In this research, the effects of lyophilized Urtica dioica L. water extract (LUDWE) on the shelf life of vacuum-packaged beef steaks were evaluated. For this purpose, beef steaks were arranged as control (vacuum packaging, VP), 150 ppm LUDWE + VP, 300 ppm LUDWE + VP and 450 ppm LUDWE + VP, and were stored at 2 ± 0.5C for 21 days. During storage, psychrotrophic bacteria, lactic acid bacteria, Pseudomonas, Enterobacteriaceae counts, pH, color and thiobarbituric acid reactive substances (TBARS) values of beef steaks were determined. By adding LUDWE in the beef steaks, TBARS values decreased (P < 0.01), and minimum values were determined in 450 ppm LUDWE + VP groups (P < 0.05). Depending on the increase in the LUDWE level, psychrotrophic (P < 0.01), Pseudomonas (P < 0.01), Enterobacteriaceae (P < 0.01) and lactic acid bacteria (P < 0.01) counts decreased, and the highest counts of these parameters were determined in the control group. LUDWE also had significant effects on L* (P < 0.01), a* (P < 0.01) and b* (P < 0.01) values of vacuum-packaged beef steaks. PRACTICAL APPLICATIONSBeef steak is more susceptible to microbial spoilage and lipolytic reactions; these can lead to a shorter shelf life with potential risks for human health during storage at low temperatures. In beef steaks, lipid oxidation can be decreased and spoilage bacterial counts can be suppressed using natural antioxidants and antimicrobials such as lyophilized Urtica dioica L. water extract (LUDWE). This work clearly showed that LUDWE could be used as a good natural antioxidant and antimicrobial agent in fresh meat products such as beef steaks.