Chen Ying, Su Liqiu, Liu Qi, Zhang Ge, Chen Hongyang, Wang Qinhong, Jia Kaizhi, Dai Zongjie
Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Hubei University of Technology, Wuhan, China.
Key Laboratory of Engineering Biology for Low-carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Biotechnol J. 2025 Jan;20(1):e202400669. doi: 10.1002/biot.202400669.
The sesquiterpene (+)-valencene, with its flavor and diverse biological functions, holds promise for applications in the food, fragrance, and pharmaceutical industries. However, the low concentration in nature and high cost of extraction limit its application. This study aimed to construct a microbial cell factory to efficiently produce (+)-valencene. The strain Yarrowia lipolytica YL238, possessing a stronger capacity for (+)-valencene synthesis, was selected and utilized as the chassis for further modifications. By fine-tuning the mevalonate and squalene synthesis pathways we achieved a remarkable 13.2-fold increase in (+)-valencene titer compared to the original strain. Following directed evolution was employed to screen for efficient (+)-valencene synthase, which further enhanced (+)-valencene production by 138%. Consequently, the engineered strain overproduced 813 mg/L of (+)-valencene in shake flasks, marking the highest titer reported in microbials to date. Furthermore, in fed-batch fermentation, this engineered strain showed the capacity to produce 3.3 g/L of (+)-valencene. This study offers a successful model for the application of the "strain-pathway-enzyme" triune strategy in the metabolic engineering of Y. lipolytica, and these methodologies could be broadly utilized for the synthesis of other natural terpenes.
倍半萜烯（+）-瓦伦烯具有独特风味和多种生物学功能，在食品、香料和制药行业具有应用前景。然而，其在自然界中的低浓度以及高昂的提取成本限制了它的应用。本研究旨在构建一个微生物细胞工厂以高效生产（+）-瓦伦烯。筛选出具有较强（+）-瓦伦烯合成能力的解脂耶氏酵母菌株YL238，并将其作为底盘进行进一步改造。通过微调甲羟戊酸和角鲨烯合成途径，与原始菌株相比，（+）-瓦伦烯产量显著提高了13.2倍。随后采用定向进化筛选高效的（+）-瓦伦烯合酶，这进一步使（+）-瓦伦烯产量提高了138%。因此，工程菌株在摇瓶中过量生产了813mg/L的（+）-瓦伦烯，这是迄今为止微生物中报道的最高产量。此外，在补料分批发酵中，该工程菌株能够生产3.3g/L的（+）-瓦伦烯。本研究为“菌株-途径-酶”三位一体策略在解脂耶氏酵母代谢工程中的应用提供了一个成功范例，这些方法可广泛用于合成其他天然萜类化合物。
