Garcinia gummi-gutta L. or better known as Malabar tamarind, belongs to the family of Clusiaceae. The plant is widely used for folk medicine and food preparation. The present study aimed to examine the chemical composition, anti-microbial, and cytotoxic activities of essential oil extracted from the leaf of G. gummi-gutta (EOG). EOG obtained was analyzed using chromatography techniques and its antimicrobial activity and cytotoxic potential were assessed. A total of nineteen constituents were identified with both β- caryophyllene (53.82 ) and (E)-β-farnesene (22.62 ) predominated in the EOG. In the antimicrobial assay, the EOG exhibited a substantial effect on methicillin-resistant Staphylococcus aureus (MRSA) with MIC of 0.156 μg/mL. On the other hand, in vitro 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that the EOG exhibited a notable cytotoxic effect against human breast cancer MDA-MB-231 and MCF- 7/TAM(R) cells. It is worth to point out that the EOG had a high cytotoxic selectivity toward MDA-MB-231 (4.38) and MCF-7/TAM(R) (4.0) cancer cells, which indicates its potential as therapeutic agents. © 2020 Har Krishan Bhalla & Sons.