Aegerolysin proteins ostreolysin A6 (OlyA6), pleurotolysin A2 (PlyA2) and erylysin A (EryA) produced by the mushroom genus Pleurotus bind strongly to CPE, the main sphingolipid in invertebrate cell membranes. Upon binding, these aegerolysins can recruit a protein partner pleurotolysin B (PlyB) that has a membrane-attack complex/perforin (MACPF) domain to form bicomponent transmembrane pore complexes. OlyA6/PlyB, PlyA2/PlyB and EryA/PlyB complexes show selective toxicity toward two important pests: the western corn rootworm and the Colorado potato beetle. Due to their specific interactions with their membrane lipid receptor CPE, these complexes might represent new candidates for development of biopesticides. In this study, we evaluated the toxicities of these aegerolysin/PlyB complexes to honeybees (Apis mellifera carnica) that are ecologically and economically very important as crop pollinators and honey producers. For this purpose, we separated lipids extracted from honeybee midgut using thin-layer chromatography together with standard lipid CPE. We transferred the separated lipids to PVDF membranes and incubated them with the solution of OlyA6 . These assays confirmed the presence of CPE in honeybee midgut cell membranes as well as binding of OlyA6 to CPE in honeybee midgut lipids. Surface plasmon resonance and calcein release assay showed that the interactions of all three aegerolysins with the nonpolar liposomes made of honeybee midgut lipids were considerably stronger in presence of PlyB, as well as their lytic activity. Compared to OlyA6 and PlyB, EryA showed weaker interaction with these liposomes and consequently a lower lytic activity. Lastly, we investigated the effects of OlyA6/PlyB, PlyA2/PlyB and EryA/PlyB on the feeding and survival rates of honey bees. While EryA/PlyB complex did not affect honeybee survival and feeding rates, OlyA6/PlyB and PlyA2/PlyB were toxic to honeybees with 48 h mean lethal concentrations (LC50) of 0.22 and 0.39 mg/mL, respectively.