Identification of membrane engineering targets for increased butanol tolerance in Clostridium saccharoperbutylacetonicum

John A. Linney, Sarah J. Routledge, Simon D. Connell, Tony R. Larson, Andrew R. Pitt, Elizabeth R. Jenkinson, Alan D. Goddard*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


There is a growing interest in the use of microbial cell factories to produce butanol, an industrial solvent and platform chemical. Biobutanol can also be used as a biofuel and represents a cleaner and more sustainable alternative to the use of conventional fossil fuels. Solventogenic Clostridia are the most popular microorganisms used due to the native expression of butanol synthesis pathways. A major drawback to the wide scale implementation and development of these technologies is the toxicity of butanol. Various membrane properties and related functions are perturbed by the interaction of butanol with the cell membrane, causing lower yields and higher purification costs. This is ultimately why the technology remains underemployed. This study aimed to develop a deeper understanding of butanol toxicity at the membrane to determine future targets for membrane engineering. Changes to the lipidome in Clostridium saccharoperbutylacetonicum N1–4 (HMT) throughout butanol fermentation were investigated with thin layer chromatography and mass spectrometry. By the end of fermentation, levels of phosphatidylglycerol lipids had increased significantly, suggesting an important role of these lipid species in tolerance to butanol. Using membrane models and in vitro assays to investigate characteristics such as permeability, fluidity, and swelling, it was found that altering the composition of membrane models can convey tolerance to butanol, and that modulating membrane fluidity appears to be a key factor. Data presented here will ultimately help to inform rational strain engineering efforts to produce more robust strains capable of producing higher butanol titres.

Original languageEnglish
Article number184217
JournalBiochimica et Biophysica Acta - Biomembranes
Issue number8
Early online date30 Aug 2023
Publication statusPublished - Dec 2023

Bibliographical note

Funding Information:
We are grateful for funding from Aston University (JL PhD Scholarship) and BBSRC ( BB/S01943X/1 and BB/R02152X/1 – ADG and ARP).

© 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (


  • Atomic force microscopy
  • Butanol toxicity
  • Clostridia
  • In vitro stability assays
  • Lipid membrane
  • Lipidomics


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