SummaryIn this study, DBP (Di‐n‐butyl phthalate) was used as an example to evaluate the plasticiser‐binding ability of lactic acid bacteria strains (LABs). Three strains of Lactobacillus plantarum 121, Leuconostoc mesenteroides DM12 and Lactobacillus acidophilus NCFM were selected from LABs to test their DBP binding ability in different simulated food matrices. The results indicated that starch, gelatinised starch and glucose could significantly promote the DBP binding efficiencies of LABs. Low‐concentration proteins (<5%) increased the binding rates. However, denatured protein had no significant effect on the DBP binding ability. On the contrary, amino acids reduced the binding rates. AFM detection results indicated that the cell morphology of LAB binding with DBP had no change after treated in gastrointestinal digestive simulation systems. Collectively, the LABs could be employed as a biological detoxification agent for human and animals by eliminating plasticiser from foods, feeds and digestive tract in the future.