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Efflux pumps for optimising bioproduction from renewable feedstocks
Secondary Supervisor(s): Professor Jessica Blair
University of Registration: University of Birmingham
BBSRC Research Themes: Renewable Resources and Clean Growth (Industrial Biotechnology)
Project Outline
Petrochemical feedstocks derived from crude oil are the basis of many of the materials and products that we use every day, from plastics to fuels. However, these feedstocks and processes are unsustainable and contribute to climate change. Therefore, there is currently a major shift globally to biological processes using renewable feedstocks. These feedstocks and processes increase sustainability and decrease carbon footprint, both critical for combating climate change and building a circular economy.
Bacteria have a major part to play in this drive for sustainability, and many chemicals and products previously made from crude oil-derived feedstocks using chemical processes are now made by bacteria from green feedstocks [Yang et al., 2021].
Bacteria constantly adapt to their environment and respond to stresses and toxic compounds that they encounter. A common mechanism that protects bacteria against a range of toxic molecules (such as organic solvents and antibiotics) are efflux pumps, molecular machines that pump such molecules out of the cell. It is known that efflux pumps make bacteria more tolerant to a range of solvents and other chemicals used in industrial processes [Tsukagoshi, 2000].
We have recently found that there is a fundamental link between the way in which bacteria generate energy (needed for growth, cellular functions, and product generation) and efflux pumps (required for removing harmful chemicals) [Whittle et al. 2024]. Energy is needed to drive efflux pumps, but we have discovered that the energy level within the cell is responsible for regulating fundamental behaviours, and that efflux pumps impact on these behaviours.
We think that this is very important in understanding how bacteria are able to survive in industrial processes and we can maximise product yields.
The main research questions are:
- How is the energy state of the cell linked to efflux pumps?
- How is this link regulated? What are the sensors and regulators involved in this process?
- How does the energy state of the cell impact on organic solvent tolerance?
- How can we use this link to improve bacterial processes that generate useful compounds?
We will use a combination of approaches, grounded in single-cell analysis of bacteria using flow cytometry and microscopy that we have previously developed [Whittle et al. 2019]. This will allow us to measure how individual bacteria respond to stresses, regulate their energy state, and control efflux pumps. We will also use a variety of molecular biology methods to genetically modify bacteria in order to better understand mechanisms of energy metabolism, efflux, organic solvent tolerance, and their regulation.
This interdisciplinary project will be supervised by Dr Tim Overton (School of Chemical Engineering), an expert on single-cell analysis of bacteria, bacterial physiology, and bioprocessing, and Professor Jessica Blair (Department of Microbes, Infection and Microbiomes), an expert on bacterial efflux pumps and antimicrobial resistance.
References
Tsukagoshi N, Aono R. (2000) Entry into and release of solvents by Escherichia coli in an organic-aqueous two-liquid-phase system and substrate specificity of the AcrAB-TolC solvent-extruding pump. J Bacteriol. 182(17):4803-10. doi: 10.1128/JB.182.17.4803-4810.2000.
Whittle EE, Legood SW, Alav I, Dulyayangkul P, Overton TW, Blair JMA. (2019) Flow Cytometric Analysis of Efflux by Dye Accumulation. Front Microbiol. doi: 10.3389/fmicb.2019.02319.
Whittle EE, Orababa O, Osgerby A, Siasat P, Element SJ, Blair JMA, Overton TW. (2024) Efflux pumps mediate changes to fundamental bacterial physiology via membrane potential. mBio0:e02370-24. doi: 10.1128/mbio.02370-24
Yang D, Prabowo CPS, Eun H, Park SY, Cho IJ, Jiao S, Lee SY. (2021) Escherichia coli as a platform microbial host for systems metabolic engineering. Essays Biochem. 65(2):225-246. doi: 10.1042/EBC20200172.