• KSBB Journal
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• v.11 no.1
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• pp.30-36
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• 1996

Based on fermentation data for cyclosporin A production, simple Monod kinetics was proposed for both immobilized and suspended cultures. Higher value of $\mu$mas and lower value of Km suggest better catalytic activity of the immobilized cells than the parallel suspended cells. Furthermore, lower Km value in the immobilized cell system indicates higher affinity of the immobilized cells for carbon substrate as compared with the suspended cells. For immobilized cell cultures, these parameters were also utilized for the estimation of effectiveness factor, an indicator for intraparticle mass transfer resistance. Based on simulation studies, optimum radius of celite beads was turned out $100 ~ 500{\mu}m$In this simulation work, we examined the influence of biosupport size and immobilized biomass density on diffusional resistance of substrate inside the bead matrix. In order to maintain uniformly distributed cell activities in biosupport, it was essential to determine optimum slze of particle, and then to estimate the most economic loaded biomass content.

• BMB Reports
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• v.32 no.1
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• pp.98-102
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• 1999

Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

• KSBB Journal
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• v.5 no.2
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• pp.167-173
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• 1990

This study is to investigate for obtaining the operating conditions of continuous vinegar production using fluidized bed reactor by Acetobacter aceti cell immobilized in Ca-alginate gel. The optimum conditions obtaining by batch fermentation using fluidized bed reactor were as follows; The fermentation temperature and aeration rate were 3$0^{\circ}C$ and 1.0VVM and the initial concentration of ethanol and acetic acid in medium were 33g/l and 27g/l respectively. The amount of bead used was 25%(w/v). The overall acetic acid productivities of batch fermentations by free cell and immobilized cell were 0.31g/l-hr and 0.48g/l-hr, respectively, at the final acetic acid concentration of 50g/l. In the continuous vinegar production using fluidized bed reactor by immobilized cell under optimum conditions, it was possible to produce 23g/l acetic acid continuously up to 90 days with maximum acetic acid productivity of 2.76g/l-hr at dilution rate 0.12hr-1.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.268-268
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• 2006

Due to their multipotency, stem cells can differentiate into a variety of specialized cell types, such as chondrocytes, osteoblasts, myoblasts, and nerve cells. As an alternative to mature tissue cells, stem cells are of importance in tissue engineering and regenerative medicine. Since interactions between scaffold and cells play an important role in the tissue development in vitro, synthetic oligopeptides have been immobilized onto polymeric scaffolds to improve specific cell attachment and even to stimulate cell differentiation. In this study, chondrogenic differentiation of stem cells was evaluated using surface-modified PLLA scaffolds, i.e., either hydrophilic acrylic acid (AA)-grafted PLLA or RGD-immobilized one. Porous PLLA scaffolds were prepared using a gas foaming method, followed by plasma treatment and subsequent grafting of AA to introduce a hydrophilicity (PLLA-PAA). This was further processed to fix RGD peptide to make an RGD-immobilized scaffold (PLLA-PAA-RGD). Stem cells were seeded at $1{\times}10^{6}$ cells per scaffold and the cell-PLLA constructs were cultured for up to 4 weeks in the chondrogenic medium. Using these surface-modified scaffolds, adhesion, proliferation, and chondrogenic differentiation of stem cells were evaluated. The surface of PLLA scaffolds turned hydrophilic (water contact angle, 45 degrees) with both plasma treatment and AA grafting. The hydrophilicity of RGD-immobilized surface was not significantly altered. Cell proliferation rate on the either PLLA-PAA or PLLA-PAA-RGD surface was obviously improved, especially with the RGD-immobilized one as compared to the control PLLA one. Chondrogenic differentiation was clearly identified with Safranin O staining of GAG in the AA- or RGD-grafted PLLA substrates. This study demonstrated that modified polymer surfaces may provide better environment for chondrogenesis of stem cells.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.1
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• pp.19-22
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• 2003

The semicontinuous production of red pigment by immobilized cells of Bacillus sp. B H-99 was investigated in comparison with free cells. The red pigment produced highest productivity under the conditions of aeration of 0.2 mL/min and 2 mm diameter of gel beads by using 3.0% sodium alginate. Semicontinuous production by immobilized cells showed the highest productivity with replacement of fresh production medium in every 72 h for fourth fermentation cycle following the conditions of red pigment productivity.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.130-135
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• 1991

The effects of media composition on citric acid fermentation using surface immobilized Saccharomycopsis lipolytica were studied. The use of the standard medium for these organisms resulted in rapid decrease of citric acid production and a transformation of immobilized cell morphologies from a yeast-type to a mycelium-type. When the standard medium was enriched with vitamins, trace minerals, a growth factor and ammonium to form a Vigorous Stationary Phase (VSP) fermentation type medium, relatively stable citric acid production (10 mg/lㆍh) was obtained. Using the VSP type medium, the surface immobilized cells also retained their yeast-type form.

• The Korean Journal of Food And Nutrition
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• v.11 no.3
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• pp.319-322
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• 1998

The whole cell of alkalophilic Streptomyces sp. B-2 which produce glucose isomerase was immobilized by entrapment method for the effective production of high fructose syrup. The highest immobilized activity was achieved when the enzyme was bound to 2% $textsc{k}$-carrageenan. Immobilized glucose isomerase the pH optimum was about pH 7.5~8.5. Immobilization of alkalophilic Streptomyces sp. B-2 on 2% $textsc{k}$-carrageenan at 7$0^{\circ}C$ showed an increase in glucose isomerase activity. GI activity of immobilized cells was maximum Co2+ concentration 10-3M, Mg2+ concentration 10-3M.

• Journal of Microbiology and Biotechnology
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• v.5 no.4
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• pp.229-233
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• 1995

Acremonium chrysogenum was immobilized in ionotropic gel beads to develop semi-continuous production of cephalosporin C (CPC). Barium alginate beads were more stable than calcium alginate or strontium alginate beads in chemically defined media. The gel stability of Ba-alginate was further increased by cross-linking with polyethyleneimine (PEI). The presence of carboxymethyl cellulose inside Ba-alginate beads did not reduce mass transfer resistance. Ba-alginate microbeads that had little diffusion limitation increased CPC production rate 1.6 fold higher than that of normal beads. CPC fermentation with immobilized cells in Ba-alginate microbeads was performed continuously for 40 days by way of repeated fed-batch operations. Mathematical modeling was developed to describe the repeated fed-batch fermentation system. Results of the computer simulation agreed well with the experimental data, which made it possible to predict an optimal feeding rate that could maximize total CPC productions.

• KSBB Journal
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• v.30 no.2
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• pp.82-90
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• 2015

Plant cell immobilization can protect plant cells from shear forces and increase the stability of gene. An additional advantage of immobilization is the easiness for performing continuous culture with cell recycling. Therefore plant cell immobilization can overcome the limitations of plant cell applications. In addition, target protein should be selected from pharmaceutical proteins to get rid of low expression level problem. The enhanced production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in immobilized Nicotiana tabacum suspension cell cultures. When the cells were immobilized in polyurethane foam, specific production of hGM-CSF was higher than that in alginate bead immobilization. Optimum continuous culture condition was the addition of 60 g/L sucrose in growth media with exchanging media every 6 day. Under the same condition, specific hGM-CSF production was 7 times higher in a 500-mL spinner flask than that in 100-mL Erlenmeyer flasks. Therefore, development of an effective immobilization process would be possible when the advantage of easy cell recycling was used. Consequently, enhanced production of target proteins could be possible in immobilized continuous cultures when the advantages of immobilization were applied.