In this paper, state and kinetic parameters estimation based on extended Kalman filter (EKF) is proposed. Experimental data from alcoholic fermentation process with immobilized cells is used. The measurements of glucose and ethanol concentration are used as on-line measurements for observers design and biomass concentration is used for results verification. Biomass, substrate and product concentrations inside immobilized compounds are estimated using the proposed algorithm. Monitoring of the kinetic parameters could be used for process control.