Zhang Yapeng, Liu Chenwen, Li Wenqiang, Ma Zhidong, Lv Bo, Qin Lei, Li Chun
Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Ministry of Industry and Information Technology/Institute of Biochemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing, China.
Department of Chemical Engineering, Tsinghua University, Beijing, China; Key Lab for Industrial Biocatalysis, Ministry of Education, Tsinghua University, Beijing, China.
Metab Eng. 2025 Sep;91:347-355. doi: 10.1016/j.ymben.2025.06.004. Epub 2025 Jun 4.
β-caryophyllene, a plant-derived sesquiterpene, serves as a food flavoring, anti-inflammatory agent, antioxidant, and high-energy fuel source. Extraction of β-caryophyllene from plants is a costly and inefficient process. Therefore, microbial cell factories have been employed for the production of β-caryophyllene. However, the limited yield is insufficient for its industrial application. In this study, we balanced the utilization of cellular resources for growth and production by systematically regulating the sterol synthesis pathway to maximize the synthesis of β-caryophyllene. In the competitive pathways concerning sterol and fatty acid synthesis, genes expression was suppressed by substituting the original promoters with a glucose-sensing promoter P and a sterol synthesis promoter P, respectively. This approach effectively increased the production of β-caryophyllene by 6.8 times, reaching 854.7 mg/L. Engineering glucose-sensing pathway altered the strength of P, resulting in an increase in β-caryophyllene production to 1.25 g/L. The cell growth and β-caryophyllene production were further boosted through diploid fusion, resulting in 21.4 g/L β-caryophyllene in fed-batch fermentation. This represents the highest reported production of β-caryophyllene to date. This study provides a valuable reference for the production of sesquiterpenes in microbial cell factories.
β-石竹烯是一种植物来源的倍半萜烯，可用作食品调味剂、抗炎剂、抗氧化剂和高能量燃料来源。从植物中提取β-石竹烯是一个成本高昂且效率低下的过程。因此，已采用微生物细胞工厂来生产β-石竹烯。然而，有限的产量不足以满足其工业应用需求。在本研究中，我们通过系统调节甾醇合成途径来平衡细胞资源用于生长和生产的利用，以最大限度地合成β-石竹烯。在涉及甾醇和脂肪酸合成的竞争途径中，分别用葡萄糖感应启动子P和甾醇合成启动子P替换原始启动子，从而抑制基因表达。这种方法有效地将β-石竹烯的产量提高了6.8倍，达到854.7毫克/升。构建葡萄糖感应途径改变了启动子P的强度，使β-石竹烯产量提高到1.25克/升。通过二倍体融合进一步促进了细胞生长和β-石竹烯的生产，在补料分批发酵中产生了21.4克/升的β-石竹烯。这是迄今为止报道的β-石竹烯的最高产量。本研究为微生物细胞工厂中倍半萜烯的生产提供了有价值的参考。
