The dependence on non-renewable resources, particularly fossil fuels, has awaken a growing interest in research of sustainable alternative energy sources, such as bioethanol. The production of bioethanol from lignocellulosic materials comprises three main stages, starting with a pretreatment, followed by an enzymatic hydrolysis step where fermentable sugars are obtained for the final fermentation process. Enzymatic hydrolysis represents an essential step in the bioethanol production, however there are some limitations in it that hinders the process to be economically feasible. Different strategies have been studied to overcome these limitations, including the enzyme recycling and the utilization of high solids concentrations. Several investigations have been carried out in different bioreactor configurations with the aim to obtain higher yields of glucose in the enzymatic hydrolysis stage; however, the commonest are Stirred Tank Bioreactors (STBR) and Membrane Bioreactors (MBR). In general, the key criteria for a bioreactor design include adequate mass transfer, low shear stress, and efficient mixing that allows the appropriated interaction between the substrate and the enzyme. Therefore, this review will address the main aspects to be considered for a bioreactor design, as well as, the operational conditions, some characteristics and mode of operating strategies of the two main bioreactors used in the enzymatic hydrolysis stage. Moreover, two types of pneumatically agitated bioreactors, namely bubble column and gas-lift bioreactors, are discussed as promising alternatives to develop enzymatic saccharification due to their low energy consumption compared with STBR.