Application of membrane tubing aeration and perfluorocarbon to improve oxygen delivery to hairy root cultures

Growth and atropine production by Atropa belladonna hairy roots were studied in bioreactor cultures using porous polypropylene membrane tubing as a supplementary aeration device and with FC-43 perfluorocarbon emulsion added to the medium. Both these treatments were applied to improve oxygen transfer to the roots. Membrane tubing aeration allowed direct delivery of oxygen within the root clump, thus overcoming mass transfer resistances associated with poor intraclump penetration of liquid convective currents. Combined air sparging and membrane tubing aeration in a gas-driven bioreactor supported biomass levels 32-65% higher than sparging only of air or oxygen-enriched air at the same total gas flow rate. The optimal air flow rate ratio between the sparger and membrane tubing giving the maximum final biomass concentration was 0.2:0.4 L min(-1). Intraclump dissolved oxygen tensions at high biomass densities were generally greater using air delivered by combined sparger-membrane tubing aeration than with sparging only of air or oxygen-enriched air. Specific atropine levels were not significantly affected by membrane tubing aeration. Indicators of anaerobic metabolism, such as lactic acid, ethanol, and ADH activity levels, were not significantly different in sparged and membrane-aerated systems; A. belladonna hairy roots also did not produce aerenchyma in response to oxygen limitations. Addition of perfluorocarbon emulsion to Murashige and Skoog medium in sparged stirred tank bioreactors did not improve growth, even when the emulsion was continuously recycled for re-aeration in an external vessel. Perfluorocarbons are associated with enhancement of gas-liquid oxygen transfer, so their ineffectiveness in this work most likely reflects the dominance of liquid-solid transfer resistances in hairy root cultures. The results of this investigation highlight the importance of developing new approaches for site-directed aeration of hairy root cultures, targeting oxygen delivery into the zones of highest root density.