A fermentation system was supplemented with a device for the measurement of the durations of alkali pump feeding for automatic pH control and an A/D convertor for precise monitoring of pH value by computer. A software program was developed to measure buffer capacities from the pH signal and the pH control signal during fermentation. By measuring the buffer capacity on-line, levels of acetic acid were estimated by a software sensor using pH signal in a fermentation process of E.coli growing in a minimal medium. The measured values of acetic acid showed correlation to those of estimated by the software sensor. Lacitic acid production was also successfully estimated by the values of buffer capacities measured on-line.