Citation: Xincheng Sun, Yuxin Wang, Changkang Li, Ridao Chen, Kebo Xie, Jimei Liu, Songyang Sui, Yaotian Han, Dawei Chen, Jungui Dai. Functional characterization of a glycoside hydrolase in the biosynthesis of camptothecin from Camptotheca acuminata[J]. Chinese Chemical Letters, ;2025, 36(12): 110895. doi: 10.1016/j.cclet.2025.110895 shu

Functional characterization of a glycoside hydrolase in the biosynthesis of camptothecin from Camptotheca acuminata

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State Key Laboratory of Bioactive Substance and Function of Natural Medicines; CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs; and NHC Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China

chendawei@imm.ac.cn

jgdai@imm.ac.cn

Received Date: 21 November 2024
Revised Date: 16 January 2025
Accepted Date: 20 January 2025
Available Online: 21 January 2025

Figures(4)

Camptothecin, a plant-derived pentacyclic pyrroloquinoline alkaloid, and its derivatives like topotecan and irinotecan have been used as clinical anticancer agents for decades. However, the complete biosynthetic pathway of camptothecin still remains unelucidated due to the unknown complex formation processes and corresponding enzymes for the downstream biosynthetic pathway including the committed hydrolysis of glycosides. Herein, a novel glycoside hydrolase (CaGH1) responsible for the deglycosylation of biosynthetic glycoside intermediates including both quinoline-type alkaloids pumiloside (1), (3S)-deoxypumiloside (2) and indole-type alkaloid strictosamide (3) has been functionally identified. Moreover, CaGH1 exhibits the highly strict stereoselectivity towards the substrates with 3S configuration. Furthermore, a combined strategy for the discovery of the unknown biosynthetic enzyme by employing activity-guided enzyme verification, transcriptome-based gene mining, biochemical assay in vitro, and structurally characterizing the unstable enzymatic products by derivatization, is reported. These findings not only provide a better understanding of the deglycosylation in camptothecin biosynthesis, also lay the foundation for the complete elucidation of camptothecin biosynthetic pathway and biological production of camptothecin.
- Glycoside hydrolase,
- Deglycosylation,
- Biosynthesis,
- Camptothecin,
- Camptotheca acuminata
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沈阳化工大学材料科学与工程学院沈阳 110142