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E.coli and other Gram negative bacteria coat themselves in remarkably stable protein fibres, polymers of around a 100 copies of a single protein, on their cell surface. These fibres are assembled via the well‐recognised chaperone‐usher (CU) pathway.
These highly stable polymers represent an appealing scaffold structure. Replacing a small fragment on this protein, potentially permits display of countless copies of such foreign peptides allowing changes in properties of the cell surface.
One limitation to manipulation of this bacterial coat is that each CU assembly system has evolved to export a specific substrate.
Escherichia coli is a widely used host for production of recombinant proteins. Although high levels of protein expression can be achieved, this is often followed by low-yield down-stream processing needed to extract functional proteins. We have recently developed an approach based on cutting-edge microscopy that allows quantitative observations of single, living E. coli cells during osmotic shocks.
The aromatic acid:H+ symporters (AAHS) are a diverse and widespread transporter family that are responsible for the influx of aromatic acids into bacterial cells. Aromatic acid transport could be exploited in bioremediation, microbial cell factories and in developing novel orthogonal components for synthetic biology. However, we currently lack the fundamental understanding of protein structure and function that will be required to support such applications. Can we develop in vitro methods to study the transport kinetics and substrate specificity of the AAHS proteins?
Metal homeostasis is important in all aspects of life and transport of metals across the membrane is important for the cellular uptake of essential (trace) elements or resistance against toxic levels of metals, which need to be exported. Understanding how metal transporters function or monitoring metal transport is thus vital for many applications. For instance, engineering the metal uptake of microbes has applications in bioremediation, while export of metals is fundamental for the resistance of microbes against metal-based antimicrobials. Metal transport across the lipid membrane can, however, be challenging to monitor experimentally, especially in (semi-) high throughput.
Succinic acid, produced petrochemically, is an important feedstock which acts as a precursor for chemicals found in food, pharmaceuticals, green solvents and biodegradable plastics. The U.S. Department of Energy has reported succinate in the 12 top chemical building blocks that can be manufactured from biomass at a market value of $15 bn pa. Therefore, much effort has been spent recently to develop succinate-producing organisms. Corynebacterium glutamicum bacteria grow rapidly and are already used for industrial production of amino acids. Under oxygen deprivation, it produces organic acids such as lactate, acetate and succinate. Considerable effort has therefore gone to engineer it for succinate production.