SusE facilitates starch uptake independent of starch binding in B. thetaiotaomicron

Summary The Bacteroides thetaiotaomicron starch utilization system (Sus) is a model system for nutrient acquisition by gut Bacteroidetes, a dominant phylum of gut bacteria. The Sus includes SusCDEFG, which assemble on the cell surface to capture, degrade and import starch. While SusD is an essential starch‐binding protein, the precise role(s) of the partially homologous starch‐binding proteins SusE and SusF has remained elusive. We previously reported that a non‐binding version of SusD (SusD*) supports growth on starch when other members of the multi‐protein complex are present. Here we demonstrate that SusE supports SusD* growth on maltooligosaccharides, and determine the domains of SusE essential for this function. Furthermore, we demonstrate that SusE does not need to bind starch to support growth in the presence of SusD*, suggesting that the assembly of SusCDE is most important for maltooligosaccharide uptake in this context. However, starch binding by proteins SusDEF directs the uptake of maltooligosaccharides of specific lengths, suggesting that these proteins equip the cell to scavenge a range of starch fragments. These data demonstrate that the assembly of core Sus proteins SusCDE is secondary to their glycan binding roles, but glycan binding by Sus proteins may fine tune the selection of glycans from the environment. Human gut Bacteroidetes use multi‐protein cell surface complexes to catabolize numerous dietary polysaccharides that the host cannot digest. Here, we reveal that starch‐binding proteins of the starch utilization system in Bacteroides thetaiotaomicron facilitate glycan import independent of their binding sites and influence the length of glycan preferentially imported by the bacterium.