In this study, the zein extracted from distillers dried grains with solubles (DDGS) is utilized to produce a fat analogue, by employing a simple "reduced-pressure distillation microparticulation" method. The optimal conditions for preparing the fat analogue are as follows: a zein concentration of 3 w/v% (based on 60 v/v% ethanol water), an agitator speed of 200 rpm, a xanthan gum amount of 20 wt% (based on zein mass) and a heating temperature of 50 degrees C. The zein-based fat analogue obtained has a volume weighted mean diameter (D[4, 3]) of about 16 gm, which is then applied in the formulation of mayonnaise, at fat substitution ratios of 0% (full-fat), 20%, 40%, 60%, 80% and 100%, respectively. Based on the appearance, stability, total calorific value, as well as rheological, microstructure and sensory analyses, it may be concluded that the fat substitution ratio of no more than 40% is acceptable. This paper illustrates another high-value added use of zein. (C) 2016 Elsevier Ltd. All rights reserved.