The erosion-protective effect of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is controversial. This study aimed to investigate the ability of CPP-ACP chewing gum to prevent a single event of erosive demineralization in situ. Bovine enamel blocks (n = 120) were randomly assigned to 3 phases according to the baseline surface hardness: phase I (PI) - chewing gum with CPP-ACP, phase II (PII) - chewing gum without CPP-ACP, and control phase (PIII) - salivary effect without stimulation (no gum). Nineteen volunteers participated in this study during 3 crossover phases of 2 h. In PI and PII, the volunteers wore intraoral palatal appliances for 120 min and chewed a unit of the corresponding chewing gum for the final 30 min. In the control phase the volunteers wore the appliance for 2 h, without chewing gum. Immediately after intraoral use, the appliances were extraorally immersed in a cola drink for 5 min to promote erosive demineralization. The percentage of surface hardness loss was calculated. The data were analyzed by ANOVA models and Tukey's test. Lower enamel hardness loss was found after the use of chewing gum with CPP-ACP (PI: 32.7%) and without CPP-ACP (PII: 33.5%) compared to the salivary effect without stimulation (PIII: 39.8%) (p < 0.05). There was no difference between PI and PII (p > 0.05). The results suggest that the use of chewing gum immediately before an erosive demineralization can diminish enamel hardness loss. However, the presence of CPP-ACP in the chewing gum cannot enhance this protective effect.