Bauhinia ungulata L. species, belongs to the genus Bauhinia, popularly known as pata de vaca, is among the species of medicinal employ in Brazil, used to treat diabetes. The aim of this study was to characterize the chemical composition, antimicrobial activity, toxicity on Artemia salina and acetylcholinesterase enzyme inhibition by essential oil from B. ungulata L. The chromatographic analysis revealed 18 components, the majority were the β-caryophyllene (15.9%), caryophyllene oxide (9.2%), α-humulene (8.1%), epi-γ-eudesmol (7.5%), α-bisabolol (4.7%), copaene (3.5%), nerolidol (3.3%), α-bisabolol oxide B (2.5%), spathulenol (2.1%). The essential oil showed high toxicity compared to the tests with Artemia salina and inhibited 95.96%±0.62 of the acetylcholinesterase enzyme. The microorganisms show antimicrobial inhibition with Candida albicans (85%), Bacillus cereus (65.5%) and Staphylococcus aureus (66.4%), Salmonella typhimurium (68.7%) and Citrobacter freundii (46.1%). The oil showed great potential when tested in bioassays.