450690 Non-Standard Amino Acid Labeling of the Extracellular Matrix of Probiotics for In Vivo Imaging of the Gut
Here we present the engineering of a probiotic strain, Escherichia coli Nissle 1917 (ECN), to produce curli nanofibers containing NSAA that can participate in click chemistry reactions. Curli nanofibers are part of the extracellular matrix of ECN, and contribute to the bacterial adhesion in the gut and to biofilm formation. In vitro experiments demonstrate that ECN can effectively incorporate NSAA into proteins without recoding of the ECN genome, and only with minimal effects on the fitness of the bacteria. Subsequent dye binding via click chemistry allows for the selective labeling of curli nanofibers containing NSAA. During in vivo experiments, after colonization of the gut and expression of the curli nanofibers in the presence of NSAA, fluorescent labels can be used to detect the presence over time and the spatial distribution of ECN bacteria in the gastrointestinal tract. At a longer term, our technology could be optimized for triggered expression of NSAA-containing curli nanofibers at specific locations in the gut where disease markers are produced.
See more of this Group/Topical: Nanoscale Science and Engineering Forum