Despite recent progresses in the use of eukaryotic expression system, production of membrane proteins for structural studies still relies on microbial expression systems. In this review, we provide protocols to achieve high level expression of membrane proteins in Escherichia coli, especially using the T7 RNA polymerase based expression system. From the design of the construct, the choice of the appropriate vector-host combination, the assessment of the bacterial fitness, to the selection of bacterial mutant adapted to the production of the target membrane protein, the chapter covers all necessary methods for a rapid optimization of a specific target membrane protein.
View Article and Find Full Text PDFJ Nanobiotechnology
June 2018
Background: Titanium dioxide (TiO) particles are commonly used as a food additive (E171 in the EU) for its whitening and opacifying properties. However, the risk of gut barrier disruption is an increasing concern because of the presence of a nano-sized fraction. Food-grade E171 may interact with mucus, a gut barrier protagonist still poorly explored in food nanotoxicology.
View Article and Find Full Text PDFTitanium dioxide (TiO) is commonly used as a food additive (E171 in the EU) for its whitening and opacifying properties. However, a risk of intestinal barrier disruption, including dysbiosis of the gut microbiota, is increasingly suspected because of the presence of a nano-sized fraction in this additive. We hypothesized that food-grade E171 and Aeroxyde P25 (identical to the NM-105 OECD reference nanomaterial in the European Union Joint Research Centre) interact with both commensal intestinal bacteria and transient food-borne bacteria under non-UV-irradiated conditions.
View Article and Find Full Text PDFOrganisms that can withstand anhydrobiosis possess the unique ability to temporarily and reversibly suspend their metabolism for the periods when they live in a dehydrated state. However, the mechanisms underlying the cell's ability to tolerate dehydration are far from being fully understood. The objective of this study was to highlight, for the first time, the cellular damage to Yarrowia lipolytica as a result of dehydration induced by drying/rehydration and freezing/thawing.
View Article and Find Full Text PDFIn this work, the ozone inactivation of resistant microorganisms is studied and a method to assess the efficiency of a drinking water plant to inactivate resistant microorganisms using ozone is proposed. This method aims at computing the fraction of resistant microorganisms that are not inactivated at the exit of an ozonation step by evaluating the duration of the lag phase of the ozone inactivation of these microorganisms and the contact time distribution of these microorganisms with the ozone in the step. To evaluate the duration of the lag phase of the ozone inactivation of resistant pathogenic microorganisms, an experimental procedure is proposed and applied to Bacillus subtilis spores.
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