Liu Dongming, Liu Zhuoyuan, Shan Xiaoyu, Wang Lian, Gao Song, Zhou Jingwen
Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
J Agric Food Chem. 2025 Aug 27;73(34):21445-21452. doi: 10.1021/acs.jafc.5c06272. Epub 2025 Aug 18.
Shikimic acid (SA), a critical intermediate in the synthesis of numerous high-value aromatic compounds, is extensively used in the food and pharmaceutical industries. In this study, efficient SA synthesis in was achieved through dynamic and static regulatory strategies. Initial optimization of key enzymes in the SA pathways AroG, AroB, AroD, and AroE and the growth-regulating gene enabled the accumulation of 7.4 g/L SA. Subsequently, the expression of and was adjusted to ensure adequate NADPH supply. The SA sensor module was then optimized to achieve a response range up to 25 g/L SA, and a negative feedback regulation system was constructed and characterized to balance cellular growth with maximal SA production. An accelerated evolution strategy enhanced the strain's tolerance and identified the regulatory factor StpA that confers tolerance through transcriptome analysis. Finally, the tolerant engineered strain SA35 was fermented in a 5 L bioreactor, and the SA titer was 1.3 times higher than that of the control strain SA33, reaching 97.3 g/L, with a glucose conversion rate of 0.32 g/g. Therefore, these methods provide insights for the efficient synthesis of SA and its derivatives.
莽草酸（SA）是众多高价值芳香族化合物合成中的关键中间体，广泛应用于食品和制药行业。在本研究中，通过动态和静态调控策略实现了SA的高效合成。对SA途径中的关键酶AroG、AroB、AroD和AroE以及生长调节基因进行初步优化，使得SA积累量达到7.4 g/L。随后，调整 和 的表达以确保充足的NADPH供应。接着对SA传感器模块进行优化，使其对SA的响应范围高达25 g/L，并构建和表征了负反馈调节系统，以平衡细胞生长与最大SA产量。加速进化策略提高了菌株的耐受性，并通过转录组分析鉴定出赋予耐受性的调节因子StpA。最后，在5 L生物反应器中对耐受性工程菌株SA35进行发酵，SA产量比对照菌株SA33高1.3倍，达到97.3 g/L，葡萄糖转化率为0.32 g/g。因此，这些方法为SA及其衍生物的高效合成提供了思路。
