The necessity to generate advances in the comprehension of modeling and simulation of enzymatic hydrolysis of lignocellulosic materials, based on experimentation, has attracted significant attention currently. This paper proposes a methodology for modeling and parameter estimation of the enzymatic hydrolysis process for sugarcane bagasse. The methodology is composed of three steps: experimental process, approach to the phenomenologically based semi-physical model, and parameter estimation. Experimentally, pretreated sugar cane bagasse was characterized, with 22.60, 13.39, and 23.71% for cellulose, hemicellulose, and lignin, respectively. The enzymatic hydrolysis of cellulose, cellobiose, and glucose concentrations was monitored using a commercial cellulase with an enzymatic activity of 96FPU/mL at 50 °C, pH 4.8, 130 rpm for 74 h. A model was proposed, which includes adsorption effects of the enzymatic complex, pH, temperature, transglycosylation, appearance of new cellulose, and product inhibition. Finally, a global mean percentage squared error cost function (% MSEGlobal) was used, adjusting the glucose, cellobiose, and cellulose variables from the estimation of 29 kinetic parameters. It was possible to obtain a % MSEGlobal of 6.33%, which is lower than the proposed acceptable error of 10%. Thus, the methodology for modeling and parameter estimation achieved good results in this first approximation from experimental development and the formulation of a more complex model.