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Delivering fibers more distally could be important to prevent or treat colonic diseases such as cancer and ulcerative colitis. Here, we hypothesized that fermentation of fast-fermenting soluble fibers by the colonic microbiota is delayed when they are presented in a mixture due to hierarchical utilization of fibers. A series of in vitro fermentation studies was performed using fecal microbiota obtained from three healthy donors using single dietary fibers [arabinoxylan, chondroitin sulfate (CS), galactomannan (GM), polygalacturonic acid (PGA), xyloglucan (XG)] and a mixture containing an equal amount of each. Substrate disappearance analysis, as measured by GC–MS, revealed that CS, PGA, and XG utilization was delayed when present in the mixture. 16S rRNA sequencing showed certain fibers consistently increased specific genera in the microbiota of all donor groups. Mixing different types of fermentable dietary fibers might be a logical strategy for delivering fibers into more distal regions of the colon.