Aim:The study was conducted to evaluate the levels of exogenous fibrolytic enzymes (EFE) on in vitro digestibilities of dry matter (DM) and organic matter (OM), total gas production (TGP), metabolizable energy (ME) content, and microbial biomass production (MBP).Materials and Methods:The total mixed ration (TMR) was prepared using 30% each of sorghum hay and groundnut straw and 40% compound concentrate mixture to meet nutritional requirement of cow (500 kg) producing 12 kg fat corrected milk. The EFE was incorporated at 0, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, and 400 mg/kg TMR. The TMR substrates with different levels of EFE were in vitro incubated to ascertain their effect on digestibility, gas production, and nutritive values.Results:The significantly (p<0.05) higher and optimum in vitro digestibilities of DM (63.03%) and OM (63.62%) as well as TGP (72.35 ml/500 mg TMR) were observed at supplementation of 240 mg EFE/kg TMR, while ME (7.16 MJ/kg DM) and MBP (97.63 mg/500 mg TMR) were also better.Conclusion:The incorporation of EFE at 240 mg/kg TMR resulted significantly (p<0.05) higher and optimum in vitro digestibilities of DM and OM. The TGP, ME, and MBP were also better. The levels of EFE 240 mg/kg TMR were found suitable for further in vivo study in crossbred cows.