Glycosylated cannabinoids in Cannabis sativa and enzyme design to modulate their synthesis

• 成果类型: Article
• 署名作者: Pinkas, Zoe; Khersonsky, Olga; Berman, Paula; Kuzmich, Nikolay; Rogachev, Ilana; Fleishman, Sarel J.; Aharoni, Asaph
• 署名单位: Weizmann Institute of Science; Weizmann Institute of Science; Weizmann Institute of Science
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-10746
• DOI: 10.1073/pnas.2515688122
• 发表日期: 2025-09-30
• 关键词: acid; precursor; hemp

摘要

Despite extensive study of its chemical composition and long history of medicinal use, the occurrence of glycosylated cannabinoid derivatives in Cannabis sativa has not been documented to date. Here, we identified glycosylated cannabinoids and their common intermediate olivetolic acid (OA) in various C. sativa tissues and cultivars. We moreover ating activity. Enhancing the water solubility of cannabinoids through glycosylation holds potential for pharmaceutical development and cosmetic applications. However, glycosylation of pathway intermediates such as OA may divert metabolic flux away from cannabinoid production, complicating efforts to engineer glycosylated forms. To resolve this, we applied FuncLib design to an AlphaFold- predicted structure of one of screening, we identified mutants that display increased specificity toward cannabinoid that dictates OA positioning within the active site, thereby altering isomer formation. These findings expand the known repertoire of natural cannabinoids and provide a rare example of crystallography- free enzyme design to improve stability, substrate selectivity, and isomer specificity. Furthermore, this work lays the foundation for the tailored biosynthesis of soluble glycosylated cannabinoids in heterologous systems.