Microorganism culture is highly complex due to the different metabolic pathways, which are very complex. A metabolic phenomenon called overflow is a challenge to overcome in automatic control tasks of microorganism cultures. In this study, a nonlinear algorithm by sliding modes (sliding mode nonlinear control, SMNC) is proposed for the robust regulation of a fed-batch bioreactor in the presence of parametric and system perturbations. A control design is obtained using Lyapunov functions by techniques to propose a control law such that it is robust, only the output signals (biomass and volume) are used, and the reaction rates do not have to be wholly known. Therefore, a novel and simple controller capable of solving the above problems is obtained.