Identification and characterization of camptothecin tailoring enzymes in Nothapodytes tomentosa
© 2024 Institute of Botany, Chinese Academy of Sciences..
Camptothecin is a complex monoterpenoid indole alkaloid with remarkable antitumor activity. Given that two C-10 modified camptothecin derivatives, topotecan and irinotecan, have been approved as potent anticancer agents, there is a critical need for methods to access other aromatic ring-functionalized congeners (e.g., C-9, C-10, etc.). However, contemporary methods for chemical oxidation are generally harsh and low-yielding when applied to the camptothecin scaffold, thereby limiting the development of modified derivatives. Reported herein, we have identified four tailoring enzymes responsible for C-9 modifications of camptothecin from Nothapodytes tomentosa, via metabolomic and transcriptomic analysis. These consist of a cytochrome P450 (NtCPT9H) which catalyzes the regioselective oxidation of camptothecin to 9-hydroxycamptothecin, as well as two methyltransferases (NtOMT1/2, converting 9-hydroxycamptothecin to 9-methoxycamptothecin), and a uridine diphosphate-glycosyltransferase (NtUGT5, decorating 9-hydroxycamptothecin to 9-β-D-glucosyloxycamptothecin). Importantly, the critical residues that contribute to the specific catalytic activity of NtCPT9H have been elucidated through molecular docking and mutagenesis experiments. This work provides a genetic basis for producing camptothecin derivatives through metabolic engineering. This will hasten the discovery of novel C-9 modified camptothecin derivatives, with profound implications for pharmaceutical manufacture.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - year:2024 |
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| Enthalten in: |
Journal of integrative plant biology - (2024) vom: 22. März |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Chen, Yin [VerfasserIn] |
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| Links: |
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| Themen: |
Camptothecin biosynthesis |
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| Anmerkungen: |
Date Revised 22.03.2024 published: Print-Electronic Citation Status Publisher |
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| doi: |
10.1111/jipb.13649 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM370066316 |
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| 520 | |a © 2024 Institute of Botany, Chinese Academy of Sciences. | ||
| 520 | |a Camptothecin is a complex monoterpenoid indole alkaloid with remarkable antitumor activity. Given that two C-10 modified camptothecin derivatives, topotecan and irinotecan, have been approved as potent anticancer agents, there is a critical need for methods to access other aromatic ring-functionalized congeners (e.g., C-9, C-10, etc.). However, contemporary methods for chemical oxidation are generally harsh and low-yielding when applied to the camptothecin scaffold, thereby limiting the development of modified derivatives. Reported herein, we have identified four tailoring enzymes responsible for C-9 modifications of camptothecin from Nothapodytes tomentosa, via metabolomic and transcriptomic analysis. These consist of a cytochrome P450 (NtCPT9H) which catalyzes the regioselective oxidation of camptothecin to 9-hydroxycamptothecin, as well as two methyltransferases (NtOMT1/2, converting 9-hydroxycamptothecin to 9-methoxycamptothecin), and a uridine diphosphate-glycosyltransferase (NtUGT5, decorating 9-hydroxycamptothecin to 9-β-D-glucosyloxycamptothecin). Importantly, the critical residues that contribute to the specific catalytic activity of NtCPT9H have been elucidated through molecular docking and mutagenesis experiments. This work provides a genetic basis for producing camptothecin derivatives through metabolic engineering. This will hasten the discovery of novel C-9 modified camptothecin derivatives, with profound implications for pharmaceutical manufacture | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Nothapodytes tomentosa | |
| 650 | 4 | |a camptothecin biosynthesis | |
| 650 | 4 | |a enzymatic assay | |
| 650 | 4 | |a tailoring enzyme | |
| 650 | 4 | |a transcriptomic analysis | |
| 700 | 1 | |a Huang, Jian-Ping |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Yong-Jiang |e verfasserin |4 aut | |
| 700 | 1 | |a Tu, Meng-Ling |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Junheng |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Bingyan |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Guoqing |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Jing |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Sheng-Xiong |e verfasserin |4 aut | |
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