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Enhancement of solvent tolerance and economical bioethanol production by modulated sigma 54 expression of Pseudomonas putida

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Abstract

Sigma 54 was modulated in Pseudomonas putida to enhance solvent tolerance to increase Pseudomonas putida solvent production. Three modified P. putida S12 strains (ST-54A, B, and C) were produced by knocking out the endogenous rpoN encoding gene (rpoN) at position σ54 and fusing it with three different ribosome-binding site (RBS) sequences with various translation-initiation rates. For 1-h shock testing and 24-h tolerance tests, these strains were exposed to ethanol, n-butanol, or isobutanol at various RBS-based starting translation rates. Under every treatment situation, modified strains outlived the wild-type strain. It was discovered that the concentration of sigma 54 proteins was correlated with the level of solvent tolerance by fusing rpoN with increased green fluorescent protein genes. Genes involved in ethanol synthesis were inserted into the modified strains of P. putida S12 and its wild-type counterpart. This study discovered a link between sigma 54 expression and ethanol yields. All three modified strains exhibited significantly higher ethanol production than the wild-type strain, and ST-54A exhibited the highest ethanol production of 0.79%. This is a crucial stage in the development of biocatalytic cells that can be produced economically.

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Data Availability

The authors declare that the data supporting the findings of this study are available within the paper.

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Acknowledgements

We thank members of Technology Commons, the College of Life Science, and National Taiwan University, for their excellent technical assistance in setting up flow cytometry experiments and data analyses.

Funding

This work was supported by the Ministry of Science and Technology (grant number 108–2313-B-002–056-MY3), Executive Yuan, Taiwan. This work was supported by grant MOST 108–2313-B-002 -056 -MY3 from the National Science and Technology Council, Executive Yuan, Taiwan.

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Authors and Affiliations

  1. Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan

    Ching-Yueh Su, Hui-Hsien Kuo, Jung-Hao Wang, Wen-Yi Kao, Ming-Yan Shen, Hsuan-Chen Wu & Kung-Ta Lee

  2. Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan

    Hui-Hsien Kuo

  3. International Industry and Agriculture Innovation Research Center (IIAR), International College, Maejo University, Chiang Mai, 50290, Thailand

    Prakash Bhuyar

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  1. Ching-Yueh Su
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Contributions

CYS and HHK designed the study, analyzed the data, and wrote the manuscript; JHW, WYK, PB, MYS, and HCW assisted with data analyses and manuscript writing; KTL supervised the study and wrote the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Kung-Ta Lee.

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Su, CY., Kuo, HH., Wang, JH. et al. Enhancement of solvent tolerance and economical bioethanol production by modulated sigma 54 expression of Pseudomonas putida. Biomass Conv. Bioref. (2024). https://doi.org/10.1007/s13399-024-05275-0

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