In the present study, phenolic compounds from Gomphrena globosa L. (GGLPCs) including total phenols, flavonoids, flavonols, flavanols, and phenolic acids were measured by spectrophotometric method. The antioxidant activities of GGLPCs were assessed by radical scavengings of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline -6-sulfonic acid)-diammonium salt (ABTS) cation, oxygen radical absorbance capacity (ORAC), phosphomolybdenum antioxidative power (PAP), ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) assays. Antimicrobial and enzyme inhibitory activities of GGLPCs in vitro were investigated using micro-plate dilution method and colorimetry, respectively. The results showed that GGLPCs containing abundant phenolic compounds, had obvious antioxidant and antimicrobial activities (especially against Escherichia coli and Listeria monocytogenes), and significant inhibitory effects on lipase, α-amylase, α-glucosidase, and trypsin in vitro. Thus, the results showed that GGLPC may be used as a natural antioxidant and antibacterial agent, as well as drugs for the treatment of hyperlipidemia, diabetes, and inflammation-related diseases.