• KSBB Journal
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• v.13 no.2
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• pp.203-208
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• 1998

We investigated the effects of dissolved oxygen (D.O.) and fructose (C-source) on cell growth and biosynthesis of cyclosporin A (CyA) produced as a secondary metabolite by a wild-type filamentous fungus, Tolypocladium inflatum. This was performed by controlling the level of D.O. and the residual C-source, as required, through adjustment of medium flow rate, medium concentration and agitation rate in fed-batch cultures. CyA production was furned out to be maximal, when D.O. level was controlled around 10% saturated D.O. and concentration of the C-source was maintained sufficiently low (below 2 g/L) not to cause carbon catabolite repression. Under this culture condition, we obtained the highest values of CyA concentration (507.14 mg/L), Qp (2.11 mg CyA/L/hr), $Y_x/s$ (0.49 g DCW/g fructose), $Y_p/s$<(22.56 mg CyA/g fructose), and YTEX>$_p/x$ (48.31 mg CyA/g DCW), but relatively lower values of cell concentration (11.98 g DCW/L) and cell productivity (0.043 g DCW/L/hr), in comparison with other parallel fed-batch fermentation conditions. These results implied that, in the carbon-limited culture with 10% saturated D.O. level, the producer microorganism utilized the C-source more efficiently for secondary metabolism.

• KSBB Journal
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• v.9 no.2
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• pp.211-223
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• 1994

In fermentations with a 4-liter stirred tank bioreactor, a better than two-fold enhancement of the gas-liquid mass transfer coefficient$(k_La)$ in the celite-immobilized fungal cultures of Tolypocladium in flatum over the parallel conventional free-cell was observed at identical biomass concentrations, despite the higher specific oxygen uptake rate of the immobilized fungi during exponential growth. As a result oxygen sufficient conditions, i. e., dissolve oxygen(D.O.) concentrations exceeding 75% air saturation, could be maintained throughout exponential growth period of the immobilized culture, in contrast to the suspended fungal culture, whose D.O. levels fell below 50% air saturation. A linear monotonic dependence of $k_La$ upon impeller agitaion rate was found for both immobilized and conventional cultivation modes over a range of 250 to 550rpm, the slope being a function of biomass concentration for the free but not for the immobilized cell system In contrasts oxygen transfer rate was a much weaker function of aeration rate up to about 2.5 vvm for both culture configurations. Above this level, aeration rate had no further effect on the mass transfer. In addition, the immobilized cultures sustained good morphological and physiological states, leading to almost two times higher cyclosporln A (CyA) productivity overt the parallel free cell system. These experiments suggest that the celite-immobilized fungal system in a stirred tank reactor has considerable promise for scaling up cyclosporin A production in terms of high-density cultivation.

• KSBB Journal
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• v.11 no.1
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• pp.22-29
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• 1996

Immobilized bioprocess was carried out for continuous production of cyclosporin A (CyA) produced intracellularly as a secondary metabolite by a filamentous fungus, Tolypocladium inflatum. Immobilization procedure for entrapping conidiospores of the producer was significantly simplified by use of a modified immobilization technique. A newly-designed immobilized perfusion reactor system (IPRS) showed good process benefits as demonstrated by the role of the high density immobilized cells as an efficient biomass generator, continuously supplying highly active CyA-producing free cells (1.0g/$\ell$/hr) even at very high dilution rate ($0.1hr^{-1}$). IPRS bioprocess was possible since efficient decantor system developed in our laboratory separated the sloughed-off free cells from the immobilized biomass effectively, thus overcoming wash-out phenomenon frequently encountered in continuous free cell cultures. Furthermore the released-free cells remaining in the bulk solution did not appear to cause substrate mass transfer limitation which was often experienced in suspended mycelial fungal cell fermentations. The primary reason for this was that the suspension broth of the IPRS mainly consisted of roundshaped short mycelial fragments and conidiospores, still remaining Newtonian even at high cell density. In parallel with IPRS bioprocess development, other key factors to be considered necessarily for significant increase in CyA productivity would be strain improvement and medium optimization for the immobilized cells.