Abstract
Acidithiobacillus caldus is a typical extreme acidophile widely used in the biohydrometallurgical industry, which often experiences extreme environmental stress in its natural habitat. Hfq, an RNA-binding protein, typically functions as a global regulator involved in various cellular physiological processes. Yet, the biological functions of Hfq derived from such extreme acidophile have not been extensively investigated. In this study, the recombinant strain Δhfq/Achfq, constructed by CRISPR/Cas9-mediated chromosome integration, fully or partially restored the phenotypic defects caused by hfq deletion in Escherichia coli, including impaired growth performance, abnormal cell morphology, impaired swarming motility, decreased stress resistance, decreased intracellular ATP and free amino acid levels, and attenuated biofilm formation. Particularly noteworthy, the intracellular ATP level and biofilm production of the recombinant strain were increased by 12.2% and 7.0%, respectively, compared to the Δhfq mutant. Transcriptomic analysis revealed that even under heterologous expression, AcHfq exerted global regulatory effects on multiple cellular processes, including metabolism, environmental signal processing, and motility. Finally, we established a potential working model to illustrate the regulatory mechanism of AcHfq in bacterial resistance to environmental stress.
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Acknowledgements
We are grateful to Linzhi Ding for his help in three-dimensional excitation emission matrix (3D-EEM) analysis.
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This study was supported by grants from the National Key Research and Development Program of China (2022YFC3401304/001), the National Natural Science Foundation of China (No. 32371540; 21878128), the funding of Key Laboratory of Industrial Biotechnology, Ministry of Education (KLIBKF202005).
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WH and SF designed the research. WH, XH, HB, and ZC performed the experiments. WH and XH analyzed the data. WH wrote the first draft of the manuscript. SF, HY, and JZ contributed to manuscript revision, read, and approved the submitted version. All authors read and approved the manuscript.
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Hu, W., Huo, X., Bai, H. et al. Insights into the complementation potential of the extreme acidophile’s orthologue in replacing Escherichia coli hfq gene—particularly in bacterial resistance to environmental stress. World J Microbiol Biotechnol 40, 105 (2024). https://doi.org/10.1007/s11274-024-03924-0
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DOI: https://doi.org/10.1007/s11274-024-03924-0