This study was conducted to investigate the effects of fermented spent mushroom substrates (F-SMS) of Flammulina velutipes on growth performance, carcass traits, and economic characteristics of Hanwoo steers. A yeast strain (Saccharomyces sp. UJ14) and Bacillus strain (Bacillus sp. UJ03) isolated from fresh spent mushroom substrates of Flammulina velutipes were used as probiotics to prepare F-SMS. Twenty-four Hanwoo steers (14 months old) were allocated to three dietary treatments via a randomized block design and were slaughtered at 30 months of age. These treatment groups included Control (TMR), T1 (TMR containing 10% of F-SMS) group, and T2 (TMR containing 30% of F-SMS). Body weight gain was not influenced by the experimental diets. DM and TDN intakes in the finishing period were significantly (p < 0.05) greater in group T1 than in other groups. CP intake was significantly (p < 0.05) greater in group T2 than in other groups during the whole experimental period. Among carcass traits, rib-eye area and back fat thickness tended to increase with F-SMS supplementation. The appearance rate (%) of a meat yield more than grade A was the highest in group T1. The net profits increased by 1.2% and 13.3% in groups T1 and T2, respectively. In conclusion, if a proper feeding program (including feed safety) can be ensured, spent mushroom substrates of Flammulina velutipes can prove to be a highly profitable feed source for Hanwoo steers.