Approaches for Cannabinoid Glycosylation Catalyzed by CrUGT74AN3 and BlCGTase

IF 3.2 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Biotechnology Journal Pub Date : 2025-05-13 DOI:10.1002/biot.70007

Christina Schmidt, Astrid Maria Imann, Nikolay Vasilev, Oliver Kayser

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Abstract

Phytocannabinoids are natural products with highly promising pharmaceutical potential, mainly known from the plant Cannabis sativa. However, their bioavailability is limited due to their high lipophilicity. Modification through glycosylation is known to improve the water solubility and stability of molecules. Enzymatic glycosylation requires specific enzymes with high catalytic activity in combination with efficient production systems. To date, only a few glycosyltransferases with activity toward cannabinoids have been described. In this study, we explore the substrate spectrum of the promiscuous UDP-glycosyltransferase CrUGT74AN3 from Catharanthus roseus and demonstrate activity towards a broad range of cannabinoids and their biosynthetic intermediates. The highest activity was observed using cannabidiol (CBD) as an acceptor molecule. In addition, we show efficient biotransformation of CBD in an engineered Saccharomyces cerevisiae strain. We investigate the influence of the hydrolytic activity of endogenous glucosidases and identify the UDP-glucose supply as a limiting factor in the yeast system. The co-expression of CrUGT74AN3 and a cyclodextrin glycosyltransferase from Bacillus licheniformis in the engineered yeast strain led to the production of CBD-glycosides with up to six glucose moieties from CBD and cyclodextrin in vivo. Finally, we confirm the applicability of the engineered yeast systems to other cannabinoids using cannabigerol and cannabinol.

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CrUGT74AN3和BlCGTase催化大麻素糖基化的研究进展

植物大麻素是一种天然产物，具有很高的药用潜力，主要来自植物大麻。然而，由于它们的高亲脂性，它们的生物利用度有限。已知通过糖基化修饰可以改善分子的水溶性和稳定性。酶糖基化需要具有高催化活性的特定酶与高效的生产系统相结合。迄今为止，只有少数对大麻素具有活性的糖基转移酶被描述。在这项研究中，我们探索了来自Catharanthus roseus的混杂udp -糖基转移酶CrUGT74AN3的底物光谱，并证明了其对多种大麻素及其生物合成中间体的活性。以大麻二酚（CBD）为受体分子，其活性最高。此外，我们还展示了CBD在工程酿酒酵母菌株中的高效生物转化。我们研究了内源性葡萄糖苷酶水解活性的影响，并确定了酵母系统中udp -葡萄糖供应是一个限制因素。在工程酵母菌株中，地衣芽孢杆菌的CrUGT74AN3和环糊精糖基转移酶的共同表达导致CBD和环糊精在体内产生含有多达6个葡萄糖片段的CBD糖苷。最后，我们证实了工程酵母系统对使用大麻二酚和大麻酚的其他大麻素的适用性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine

CiteScore

8.90

自引率

2.10%

发文量

123

审稿时长

1.5 months

期刊介绍： Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.