The antimutagenic effects of whey, acetone extracts, and protein fractions isolated from milk that had been fermented by Lactobacillus helveticus L89 were investigated using the mutagen 4-nitroquinoline-N'-oxide in the Ames test (Salmonella typhimurium TA 100). Fermented milk significantly inhibited mutagenesis induced by 4-nitroquinoline-N'-oxide. However, milk fermented by a nonproteolytic variant of the same strain showed no inhibitory effects. Results were similar for the whey fractions and acetone extracts of the fermented milks. After fermentation, milk proteins were fractionated by size-exclusion HPLC and were tested for antimutagenicity. The fraction showing the greatest activity was further analyzed by reverse-phase HPLC. Our results indicate that antimutagenic compounds are produced in milk during fermentation by L. helveticus, and the release of peptides is one possible contributing mechanism.