Distillers' dried grain with solubles (DDGS) is a byproduct of bioethanol fermentation, which uses the dry milling technology for starch-rich grains such as corn, wheat, and barley. The current interest in bioethanol is increasing due to the need for renewable liquid fuels specifically in the transportation sector. Since DDGS is rich in crude protein, fat, fiber, vitamins, and minerals, it is currently used as aquaculture, livestock, and poultry feeds. In recent years, DDGS has been used as feedstock in the production of value-added products via microbial fermentation. Numerous studies reported the production organic acids, methane, biohydrogen, and hydrolytic enzymes using DDGS. While DDGS contains remarkable amounts of macronutrients, pre-treatment of DDGS is required for release of the fermentable sugars. The pre-treatment methods such as chemical, physical, and biological origin are either solely used or combined to obtain maximal yields for different applications. Therefore, this review summarizes some of the most prominent pre-treatment processes generating high fermentable sugar yields for the productions of value-added products in the last 5 years. A special focus has been given to the effect of the variability of DDGS on the final product. Integration of hydrolytic enzyme production with the traditional bioethanol production facilities has been discussed for further improvement of bioethanol, methane, and biohydrogen using DDGS as fermentation feedstock.