Bee venom contains a variety of toxic components, including vitellogenin, which display various biological, toxicological,and pharmacological activities. However, the biological actions of vitellogenin, a venom protein in bee venom, remainlargely unknown. Here, we demonstrate that Asiatic honeybee (Apis cerana) venom vitellogenin (AcV-Vg) exhibits anti-oxidantand anti-microbial activities. AcV-Vg is expressed in the venom gland and is then secreted into venom. The recombinantAcV-Vg protein was produced in baculovirus-infected insect cells. We found that AcV-Vg reduced cytotoxicity and oxidativedamage against oxidative stress. Furthermore, AcV-Vg bound to microbial surfaces and induced structural damage in themicrobial cell walls, which, in turn, exhibited anti-microbial activity against bacteria and fungi. Together, our data demonstratedthat the bee venom protein AcV-Vg has multifunctional roles as an anti-oxidant and anti-microbial agent.