Shenzhen Key Laboratory for the Intelligent Microbial Manufacturing of Medicines, CAS Key Laboratory of Quantitative Engineering Biology, Center for Synthetic Biochemistry, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
Synceres Biosciences (Shenzhen) CO., LTD, China.
Metab Eng. 2023 Nov;80:232-240. doi: 10.1016/j.ymben.2023.10.004. Epub 2023 Oct 26.
Cannabinoids are important therapeutical molecules for human ailments, cancer treatment, and SARS-CoV-2. The central cannabinoid, cannabigerolic acid (CBGA), is generated from geranyl pyrophosphate and olivetolic acid by Cannabis sativa prenyltransferase (CsPT4). Despite efforts to engineer microorganisms such as Saccharomyces cerevisiae (S. cerevisiae) for CBGA production, their titers remain suboptimal because of the low conversion of hexanoate into olivetolic acid and the limited activity and stability of the CsPT4. To address the low hexanoate conversion, we eliminated hexanoate consumption by the beta-oxidation pathway and reduced its incorporation into fatty acids. To address CsPT4 limitations, we expanded the endoplasmic reticulum and fused an auxiliary protein to CsPT4. Consequently, the engineered S. cerevisiae chassis showed a marked improvement of 78.64-fold in CBGA production, reaching a titer of 510.32 ± 10.70 mg l from glucose and hexanoate.
大麻素是治疗人类疾病、癌症治疗和 SARS-CoV-2 的重要治疗分子。中央大麻素大麻萜酚酸（CBGA）由大麻素植物酰基转移酶（CsPT4）从香叶基焦磷酸和橄榄萜酸生成。尽管人们努力设计微生物，如酿酒酵母（S. cerevisiae）来生产 CBGA，但由于己酸向橄榄萜酸的转化率低，以及 CsPT4 的活性和稳定性有限，其产量仍然不理想。为了解决低己酸转化率的问题，我们消除了β-氧化途径对己酸的消耗，并减少了其在脂肪酸中的掺入。为了解决 CsPT4 的限制，我们扩展了内质网并将辅助蛋白融合到 CsPT4 上。因此，经过工程设计的酿酒酵母底盘在 CBGA 生产方面有了显著的提高，从葡萄糖和己酸出发，达到了 510.32±10.70 mg/L 的产量。
