Chen Jiang, Huang Longzheng, Ye Bang-Ce, Zhou Ying
Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China.
Microb Cell Fact. 2025 May 16;24(1):110. doi: 10.1186/s12934-025-02744-7.
Sclareol, a diterpene alcohol derived from Salvia sclarea, is primarily used in the synthesis of ambrox, an alternative to the expensive spice ambergris. However, commercial production of sclareol from plant extraction is costly and environmentally problematic, limiting its scalability. Recent advances in synthetic biology have enabled the construction of efficient cell factories for sclareol synthesis, offering a more sustainable solution.
In this study, we engineered Yarrowia lipolytica to produce sclareol by integrating genes encoding (13E)-8α-hydroxylabden-15-yl diphosphate synthase (LPPS) and sclareol synthase (SCS). Sclareol titers were further enhanced through the fusion of SsSCS and SsLPPS proteins, as well as multi-copy gene integration. To increase the precursor geranylgeranyl diphosphate (GGPP), we overexpressed various geranylgeranyl diphosphate synthases (GGS1), resulting in significant accumulation of GGPP. Additionally, optimization of the mevalonate pathway, coupled with the downregulation of lipid synthesis and upregulation of lipid degradation, directed more acetyl CoA towards sclareol production.
In this study, we reprogrammed the metabolism of Y. lipolytica by combinatorial metabolic engineering with a sclareol titer of 2656.20 ± 91.30 mg/L in shake flasks. Our findings provide a viable strategy for utilizing Y. lipolytica as a microbial cell factory to produce sclareol.
香紫苏醇是一种从南欧丹参中提取的二萜醇，主要用于合成龙涎醚，后者可替代昂贵的香料龙涎香。然而，通过植物提取进行香紫苏醇的商业化生产成本高昂且存在环境问题，限制了其可扩展性。合成生物学的最新进展使得构建用于香紫苏醇合成的高效细胞工厂成为可能，提供了一种更具可持续性的解决方案。
在本研究中，我们通过整合编码（13E）-8α-羟基赖百当-15-基二磷酸合酶（LPPS）和香紫苏醇合酶（SCS）的基因，对解脂耶氏酵母进行工程改造以生产香紫苏醇。通过融合SsSCS和SsLPPS蛋白以及多拷贝基因整合，进一步提高了香紫苏醇的产量。为了增加前体香叶基香叶基二磷酸（GGPP），我们过表达了各种香叶基香叶基二磷酸合酶（GGS1），导致GGPP显著积累。此外，对甲羟戊酸途径的优化，结合脂质合成的下调和脂质降解的上调，使更多的乙酰辅酶A导向香紫苏醇的生产。
在本研究中，我们通过组合代谢工程对解脂耶氏酵母的代谢进行了重新编程，在摇瓶中香紫苏醇产量达到2656.20±91.30mg/L。我们的研究结果为利用解脂耶氏酵母作为微生物细胞工厂生产香紫苏醇提供了可行的策略。
