Measurement of biological parameters during fermentation processes

Efficient fermentation control requires the measurement of biological parameters. Three techniques were tested for monitoring. In the first, the NADH-fluorescence of micro-organisms was measured in batch and in continuous cultures under aerobic and anaerobic conditions, providing information on the metabolic status of the cells. The effects of cell concentration and of different substrates (glucose, ethanol and oxygen) were studied. The second technique is the calorimetric determination of various substrates, such as penicillin or enzymes, by an enzyme/thermistor device. With immobilized penicillin acylase (E.C. 3.5.1.11) or penicillinase (E.C. 3.5.2.6), penicillin was determined selectively in a fermentation broth. The thermistor was also used to measure penicillin acylase activity. The third technique is laser flow cytometry. A commercial double-beam flow cytometry system was used to determine cell size, light scattering and the protein, DNA and RNA contents of single cells. Flow cytometry allows rapid and sensitive control of fermentation processes with genetically modified E. coli 5K (pHM12) cells. The results of monitoring the cell size, light scattering, and protein and DNA contents of different micro-organisms during fermentation are outlined.