Biosensor-based high-throughput screening enabled efficient adipic acid production
Biosensor-based high-throughput screening enabled efficient adipic acid production
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DOI：
10.1007/s00253-023-12669-z
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发表时间：
2023-07
影响因子：
5
通讯作者：
Rui Zhi;Nan Cheng;Guohui Li;Yu Deng
中科院分区：
文献类型：
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作者：
Rui Zhi;Nan Cheng;Guohui Li;Yu Deng
己二酸是一种重要的化工原料，但目前的合成方法对环境污染严重。近年来，随着代谢工程和合成生物学的发展，生物合成己二酸的研究取得了很大进展。然而，遗传异质性导致的产品效价下降在很大程度上限制了己二酸等化学品的工业化。因此，为了克服这一挑战，我们组成型表达了反向己二酸降解途径，设计并优化了己二酸生物传感器，并建立了一个高通量筛选平台，以筛选基于优化的生物传感器的高性能菌株。利用该平台，我们成功筛选出了一株己二酸效价为188.08 mg·L−1的菌株。将筛选平台与发酵条件优化相结合，摇瓶发酵己二酸效价达到531.88 mg·L− 1，比出发菌株提高了18.78倍。最终利用筛选出的高效菌株在5L发酵罐中进行放大发酵，其中己二酸效价达到3.62 g·L−1。总之，本研究中开发的策略被证明是一种潜在的有效方法，可以减少遗传异质性，并有望为帮助建立更有效的工业筛选过程提供指导。关键点·开发了微调己二酸生物传感器。·建立高通量筛选平台，筛选高性能菌株。·在5L发酵罐中，己二酸的效价达到3.62g·L-1。
Adipic acid is an industrially important chemical, but the current approach to synthesize it can be of serious pollution to the environment. Rencently, bio-based production of adipic acid has significantly advanced with the development of metabolic engineering and synthetic biology. However, genetic heterogeneity-caused decrease of product titer has largely limited the industrialization of chemicals like adipic acid. Therefore, in the attempt to overcome this challenge, we constitutively expressed the reverse adipate degradation pathway, designed and optimized an adipic acid biosensor, and established a high-throughput screening platform to screen for high-performance strains based on the optimized biosensor. Using this platform, we successfully screened a strain with an adipic acid titer of 188.08 mg·L−1. Coupling the screening platform with fermentation optimization, the titer of adipic acid reached 531.88 mg·L−1under shake flask fermentation, which achieved an 18.78-fold improvement comparing to the initial strain. Scale-up fermentation in a 5-L fermenter utilizing the screened high-performance strain was eventually conducted, in which the adipic acid titer reached 3.62 g·L−1. Overall, strategies developed in this study proved to be a potentially efficient method in reducing the genetic heterogeneity and was expected to provide guidance in helping to build a more efficient industrial screening process.Key points• Developed a fine-tuned adipic acid biosensor.• Established a high-throughput screening platform to screen high-performance strains.• The titer of adipic acid reached 3.62 g·L−1in a 5-L fermenter.