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用于己二酸生产的[代谢工程]

[Metabolic engineering of for adipic acid production].

作者信息

Liu Jie, Gao Cong, Chen Xiulai, Guo Liang, Song Wei, Wu Jing, Wei Wanqing, Liu Jia, Liu Liming

机构信息

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.

Joint Laboratory for International Cooperation in Food Safety, Jiangnan University, Wuxi 214122, Jiangsu, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2023 Jun 25;39(6):2375-2389. doi: 10.13345/j.cjb.220992.

Abstract

Adipic acid is a high-value-added dicarboxylic acid which is primarily used in the production of nylon-66 for manufacturing polyurethane foam and polyester resins. At present, the biosynthesis of adipic acid is hampered by its low production efficiency. By introducing the key enzymes of adipic acid reverse degradation pathway into a succinic acid overproducing strain FMME N-2, an engineered . JL00 capable of producing 0.34 g/L adipic acid was constructed. Subsequently, the expression level of the rate-limiting enzyme was optimized and the adipic acid titer in shake-flask fermentation increased to 0.87 g/L. Moreover, the supply of precursors was balanced by a combinatorial strategy consisting of deletion of , over-expression of , and mutation of , and the adipic acid titer of the resulting . JL12 increased to 1.51 g/L. Finally, the fermentation process was optimized in a 5 L fermenter. After 72 h fed-batch fermentation, adipic acid titer reached 22.3 g/L with a yield of 0.25 g/g and a productivity of 0.31 g/(L·h). This work may serve as a technical reference for the biosynthesis of various dicarboxylic acids.

摘要

己二酸是一种高附加值的二元羧酸,主要用于生产尼龙 - 66,以制造聚氨酯泡沫和聚酯树脂。目前,己二酸的生物合成受到其低生产效率的阻碍。通过将己二酸逆向降解途径的关键酶引入琥珀酸高产菌株FMME N - 2中,构建了一种能够产生0.34 g/L己二酸的工程菌JL00。随后,对限速酶的表达水平进行了优化,摇瓶发酵中己二酸的效价提高到了0.87 g/L。此外,通过由删除、过表达和突变组成的组合策略平衡了前体的供应,所得JL12的己二酸效价提高到了1.51 g/L。最后,在5 L发酵罐中对发酵过程进行了优化。经过72 h的补料分批发酵,己二酸效价达到22.3 g/L,产率为0.25 g/g,生产力为0.31 g/(L·h)。这项工作可为各种二元羧酸的生物合成提供技术参考。

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