The objective of this study was to determine the application of citric waste fermented yeast waste (CWYW) obtained from an agro-industrial by-product as a protein source to replace soybean meal (SBM) in a concentrate diet. We also determined the effect of various roughage to concentrate ratios (R:C) on the gas production kinetics, ruminal characteristics, and in vitro digestibility using an in vitro gas production technique. The experiment design was a 3 × 5 factorial design arranged in a completely randomized design (CRD), with three replicates. There were three R:C ratios (60:40, 50:50, and 40:60) and five replacing SBM with CWYW (SBM:CWYW) ratios (100:0, 75:25, 50:50, 25:75, and 0:100). The CWYW product’s crude protein (CP) content was 535 g/kg dry matter (DM). There was no interaction effect between R:C ratios and SBM:CWYW ratios for all parameters observed (p > 0.05). The SBM:CWYW ratio did not affect the kinetics and the cumulative amount of gas. However, the gas potential extent and cumulative production of gas were increased with the R:C ratio of 40:60, and the values were about 74.9 and 75.0 mL/0.5 g, respectively (p < 0.01). The replacement of SBM by CWYW at up to 75% did not alter in vitro dry matter digestibility (IVDMD), but 100% CWYW replacement significantly reduced (p < 0.05) IVDMD at 24 h of incubation and the mean value. In addition, IVDMD at 12 h and 24 h of incubation and the mean value were significantly increased with the R:C ratio of 40:60 (p < 0.01). The SBM:CWYW ratio did not change the ruminal pH and population of protozoa (p > 0.05). The ruminal pH was reduced at the R:C ratio of 40:60 (p < 0.01), whereas the protozoal population at 4 h was increased (p < 0.05). The SBM:CWYW ratio did not impact the in vitro volatile fatty acid (VFA) profile (p > 0.05). However, the total VFA, and propionate (C3) concentration were significantly increased (p < 0.01) by the R:C ratio of 40:60. In conclusion, the replacement of SBM by 75% CWYW did not show any negative impact on parameters observed, and the R:C ratio of 40:60 enhanced the gas kinetics, digestibility, VFA, and C3 concentration.