• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.468-472
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• 2003

Recently, genetic engineering techniques have been used to display various heterologous peptides and proteins (enzyme, antibody, antigen, receptor and fluorescence protein, etc.) on the yeast cell surface. Living cells displaying various enzymes on their surface could be used repeatedly as 'whole cell biocatalysts' like immobilized enzymes. We constructed a yeast based whole cell biocatalyst displaying T. reesei cellobiohydrolase I (CBH I ) on the cell surface and endowed the yeast-cells with the ability to degrade cellulose. By using a cell surface engineering system based on ${\alpha}-agglutinin,$ CBH I was displayed on the cell surface as a fusion protein containing the N-terminal leader peptide encoding a Gly-Ser linker and the $Xpress^{TM}$ epitope. Localization of the fusion protein on the cell surface was confirmed by confocal microscopy. In this study, we report on the genetic immobilization of T. reesei CBH I on the S. cerevisiae and hydrolytic activity of cell surface displayed CBH I.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.165-169
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• 1989

To achieve higher ethanol productivity in the fermentation system, a continuous ethanol production has been investigated with the air-supplement in a packed-bed immobilized cell reactor system. Yeast cells were immobilized using sodium alginate gel. The results showed that, when the feed medium was saturated with oxygen through aeration into the medium reservoir, the maximum ethanol productivity of the reactor was enhanced from 35 g/$\ell$-gel-hr to 55 g/$\ell$-gel-hr at the residence time of 10-20 min. and the residence time for the 90% conversion of substrate to ethanol was reduced from 40 min. to 25 min. In case of 18% glucose medium, the maximum productivity was increased from 35 g/$\ell$-gel-hr to 45 g/$\ell$-gel-hr and time required for 90% conversion was from 90 min to 70 min. This behavior of air-supplemented reactor system might be due to the fact that both growth and viable fraction of yeast within the Eel were increased during reactor operation.

• Korean Journal of Food Science and Technology
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• v.25 no.2
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• pp.154-159
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• 1993

This present study was carried out particulary focusing on rapid fermentation of soy sauce by using column type reactor $(30\;cm{\times}5\;cm)$ packed each immobilized cells of Pediococcus halophilus R-22, Saccharomyces rouxii R-60 and Candida etchellsii H-50. When immobilized P. halophilus R-22 by column type reactor was performed continuously fermentation, lactic acid was produced $0.62{\sim}0.64%$ during 25 days and then decreased gradually after 30 days. S. rouxii R-60 was Produced the $2.1{\sim}2.5%$ ethylalcohol constantly for 35 days and also C. etchellsii H-50 was produced $14{\sim}16\;mg/l$ 4-ethylguaiacol for 35 days and then this products were decreased gradually after fermentation of 40 days. Final Products of fish sauce contained 1,721.6 mg% total nitrogen, 1,584.1 mg% amino-nitrogen, 0.75% lactic acid, 2.7% ethylalcohol and 18.2 mg/l 4-ethylguaiacol.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.373-380
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• 1993

When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.511-517
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• 2013

In this work, a fibrous bed bioreactor with high specific surface area and good adsorption efficacy for S. cerevisiae cells was used as the immobilization matrix in the production of ethanol. In batch fermentation, an optimal ethanol concentration of 91.36 g/l and productivity of 4.57 g $l^{-1}\;h^{-1}$ were obtained at an initial sugar concentration of 200 g/l. The ethanol productivity achieved by the immobilized cells was 41.93% higher than that obtained from free cells. Ethanol production in a 22-cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in packed-bed reactors, a maximum ethanol concentration of 108.14 g/l and a productivity of 14.71 g $l^{-1}\;h^{-1}$ were attained at $35^{\circ}C$, and a dilution rate of 0.136 $h^{-1}$ with 250 g/l glucose.

