Functional characterization of (S)-N-methylcoclaurine 3'-hydroxylase (NMCH) involved in the biosynthesis of benzylisoquinoline alkaloids in Corydalis yanhusuo

  • Plant Physiol Biochem. 2021 Nov:168:507-515. doi: 10.1016/j.plaphy.2021.09.042.
Xiuyu Liu  1 ,  Junling Bu  2 ,  Ying Ma  3 ,  Yun Chen  4 ,  Qishuang Li  5 ,  Xiang Jiao  6 ,  Zhimin Hu  7 ,  Guanghong Cui  8 ,  Jinfu Tang  9 ,  Juan Guo  10 ,  Luqi Huang  11
Affiliations
  • 1. School of Pharmaceutical Sciences, Henan University of Chinese Medicine, No. 156 Jinshuidong Road, Zhengzhou, 450008, China; State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 2. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 3. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 4. Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE41296, Gothenburg, Sweden. Electronic address: [email protected].
  • 5. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 6. Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE41296, Gothenburg, Sweden. Electronic address: [email protected].
  • 7. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 8. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 9. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 10. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
  • 11. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, NO.16 Neinanxiaojie, Dongcheng district, Beijing, China. Electronic address: [email protected].
PMID: 34757301 DOI: 10.1016/j.plaphy.2021.09.042
Abstract

Benzylisoquinoline Alkaloids (BIAs) are compounds naturally found in Plants and can have significant value in clinical settings. Metabolic engineering and synthetic biology are both promising approaches for the heterologous acquisition of benzylisoquinoline Alkaloids. (S)-N-methylcoclaurine 3'-hydroxylase (NMCH), a member of the CYP80 family of CYP450, is the penultimate catalytic enzyme that forms the central branch-point intermediate (S)-reticuline and plays a key role in the biosynthesis of BIAs. In this study, an NMCH gene was cloned from Corydalis yanhusuo, while in vitro reactions demonstrated that CyNMCH can catalyze (S)-N-methylcoclaurine to produce (S)-3'-hydroxy-N-methylcoclaurine. The Km and Kcat of CyNMCH were estimated and compared with those identified in Eschscholzia californica and Coptis japonica. This newly discovered CyNMCH will provide alternative genetic resources for the synthetic biological production of benzylisoquinoline Alkaloids and provides a foundation to help analyze the biosynthetic pathway of BIAs biosynthesis in C. yanhusuo.

Keywords
(S)–N-Methylcoclaurine 3′-hydroxylase; Corydalis yanhusuo; Synthetic biology.
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