Hypericum perforatum L. (St John's wort), although primarily utilized in traditional medicine, is also frequently employed in modern therapy. Within the scope of the study, H. perforatum was collected from Kahramanmaraş region and dried. Extracts derived from above-ground parts were analyzed to determine their levels of total phenolic and flavonoid compounds, as well as their antioxidant, anticancer and antimicrobial properties. Following the GC-MS analysis of the extracts, a total of 18 distinct fatty acids were identified. The main fatty acid components were identified as behenic (37.90 %), linoleic (21.22 %), gamma-linolenic (15.87 %), oleic (9.45 %) and palmitic acid (7.64 %). The plant extracts were found to have a total phenolic content of 60.22 mg GAE g-1 and a flavonoid content of 7.68 µg QE g-1, as determined through analysis. Additionally, FRAP and IC50 values were determined as 26.96 µg AAE g-1 and 0.44 µg mL-1, respectively. The antimicrobial activities of H. perforatum extracts were investigated among a total of 11 microorganisms, including 9 bacteria and 2 yeasts. It has been observed that extracts possess significant antimicrobial activity against all tested microorganisms. H. perforatum extracts were observed to have dose-dependent inhibition of all organisms. It was observed that H. perforatum extracts killed cancer cells at concentrations of 0.1 mg mL-1 and above on H1299, MCF-7 and HUVEC cancer cell lines.