Abstract
Butanol inhibits bacterial activity by destroying the cell membrane of Clostridium acetobutylicum strains and altering functionality. Butanol toxicity also results in destruction of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS), thereby preventing glucose transport and phosphorylation and inhibiting transmembrane transport and assimilation of sugars, amino acids, and other nutrients. In this study, based on the addition of exogenous butanol, the tangible macro indicators of changes in the carbon ion beam irradiation-mutant Y217 morphology were observed using scanning electron microscopy (SEM). The mutant has lower microbial adhesion to hydrocarbon (MATH) value than C. acetobutylicum ATCC 824 strain. FDA fluorescence intensity and conductivity studies demonstrated the intrinsically low membrane permeability of the mutant membrane, with membrane potential remaining relatively stable. Monounsaturated FAs (MUFAs) accounted for 35.17% of the mutant membrane, and the saturated fatty acids (SFA)/unsaturated fatty acids (UFA) ratio in the mutant cell membrane was 1.65. In addition, we conducted DNA-level analysis of the mutant strain Y217. Expectedly, through screening, we found gene mutant sites encoding membrane-related functions in the mutant, including ATP-binding cassette (ABC) transporter-related genes, predicted membrane proteins, and the PTS transport system. It is noteworthy that an unreported predicted membrane protein (CAC 3309) may be related to changes in mutant cell membrane properties.
Key points
• Mutant Y217 exhibited better membrane integrity and permeability.
• Mutant Y217 was more resistant to butanol toxicity.
• Some membrane-related genes of mutant Y217 were mutated.
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Data availability
The raw sequence data reported in this paper have been deposited in the Genome Sequence Archive (Genomics, Proteomics & Bioinformatics 2017) in the National Genomics Data Center (Nucleic Acids Res 2020), Beijing Institute of Genomics (China National Center for Bioinformation), Chinese Academy of Sciences, under accession number CRA003426 that are publicly accessible at https://bigd.big.ac.cn/gsa.
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Acknowledgements
We sincerely thank the National Laboratory of HIRFL and the National Natural Science Foundation of China for giving us the opportunity to perform this project.
Funding
This study was supported by the National Natural Science Foundation of China (11975284 and 11905265), Science and Technology Service Network Initiative of Chinese Academy of Sciences (KFJ-STS-QYZD-197), Project of Lanzhou Science and Technology 2019-1-39.
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Y.G. and D.L. conceived, designed, and supervised the study. Y.G. and X.Z. wrote the paper. Y.G, C.R.L., and X.P.G. performed the experiments. Y.G. and M.M.Z. analyzed the data. Y.J.L., X.Z., and W.J.L. corrected the manuscript. W.J.L. and D.L. final approval of the version to be published. All authors read and approved the final manuscript.
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Gao, Y., Zhou, X., Zhang, MM. et al. Response characteristics of the membrane integrity and physiological activities of the mutant strain Y217 under exogenous butanol stress. Appl Microbiol Biotechnol 105, 2455–2472 (2021). https://doi.org/10.1007/s00253-021-11174-5
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DOI: https://doi.org/10.1007/s00253-021-11174-5