A real-time, on-line extended Kalman filter was used to describe and monitor the growth of Escherichia coli on glycerol. The growth of E. coli showed an inhibition kinetics with mu(max)=0.806/h, K(s)=0.68 g/l and K(i)=87.4 g/l. As a feeding strategy, the conventional DO-stat with a DDC-PID control method, in which the dissolved oxygen concentration is maintained at a desired level by varying the substrate feedrate, was employed. The Kalman filter was based on an unstructured mathematical model and on-line measured data. The mathematical model comprised of mass balances of the biomass and substrate as well as kinetic and stoichiometric data which were measured prior to the process. For biomass concentration up to 50 g dry weight/l, the estimation of the process was rather accurate. At higher biomass concentration, product formation, indicated by an intense brown coloring of the fermentation broth, occured. Since the effect of this product on biomass production was not included in the mathematical model, the estimated data diverged from the experimental data at biomass concentrations greater than 50 g dry weight/l.