Forced modulation of biological processes: A review

Cell culture or biochemical processes utilizing free or immobilized cells or enzymes are candidates for periodic operation. Even though these processes are isothermal, inhibition frequently arises giving their kinetics a push–pull character of the Turing type so that excited states or multiplicity are observed. Modulation of an input such as a substrate or a nutrient concentration or the cell environment such as the pH can enhance the rates of biochemical reactions that are occurring. This possibility was realized by explorers of reactor periodic operation in the 1980s. However, the use of rotating disc biological filters for aerobic waste treatment precedes these activities by at least a decade. The biologically active slime on the disc surface is exposed alternately to air and to an aqueous media containing dissolved and/or suspended waste matter. Indeed, periodic variation of reactor volume in a CSTR, which is equivalent to periodic variation of space velocity, was explored in the early 1970s. The application of this was to activated sludge waste treatment. Periodic operation has been used to address the problem of plasmid loss in continuous fermenters to produce genetically altered microorganisms. A twofold increase in the concentration of plasmid bearing cells using square wave modulation of the dilution rate has been found in a simulation using a model validated by experimental data. These results are in agreement with other studies attempting to control species populations in chemostats. Improvement in the rate of fermentation of a substrate to form a metabolite has been addressed by modulation. It has been shown that the rate of xylose utilization by a yeast is affected by a periodic variation of pH. These were lowered, but the metabolite yield, ethanol, remained the same. With secondary metabolites associated with the non-growth period in fermentation, modulation of the growth-limiting substrate concentration fed to immobilized live cells resulted in significantly higher yields. Differences in the effect of cycling on primary and secondary metabolites has been observed in several biochemical systems, such as those used for penicillin and monoclonal antibody production or in citric acid fermentation.