdong, People's Republic of China

通讯作者: Qin Li,E-mail:liqin2023@hust.edu.cn;Yonghui Zhang,E-mail:zhangyh@mails.tjmu.edu.cn;Yuan Zhou,E-mail:zhouyuan@hust.edu.cn
基金资助:
This work was financially supported by the National Key R&D Program of China (No. 2021YFA0910500), the National Natural Science Foundation of China (No. 81973205, 22477036, 22277035, and U22A20380), the Fundamental Research Funds for the Central Universities (2024BRA018), and the Sino-German Center for Research Promotion (M-0477).

Abstract

Abstract: Prenylation modifications of natural products typically introduce greater structural complexity and enhance their biological activities. Yet, the modification of piperazine alkaloids by dimethylallyl tryptophan synthases (DMATS) remains unreported. In this study, we identified and activated a silent DMATS-containing piperazine biosynthetic gene cluster (BGC), flz, in Aspergillus flavipes via heterologous expression and in vitro enzymatic assay, leading to the isolation and identification of sixteen metabolites. Among these, twelve are new, including five tryptophan-valine-derived alkaloids (2-6) and seven previously unreported prenylated analogs (8-13, 17). Notably, 8 and 9 represent novel prenylated piperazines featuring a unique 6-5-5-6 ring system. Most significantly, we uncovered a versatile DMATS, FlzE, capable of catalyzing mono-prenylation on flexible substrates, such as piperazine, pyrazine, and diketopiperazine, at multiple sites in either regular or reverse manners. This study not only expands the chemical space of indole alkaloid derivatives but also provides a versatile and engineerable biocatalyst for the prenylation of both natural and synthetic products.

Key words: Aspergillus flavipes, Genome mining, Biosynthesis, Piperazine alkaloids, Dimethylallyl tryptophan synthases

摘要： Prenylation modifications of natural products typically introduce greater structural complexity and enhance their biological activities. Yet, the modification of piperazine alkaloids by dimethylallyl tryptophan synthases (DMATS) remains unreported. In this study, we identified and activated a silent DMATS-containing piperazine biosynthetic gene cluster (BGC), flz, in Aspergillus flavipes via heterologous expression and in vitro enzymatic assay, leading to the isolation and identification of sixteen metabolites. Among these, twelve are new, including five tryptophan-valine-derived alkaloids (2-6) and seven previously unreported prenylated analogs (8-13, 17). Notably, 8 and 9 represent novel prenylated piperazines featuring a unique 6-5-5-6 ring system. Most significantly, we uncovered a versatile DMATS, FlzE, capable of catalyzing mono-prenylation on flexible substrates, such as piperazine, pyrazine, and diketopiperazine, at multiple sites in either regular or reverse manners. This study not only expands the chemical space of indole alkaloid derivatives but also provides a versatile and engineerable biocatalyst for the prenylation of both natural and synthetic products.

关键词: Aspergillus flavipes, Genome mining, Biosynthesis, Piperazine alkaloids, Dimethylallyl tryptophan synthases

Ziou Zha, Dan He, Jianguo Song, Zhenhua Guan, Jiapei Han, Chang Liu, Xinyu Wang, Yongchun Zhu, Hucheng Zhu, Wencai Ye, Qin Li, Yonghui Zhang, Yuan Zhou. Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression[J]. Natural Products and Bioprospecting, 2026, 16(4): 49-49.

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Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression

Discovery of unprecedented prenylated indole piperazines and pyrazines through cryptic biosynthetic gene cluster heterologous expression

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