Cyperus rotundus (Family Cyperaceae) is used as a natural drug with low cost by different local tribes in the Middle East. In this study, the antioxidant and antimicrobial activity of the fermented C. rotundus extract was determined using different assays including DPPH and FRAP, and antibacterial activity. The 1H-NMR analysis was carried out for the profiling of bioactive metabolites in the fermented C. rotundus extract. The results demonstrated strong antioxidant activity for DPPH (49.08 µg/mL) and for FRAP (46.95 mmol Fe (II)/g). The 1H-NMR–based metabolomics analysis showed the presence of 19 metabolites including lactic acid, alanine, acetic acid, succinic acid, pyruvic acid, citric acid, choline, beta glucose, betaine, acetoacetate, caffeine, lysine, ascorbic acid, fructose, xylose, alpha Glucose, sucrose, anserine and fumaric acid. The identified metabolites were in correlation to the antioxidant and antimicrobial activity measured. This study indicated the high potential of the C. rotundus fermented extract for natural remedy applications.