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Characterization of CYP82 genes involved in the biosynthesis of structurally diverse benzylisoquinoline alkaloids in Corydalis yanhusuo

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Abstract

Benzylisoquinoline alkaloids (BIAs) represent a significant class of secondary metabolites with crucial roles in plant physiology and substantial potential for clinical applications. CYP82 genes are involved in the formation and modification of various BIA skeletons, contributing to the structural diversity of compounds. In this study, Corydalis yanhusuo, a traditional Chinese medicine rich in BIAs, was investigated to identify the catalytic function of CYP82s during BIA formation. Specifically, 20 CyCYP82-encoding genes were cloned, and their functions were identified in vitro. Ten of these CyCYP82s were observed to catalyze hydroxylation, leading to the formation of protopine and benzophenanthridine scaffolds. Furthermore, the correlation between BIA accumulation and the expression of CyCYP82s in different tissues of C. yanhusuo was assessed their. The identification and characterization of CyCYP82s provide novel genetic elements that can advance the synthetic biology of BIA compounds such as protopine and benzophenanthridine, and offer insights into the biosynthesis of BIAs with diverse structures in C. yanhusuo.

Key message

CYP82s identified from Corydalis yanhusuo can catalyze the hydroxylation of different substrates. These newly identified CYP82s provide genetic elements for the production of BIAs by synthetic biology.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (82011530137 and 82304650), Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences (CI2021B014), Key project at central government level: The ability to establish sustainable use of valuable Chinese medicine resources (2060302), and the Fundamental Research Funds of China Academy of Chinese Medical Sciences (ZZ14-YQ-044 and ZZ13-YQ-083), Henan Province Chinese herbal medicine industry technology system (Henan Agricultural Science and Education [2018] No.14), The special subsidy project for public health services of traditional Chinese medicine “National Survey of Traditional Chinese Medicine Resources” (Yucai [2019] No.40).

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Henan University of Chinese Medicine, No. 156 Jinshuidong Road, Zhengzhou, 450046, China

    Xiuyu Liu, Conglong Lian, Rui Ma & Suiqing Chen

  2. State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, Chinan Academy of Chinese Medical Sciences, Beijing, 100000, China

    Ying Ma, Junling Bu, Qishuang Li, Zhimin Hu, Juan Guo & Luqi Huang

  3. Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, 41296, Gothenburg, Sweden

    Xiang Jiao & Yun Chen

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  1. Xiuyu Liu
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  2. Ying Ma
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  3. Junling Bu
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  10. Suiqing Chen
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  11. Juan Guo
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  12. Luqi Huang
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Contributions

Conceived and designed the experiment: Luqi Huang, Juan Guo. Performed the experiments: Xiuyu Liu, Junling Bu. Ying Ma, Qishuang Li, and Zhimin Hu analyzed the data. Contributed reagents/materials/analysis tools: Xiang Jiao and Yun Chen. Wrote the manuscript: Xiuyu Liu, Juan Guo. Helped in data analysis and manuscript preparation: Suiqing Chen, Conglong Lian, Rui Ma.

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Correspondence to Suiqing Chen, Juan Guo or Luqi Huang.

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Liu, X., Ma, Y., Bu, J. et al. Characterization of CYP82 genes involved in the biosynthesis of structurally diverse benzylisoquinoline alkaloids in Corydalis yanhusuo. Plant Mol Biol 114, 23 (2024). https://doi.org/10.1007/s11103-023-01397-7

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