• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.717-725
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• 1996

We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

• Clean Technology
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• v.4 no.1
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• pp.45-59
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• 1998

Continuous immobilized-cell culture was carried out for the production of kasugamycin, a secondary metabolite by a filamentous bacteria, Streptomyces kasugaensis, with an intention of reducing waste generation. A sporulation medium was developed for production of bulk amounts of spores, and the spores were entrapped into celite biosupports for immobilization. It was possible to effectively keep the immobilized-cells inside the reactor during the continuous culture by an efficient immobilized cell separator of decantor type on the outlet of the fermentor. Using this continuous immobilized-cell fermentor system, we investigated the effects of feed substrate and phosphate concentrations on kasugamycin production and chemical oxygen demand(COD). Comparing with the conventional suspended-cell batch culture, the kasugamycin productivity was observed to increase by 2.5 times, whereas COD per unit kasugamycin production decreased by 2.3 times in the continuous immobilized-cell culture. Based on these results, the continuous immobilized-cell system was considered to be a cleaner bioprocess than the conventional batch suspended-cell system.

• 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.