AbstractXylo-oligosaccharides (XOS) are reported to have beneficial health properties, and they are considered to be functional food ingredients. Corn fiber separated from distillers dried grains with solubles (DDGS) could be a valuable feedstock for XOS production. The objective of this study was to determine the efficacy for autohydrolysis to produce XOS using fiber separated from DDGS and to determine the optimum temperature for XOS production. Corn fiber was treated with deionized water in a Parr-reactor, at temperatures ranging from 140 to 220 °C to produce XOS. The maximum total yield of XOS in the solution was 18.6 wt% of the corn fiber at 180 °C.