Oxygen requirements and mass transfer in hairy‐root culture

Abstract

Oxygen mass transfer in clumps of Atropa belladonna hairy roots was investigated as a function of root density and external flow conditions. Convection was the dominant mechanism for mass transfer into root clumps 3.5 to 5.0 cm in diameter; Peclet numbers inside the clumps ranged from 1.4 x 10(3) to 7.1 x 10(4) for external superficial flow velocities between 0.4 and 1.4 cm s(-1). Local dissolved-oxygen levels and rates of oxygen uptake were measured in aflow chamber and in bubble column and stirred bioreactors. When air was used as oxygen source, intraclump dissolved-oxygen tensions ranged from90% to 100% air saturation at high external flow velocity andlow root density, to less than 20% air saturation in dense root clumps. Specific oxygen-uptake rate declined with increasing root density. When external boundary layers around individual roots were eliminated byforcing liquid through the clumps at superficial velocities between 0.2 and1.0 cm s(-1), internal dissolved-oxygen tension was maintained at 95% to 100% air saturation and rate of oxygen uptake at 1.6 x 10(-6) g g(-1) s(-1) dry weight. Liquid culture of single A. belladonna hairy roots was used to investigate the effect of dissolved-oxygen tensionon root growth and morphology. Total root length and number of root tips increased exponentially at oxygen tensions between 70% and 100%air saturation. Specific growth rate increased with oxygen tension up to 100% air saturation; this result demonstrates that hairy roots aeratedwithout oxygen supplementation are likely to be oxygenlimited. No growth occurred at 50% air saturation. Growth of hairy roots proceeded with an average length per tip of about 1 cm; this value was essentially independent of dissolved-oxygen tension between 70% and 100% air saturation. (c) 1994 John Wiley & Sons, Inc.