The emergence and spread of multi-drug-resistant strains of Staphylococcus aureus, Escherichia coli and Sreptococcus pneumoniae in associated with causing various illness in human beings. There is an urgent need for searching new drugs to treat bacterial infections. Glycosmis pentaphylla Retz. (Rutaceae) is a shrubby plant found all over India, used for the healing of wounds of livestock in Indian folk medicine. The crude leaf extract of G. pentaphylla was used to perform the screening of phytochemical, antioxidant, antibacterial activities and characterize the chemical constituents using Gas Chromatography-Mass Spectroscopy (GC-MS). The phytochemical screening and antioxidant activity was determined by free radical scavenging assay like DPPH and ABTS. The antibacterial activity was evaluated by agar well plate method. The phytochemical test results show the presence of alkaloids, saponins, tannins, phenolics except glycosides and amino acids. The antioxidant result found the ethyl acetate extract shows an excellent free radical scavenging activity of 63.4mg/ GAE and 49.10mg/g in DPPH and ABTS assays, respectively. The GC-MS spectrum reflects the presence of many biologically importance volatile constituents including, Quaterphenyl, Hexamethy and Heptatriactontadien-2-One. The better antibacterial activity was observed in S. aureus (17mm) followed by other organisms. MIC range was reported as 20mg/ml (the lowest concentration) against clinically isolated multi-drug resistant bacteria’s. The outcome of this study suggests that use of G. pentaphylla plant extract is a best candidate for the treatment of infections caused by multi-drug resistant bacteria’s.