SUMMARY This study evaluated the effects of eight combinations of three exogenous enzymes with fibrolytic activity (FIB), amylolytic activity (AMZ), and proteolytic activity (PRO) on the accumulated gas production (PG) and kinetic parameters in simulated fermentations in two diet production systems using the in vitro gas production technique. In experiment 1, ruminal fluid was obtained from two Santa Ines sheep, cannulated in the rumen, kept in a feedlot and fed a diet containing a roughage:concentrate ratio of 20:80. In experiment 2, the ruminal fluid was obtained from two Santa Ines sheep, cannulated in the rumen, and kept on pasture (Brachiaria brizantha cv. Marandu). The accumulated gas production was measured during a 96-hour incubation, measured at 18 different time points. After incubation, the pH, dry matter degradability, organic matter in vitro digestibility, digestibility of neutral detergent insoluble fiber, metabolizable energy, partitioning factor, gas yield, short chain fatty acids and microbial protein production were determined. In the in vitro fermentation using the feedlot diet, the increased use of fibrolytic enzymes increased PG and improved the profile characteristics and kinetic parameters of the fermentation, regardless of the combination with other enzymes. In the in vitro fermentation using the forage diet, increased fibrolytic enzymes also increased PG, with improvements in the kinetic parameters and the profile of the fermentation, independent of the use of the other enzymes. The addition only of fibrolytic enzymes has major potential to optimize the use of feedlot diets as well of forage in ruminant feed.