OBJECTIVE Feeding ruminants must notice the degradability of feed, especially protein. Microbial rumen requires ammonia from rumen degradable protein (RDP) beside that ruminant require bypass protein or rumen undegradable protein (RUP) and microbial crude protein. The aim of the study was to discover the best RDP:RUP ratio in beef cattle diets commonly used by Indonesian farmers using an in vitro methodology. METHODS Samples of Pennisetum purpureum, Leucaena leucocephala, Indigofera zollingeriana, cassava, maize, palm kernel cake, rice bran, and tofu waste were formulated into dietary treatments (dry matter [DM] basis). All experiments were carried out using a 3×3×2 factorial, randomized block design with three replications. Treatments consisted of three protein levels (12%, 14%, and 16%), two energy levels (65% and 70%), and three RDP:RUP ratio levels (55:45, 60:40, and 65:35). The experimental diets were incubated in vitro using buffered rumen fluid for 48 h at 39°C. After incubation, the supernatants were analyzed to determine pH, ammonia concentration, total volatile fatty acid (VFA), and microbial protein synthesis. The residues were analyzed to determine DM, organic matter, protein, and RUP digestibility. RESULTS Increased protein, energy, and RDP levels increased digestibility, ammonia concentrations, total VFAs, and microbial protein synthesis (p<0.05), while rations with 16% protein lowered these parameters (p<0.05). CONCLUSIONS Increased dietary protein (from 12% to 14% DM), energy (from 65% to 70% DM), and RDP (from 55% to 65% crude protein [CP]) levels increased nutrient digestibility, ammonia concentration, total VFA levels, and microbial protein synthesis. The diet containing 14% DM dietary protein and 70% DM energy, which contained 55%, 60%, or 65% CP RDP optimally increased nutrient digestibility, ammonia concentration, total VFA levels, and microbial protein synthesis. Thus, feed based on these RDP:RUP ratios can optimize ruminant productivity.