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Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance

  • Biotechnology Methods - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The economic viability of bio-production processes is often limited by damage to the microbial cell membrane and thus there is a demand for strategies to increase the robustness of the cell membrane. Damage to the microbial membrane is also a common mode of action by antibiotics. Membrane-impermeable DNA-binding dyes are often used to assess membrane integrity in conjunction with flow cytometry. We demonstrate that in situ assessment of the membrane permeability of E. coli to SYTOX Green is consistent with flow cytometry, with the benefit of lower experimental intensity, lower cost, and no need for a priori selection of sampling times. This method is demonstrated by the characterization of four membrane engineering strategies (deletion of aas, deletion of cfa, increased expression of cfa, and deletion of bhsA) for their effect on octanoic acid tolerance, with the finding that deletion of bhsA increased tolerance and substantially decreased membrane leakage.

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Acknowledgements

This work was supported by the United States Department of Agriculture National Institute of Food and Agriculture, Award Number 2017-67021-26137. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank the ISU Flow Cytometry Facility.

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  1. Department of Chemical and Biological Engineering, Iowa State University, Ames, IA, 50011, USA

    Miguel C. Santoscoy & Laura R. Jarboe

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Santoscoy, M.C., Jarboe, L.R. Streamlined assessment of membrane permeability and its application to membrane engineering of Escherichia coli for octanoic acid tolerance. J Ind Microbiol Biotechnol 46, 843–853 (2019). https://doi.org/10.1007/s10295-019-02158-6

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