Abstract
(2S)-Naringenin, a dihydro-flavonoid, serves as a crucial precursor for flavonoid synthesis due to its extensive medicinal values and physiological functions. A pathway for the synthesis of (2S)-naringenin from glucose has previously been constructed in Saccharomyces cerevisiae through metabolic engineering. However, this synthetic pathway of (2S)-naringenin is lengthy, and the genes involved in the competitive pathway remain unknown, posing challenges in significantly enhancing (2S)-naringenin production through metabolic modification. To address this issue, a novel high-throughput screening (HTS) method based on color reaction combined with a random mutagenesis method called atmospheric room temperature plasma (ARTP), was established in this study. Through this approach, a mutant (B7-D9) with a higher titer of (2S)-naringenin was obtained from 9600 mutants. Notably, the titer was enhanced by 52.3% and 19.8% in shake flask and 5 L bioreactor respectively. This study demonstrates the successful establishment of an efficient HTS method that can be applied to screen for high-titer producers of (2S)-naringenin, thereby greatly improving screening efficiency and providing new insights and solutions for similar product screenings.
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Data Availability
All data presented in this study is available from the correspoding author upon reasonable request.
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Acknowledgements
This work was supported by the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (32021005), and the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study (Grant No. SN-ZJU-SIAS-0013).
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Q. G. and S. G. designed the study and wrote the manuscript. Q. G., W. Z., and J. Z. critically revised the manuscript. Q. G. performed the experiments and analyzed the results. J. Z. designed and supervised the project. All authors discussed the results and commented on the manuscript.
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Gao, Q., Gao, S., Zeng, W. et al. Enhancing (2S)-naringenin production in Saccharomyces cerevisiae by high-throughput screening method based on ARTP mutagenesis. 3 Biotech 14, 85 (2024). https://doi.org/10.1007/s13205-023-03892-6
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DOI: https://doi.org/10.1007/s13205-023-03892-6