Abstract
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−1 under 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 −1 in a 5-L fermenter.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Funding
This work was supported the National Key R&D Program of China (2021YFC2100700), the National Natural Science Foundation of China (21877053, 22008088), the Natural Science Foundation of Jiangsu Province (BK20220089), Jiangsu Planned Projects for Postdoctoral Research Funds (2020Z012), China Postdoctoral Science Foundation (2020M681485, 2021T140277), and Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (TSBICIP-KJGG-015).
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RZ, GL, and YD: conceived, designed the study and experiments. RZ, NC conducted the experiments and analyzed the data. RZ, GL, and YD wrote the manuscript. All authors had read and approved the final manuscript.
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Zhi, R., Cheng, N., Li, G. et al. Biosensor-based high-throughput screening enabled efficient adipic acid production. Appl Microbiol Biotechnol 107, 5427–5438 (2023). https://doi.org/10.1007/s00253-023-12669-z
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DOI: https://doi.org/10.1007/s00253-023-12669-z