• Applied Biological Chemistry
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• v.39 no.1
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• pp.1-8
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• 1996

A one-stage, continuous-flow bioreactor with both immobilized and suspended cells was used to investigate the roles of lipid supplements in ethanol production by Saccharomyces cerevisiae. The reactor performance and the level of alcohol dehydrogenase(ADH) activities of the suspended cells, grown under various conditions, were measured. When ergosterol and/or oleic acid were added with surfactants to the yeast culture grown under non-aerated conditions, remarkable increases in ethanol production and cell growth was achieved, but specific ADH activities were not affected. Especially, no difference of specific ADH activities of the suspended cells grown under aerated and non-aerated condition was observed. The addition of the surfactant as a supplement also resulted in significant increases in ethanol production, cell growth, and specific ADH activity. When ergosterol and oleic acid were added to the yeast culture exposed to higher ethanol concentration($>40\;g/{\ell}$) level, ethanol production, cell growth, and specific ADH activity were increased, but the addition of surfactant was as effective as at lower ethanol concentration level. The results indicated that lipid supplements were more effective at higher ethanol concentration level than at lower ethanol concentration level during ethanol production. ADH isozyme patterns of the yeast cultures grown under various conditions on starch gel electrophoresis showed only one major band, probably ADH I. The migrating distance of the major isozyme, however, varied slightly according to the culture conditions of the cells. No apparent correlation was found between specific ADH activity and amount of ethanol produced. Cell mass was more important factor for ethanol production than specific ADH activity of the cells.

• Bulletin of the Korean Chemical Society
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• v.29 no.2
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• pp.381-388
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• 2008

An X-linked skeletal disorder, SEDT (spondyloepiphyseal dysplasia tarda) is a genetic disease characterized by a disproportionately short trunk and short stature caused by mutations in the SEDL gene. This gene is evolutionarily conserved from yeast to human. The yeast SEDL protein ortholog, Trs20p, has been isolated as a member of a large multi-protein complex called the transport protein particle (TRAPP), which is involved in endoplasmic reticulum (ER)-to-Golgi transport. The interaction between SEDL and partner proteins is important in order to understand the molecular mechanism of SEDL functions. We isolated several SEDL-binding proteins derived from rat cells by an immobilized GST-fusion method. Furthermore, the SEDL-homologue proteins were identified using motif based methods. Common motifs between SEDL-binding proteins and SEDL-homologue proteins were classified into seven types and 78 common motifs were revealed. Sequence similarities were contracted to seven types using phylogenetic trees. In general, types I-III and VI were classified as having the function of acetyl-CoA carboxylase, glycogen phosphorylase, isocitrate dehydrogenase, and enolase, respectively, and type IV was found to be functionally related to the GST protein. Types V and VII were found to contribute to TRAPP vesicle trafficking.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.1
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• pp.47-52
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• 1989

Aspergillus awamori, B-2 which has a high ability to decolorize melanoidin was selected among various fungi. Aspergillus awamori, B-2 showed the highest decolorization activity when it was cultivated in a melanoidin medium containing 3.0% glucose, 0.5% yeast extract, 0.1 % $KH_2PO_4\;and\;0.05%\;MgSO_4{\cdot}7H_2O$ at an initial pH 7.0 at $37^{\circ}C$ for 5 days. Mycelia immobilized system with. Ca-alginate was more effective on melanoidin decolorization activity showed approximately 70% in 10 days under the optimal conditions. Continuous decolorization of melanoidin using, reuse of immobilized mycelia showed an almost constant decolorization of abort 60-70% for 15 days.

• Journal of Microbiology and Biotechnology
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• v.3 no.1
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• pp.24-30
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• 1993

In order to develop a novel yeast which produces the charateristic aroma of soy sauce, a protoplast fusion between Zygosaccharomyces rouxii WFS4 and Torulopsis versatilis IAM 4993 was carried out. Auxotrophic mutants as selective markers were obtained from Zygosaccharomyces rouxii and Torulopsis versatilis by treatment of N-methyl-N -nitro-N-nitrosoguanidine. The conditions of the protoplast formation and the regeneration for fusion were examined. The protoplast fusion using polyethylene glycol 4000 led to the fusion frequency of $4~5{\times}10^{-7}\;cells/ml$. Among fusants, a fusant ST723-F31 presented the best results in terms of the aromaticity of fragrance, the growth pattern, the resistance against salt and the degree of growth according to pH. It makes easy to control the production and the balance of aroma components so that it gives a good flavor, shortens the fermentation period and, simplifies the preparation process when using a bioreactor into which fusant is immobilized.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.142-149
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• 1991

Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.