The Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
The Key Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
Biotechnol J. 2024 Jan;19(1):e2300324. doi: 10.1002/biot.202300324. Epub 2023 Oct 13.
Cembratriene-ol is a good biodegradable biopesticide ingredient with future potential applications in the field of sustainable agriculture. Cembratriene-ol is a monocyclic diterpenoid compound that is synthesized only in the trichome gland of Nicotiana plants. In this study, geranylgeranyl diphosphate synthase gene ggpps from Taxus canadensis and cbtsΔp were heterologously expressed in Saccharomyces cerevisiae W303-1A to successfully synthesize cembratriene-ol. The titer of cembratriene-ol was increased by 1.84-fold compared to the control by overexpressing the S. cerevisiae bifunctional (2E,6E)-farnesyl diphosphate synthase genes ERG20 and cbtsΔp under one promoter P . The titer of cembratriene-ol in the engineered S. cerevisiae BY4741 was increased by 1.39-fold compared to the engineered S. cerevisiae W303-1A. The titer of cembratriene-ol in the engineered S. cerevisiae BY4741 was increased by 2.22-fold compared to the control by overexpressing ERG20 and cbtsΔp, respectively, using two promoters P . Cembratriene-ol was found to be successfully synthesized via the integrated expression of cbtsΔp, ggpps and ERG20 on the genome of S. cerevisiae BY4741. The titer of cembratriene-ol in S. cerevisiae S25 was further increased by 1.80-fold compared to the control via dynamic control of the squalene synthase gene ERG9. Overexpression of the genes cbtsΔp and ggpps using pY26-GPD-TEF in S. cerevisiae S25 with their integration expression increased the titer of cembratriene-ol by 26.1-fold compared to S. cerevisiae S25. The titer of cembratriene-ol was significantly enhanced by mitochondrial compartmentalized expression of cbtsΔp and ggpps, which was 76.3-fold higher than that of the initial strain constructed. It was indicated that the systematic optimization has great potential in facilitating high-level production of cembratriene-ol production in S. cerevisiae.
金松萜二醇是一种良好的可生物降解的生物农药成分，具有在可持续农业领域的潜在应用前景。金松萜二醇是一种单环二萜化合物，仅在烟草植物的毛状体腺中合成。在这项研究中，从加拿大紫杉中异源表达了金松萜二醇合成酶基因 ggpps 和 cbtsΔp，成功地在酿酒酵母 W303-1A 中合成了金松萜二醇。与对照相比，通过在一个启动子 P 下过表达酿酒酵母双功能（2E，6E）-法尼基二磷酸合酶基因 ERG20 和 cbtsΔp，金松萜二醇的产量增加了 1.84 倍。与工程酿酒酵母 W303-1A 相比，在工程酿酒酵母 BY4741 中，金松萜二醇的产量增加了 1.39 倍。与对照相比，通过在两个启动子 P 下分别过表达 ERG20 和 cbtsΔp，工程酿酒酵母 BY4741 中，金松萜二醇的产量增加了 2.22 倍。通过在酿酒酵母 BY4741 基因组上整合表达 cbtsΔp、ggpps 和 ERG20，成功地合成了金松萜二醇。与对照相比，通过动态控制角鲨烯合酶基因 ERG9，酿酒酵母 S25 中金松萜二醇的产量进一步提高了 1.80 倍。在酿酒酵母 S25 中，使用 pY26-GPD-TEF 过表达 cbtsΔp 和 ggpps 及其整合表达，金松萜二醇的产量比酿酒酵母 S25 提高了 26.1 倍。通过 cbtsΔp 和 ggpps 的线粒体区室化表达显著提高了金松萜二醇的产量，比初始构建体高 76.3 倍。这表明系统优化在促进酿酒酵母中金松萜二醇的高水平生产方面具有巨大潜力。
