In this study, the effect of probiotic supplementation on growth performance, blood metabolites, and meat quality of Hanwoo steer was investigated. A total of 32 Hanwoo steers (15-17 months, average body weight 462±37.9 kg) were randomly allotted to 4 dietary treatments (0, 0.5, 1.0, and 1.5% mixed probiotics), with four Hanwoo steers per pen (two replicates per treatments), and reared for 12 months. There were no differences among treatments in growth performance of Hanwoo steer (P>0.05); however, feed intake decreased linearly with increasing levels of mixed probiotics. Growth hormone and Blood Urea Nitrogen (BUN) levels responded linearly with increasing levels of dietary mixed probiotics (P<0.05), but not insulin and blood glucose did not. In particular, total cholesterol was significantly lower for the 1% mixed probiotic treatment in comparison with that of the other treatments (P<0.05). The pH, Thiobarbituric Acid Reactive Substances (TBARS), cooking loss, and meat color were influenced by increasing levels of mixed probiotics (P<0.05), but the carcass characteristics and shear force were not. Regarding sensory evaluation, the addition of mixed probiotics resulted in significant difference in meat color, tenderness, aroma, off-flavor, juiciness, and marbling score, but not in overall acceptability. In addition, fatty acid profiles indicated no differences between control and mixed probiotic treatments. In conclusion, mixed probiotic treatment at 1% levels can enhance consumer preferences possibly by reducing cholesterol and TBARS.