Enhanced Anthocyanin Production from Grape Callus in an Air-Lift Type Bioreactor Using a Viscous Additive-Supplemented Medium

Abstract

An N-medium containing carboxymethyl cellulose (CMC) was applied to an air-lift type bioreactor culture of grape (Vitis vinifera cv. Bailey alicant A.) callus, and anthocyanin production was investigated. Grape callus grew well at an air flow rate of 80 ml/min and anthocyanin production was significantly increased in the N-medium, reaching 17 mg/l after 7 d of culture. The anthocyanin content of the N-medium was about two times higher than that of the conventional medium without CMC. The effect of air flow rate was also investigated within the range from 40 to 160 ml/min. A twofold increase in anthocyanin content was obtained at all the air flow rates tested in the N-medium. The distribution of grape callus size obtained after 7 d of the bioreactor culture was investigated. The average callus size was 490 mum which was 1.6 times larger than that obtained in the conventional medium. It was found that large calli with a relatively high anthocyanin pigment content were formed in the bioreactor culture using the N-medium. The fluid dynamics in the bioreactor was also investigated at three points (top, middle and bottom) in the bioreactor by laser doppler velocimetry. The average axial velocity of the circulated medium was 0.4 times lower than that of the conventional medium while their average radial velocities were almost the same (zero). The standard deviation of radial velocity fluctuation in the N-medium was also 0.4 times less than that in the conventional medium. These results suggest that turbulent flow occurred in the bioreactor culture using the conventional medium and the degree of turbulent flow decreased significantly when 0.8% CMC was added to the medium to prepare the N-medium. A change of the flow pattern is considered to be the cause of the decrease in hydrodynamic stress, resulting in enhanced pigment production due to the enlargement of the callus.