This study examined the effects of recombinant bacterial expansin and exogenous xylanase and cellulase enzyme preparation (EFE) on preingestive hydrolysis, in vitro ruminal fermentation, and sugar profile of bermudagrass silage. In Experiment 1, the effects of 2 levels of EFE (0, 2.33 mg/g) and 4 levels of expansins (0, 304, 616, 888 μg/g) were evaluated using an in vitro batch culture of buffered-rumen fluid. Dried ground bermudagrass silage (0.50 g; 1 mm) was used as substrate. Treatments were arranged in a 2 × 4 factorial using a randomized block design. Samples were incubated for 24 h at 39°C in quadruplicate per run in 3 independent runs. Gas production was measured at 0, 3, 6, 12 and 24 h. In Experiment 2, preingestive effects of EFE and expansins were evaluated by incubating the substrate in deionized water containing 0.02% (vol/vol) sodium azide. Samples were incubated for 24 h at room temperature (25°C) in quadruplicate in 2 independent runs. Data from both experiments were analyzed using NLME package of R. Model included the effects of EFE, Dose and run was used as a blocking factor, repeated statement was used to estimate gas production. In Experiment 1, EFE and high dose of expansins synergistically increased gas production at 3, 6, 12, and 24 h compared with EFE only (56.2 vs 54.7 mL/ g OM P < 0.01). Similarly, synergistic increases in digestibility of DM (47.5 vs 43.9% P = 0.04), NDF (33.9 vs 30.7% P = 0.08) and ADF (27.3 vs 23.8% P = 0.09) were detected. Adding EFE alone increased total VFA concentration (71.4 vs 63.2 mM P < 0.01) and decreased total CH₄ production while adding expansin alone decreased acetate-to-propionate ratio (2.62 vs 2.67 P < 0.02). In Experiment 2, EFE and expansins synergistically increased % losses of DM, and NDF, regardless of the expansin dose (P < 0.05). Similarly, compared with EFE alone, synergistic increases in release of total reducing sugar (42.5 vs 35.3 mg/ g P = 0.04) and cellobiose (1.74 vs 1.29 mg/mL P = 0.05) were detected. In conclusion, the combination of EFE and expansins synergistically increased NDF hydrolysis and DM and digestibility of bermudagrass silage.