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Aims Beads containing heat-inactivated bacterial biomaterial (BBBs) were prepared for removal of cypermethrin (CPM) and the conditions for this removal were evaluated and optimized via single-factor coupled orthogonal experiments based on five factors. The adsorption characteristics of BBBs and the binding mechanism were then explored. Methods and Results Results showed that the adsorption rate of CPM could reach 98% with beads prepared under optimized conditions: equal volumes of Lactobacillus cell debris derived from 1×1011 CFU; 2% hydroxypropyl-β-cyclodextrin and 2.5% activated carbon concentration, were mixed to give mixture TM, and this and SA, was mixed 1:4 with sodium alginate (SA) and beads were prepared using a 26-Gauge needle). The best adsorption conditions were initial CPM concentration of 10 mg l-1, incubation time of 24 h, and rotational speed of 180 rpm. BBBs have a well-formed structure and abundant surface functional groups, such as –COOH, –OH, –NH, –CH, –CO, –C=C. The adsorption process conformed to pseudo-second-order kinetic, and it was also a Freundlich monolayer adsorption, and the calculated maximum adsorption capacity was 9.69 mg g-1 under optimized conditions. Conclusions BBBs showed the highest CPM removal capacity and a good tolerance ability. Significance and Impact of the Study Our results provided a theoretical foundation for developing an adsorbent with heat-inactivated Lactobacillus plantarum (L. plantarum) RS60 for removing CPM in wastewater or drinks.