Abstract
The gut microbiome supplements human metabolism and immunity by catabolizing host-indigestible glycans. The microbiome operates as a community of trillions of bacteria, and we are beginning to understand the contribution of individual species to glycan digestion. This work extends wide-field and super-resolution fluorescence imaging to the complexity of the human gut microbiome. Past in vitro studies have been limited by mono-culture approaches, and previous live-cell fluorescence microscopy has been limited by the oxygen-dependence of traditional fluorescent proteins like GFP. To address these limitations, we have implemented anaerobic live-cell imaging based on the fatty-acid binding protein, UnaG, in the gut bacterium Bacteroides thetaiotaomicron (B. theta). We have also extended our previous work on B. theta starch utilization system (Sus) enzymes to mixed cultures of B. theta and Ruminococcus bromii, a key species in resistant starch degradation, to understand the relationship between the two species in a microbiome. Finally, we are studying the xyloglucan locus in Bacteroidesovatus to compare polysaccharide utilization loci across species.
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