Operational optimization of a turbine blade reactor using macroporous support and its application to rice callus regeneration

Optimal operation condition was investigated for immobilized rice callus culture using a turbine blade reactor (TBR²) with polyurethane foam supports. By using polyurethane foam block as immobilization support, the inhibition of cell growth at a high agitation speed was avoided because the hydrodynamic stress against immobilized cell was probably reduced. Experimental results in each operational condition were assessed by means of rice callus growth, immobilization ratio in TBR and those regeneration frequencies in regeneration culture using solid medium. Concerning with pore size of polyurethane foam and support size, three-millimeter cube support of polyurethane foam with an average pore size of 1.3 mm was the most suitable support. The maximum immobilization ratio was 50% under 5% support volume by volume of growth medium. For improving the immobilization ratio of rice callus in the TBR, the optimum TBR operation and modification were investigated further. By repeating a periodic operation 3 times (agitating at 300 rpm for 5 min and then 50 rpm for 2 min, and then 200 rpm of constant agitation speed during the remaining time), almost all supports could entrap rice callus and homogeneous immobilization was attained. The immobilization ratio was improved as compared with that using a constant operation at 200 rpm. Next, the TBR was modified by setting an air sparger inside the stainless mesh cylinder. In the modified TBR, the floating support by air bubbles was reduced, and the immobilization ratio increased further and reached 86.3% when we increased the support volume to 15% under periodic operation on a daily basis. The regeneration frequency of immobilized callus was also slightly increased by periodic operation and modification of the TBR.