• Journal of Korean Society of Water and Wastewater
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• v.23 no.2
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• pp.175-180
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• 2009

This study was conducted to improve biological degradation efficiency of toluene as a model volatile organic compound (VOC) using yeast Candida tropicalis and to suggest an effective method for bioreactor operation. The yeast strain was immobilized with polyethylene glycol (PEG), alginate, and powdered activated carbon (PAC). The yeast-immobilized polymer media were used as fluidized materials in an airlift bioreactor. Polymer media without PAC were also made and operated in another airlift bioreactor. The two bioreactors showed toluene removal efficiencies ranging 80-96% at loading rates of $10-35 g/m^3-hr$, and the bioreactor containing the polymer media with PAC achieved higher removal efficiency. Protein contents in the liquid phase showed that the bioreactor using the yeast-immobilized polymer media with PAC had a higher rate of microbial growth initially than that without PAC. In addition, the microbial growth rate inside of the polymer media with PAC was five times higher than that without PAC. Consequently, the polymer media containing the yeast strain and PAC could enhance removal efficiencies for VOCs, and the immobilization method improve microbial activity and stability for a long-term operation of biological systems.

• 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.

• Journal of Microbiology
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• v.43 no.3
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• pp.301-307
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• 2005

The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, $Cu^{2+}$ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.365-371
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• 1989

Microbiological conversion of sterols to 17-ketosteroids has been recognized as a source for commercial preparation of steroidal drugs. In order to develop bacterial strains and process with Brevibacterium lipolyticum IAM 1398 capable of converting cholesterol to 1,4-Androstadiene-3,17-dione (ADD) at about 27% yield, we studied on strain improvement, fermentation condition and whole cell immobilization. By using UV and/or NTG as mutagens, a mutant to convert cholesterol to ADD with higher yield than 60% was selected. Better production of ADD was manifested in the case of maltose used as a supplemental carbon source, and yeast extract or soytone as a nitrogen source. Addition of tween 80 (0.05%) as a surfactant beneficial for increasing the productivity. The optimal initial pH of the medium was 6.5 and optimal culture temperature was $30^{\circ}C$. Whole cell immobilization by using carrageenan, agar, alginate and acrylamide was carried out and the activity of conversion was tested. In the case of carrageenan and agar, immobilized cells were active for at least two cycles of fermentation.

• KSBB Journal
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• v.15 no.5
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• pp.438-443
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• 2000

Continuous processes using immobilized yeast were investigated in order to shorten beer maturation time. Three silica-based ceramic media and one cellulose-based medium were used. Diacetyl (DA) was one of the most distinctive compounds causing immature flavors. Heat treatment of green beer (GB) to convert a-acetolactate to DA was essential to shorten the time for beer maturation. The longer heat treatment time was needed at the lower temperature. Oxygen concentration in GB had a large influence on the conversion of a-acetolactate to DA. The lower the oxygen concentration in GB, the lower conversion ratio to DA. Heat treated GB was fed continuously to four kinds of immobilized yeast columns. DA concentration after immobilization columns was reduced to less than 0.1ppm at $3∼5^{\circ}C$ 180∼150 minutes retention time in all columns tested. This concentration is enough to fit the quality speification of commercialized product. Formation of a-acetolactate from residual sugars was higher in ceramic media column than cellulose media cloumn. The taste of beers from test processes were not the same as that of traditionally produced beer, but no off-flavors were detected in test samples, which shows that immobilized yeast columns have potentials as rapid processes for beer maturation.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.390-397
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• 1991

Ethanol production by calcium alginate-immobilized baker's yeast (Saccharor/tyces cereviszae) was studied in the batch fermentation using glucose medium as a feed. Immobilied cells were stable between $30^{\circ}C$ and $40^{\circ}C$ whereas free cells were stable between $30^{\circ}C$ and $37^{\circ}C$ The beads were showed constant ethanol productivity during 720 hours (30 days) over. Fermentation characteristics of immobilized baker's yeast were examined changing the initial glucose concentration of broth in fermentation. Initial glucose concentrations employed were 50, 100, 150 and 200 g/l, respectively. In 15% gucose medium, maximum specific growth rate, maximum ethanol yield and ethanol concentration were observed as 0.092 $h^{-1}$, 0.45, 67.5 g/l, respectively.

• Journal of Microbiology and Biotechnology
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• v.7 no.1
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• pp.62-67
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• 1997

To enhance the hyperproductive and low energy-consuming ethanol fermentation rate, the thermotolerant yeast S. cerevisiae RA-74-2 cells were immobilized. An efficient immobilization condition was proved to be $1.5{\%}$ (w/v) alginate solution, neutral pH and 20 h activation of beads. The fermentation characteristics and stability at various temperatures were examined as compared with free S. cerevisiae RA-74-2 cells. The immobilized cells had excellent fermentation rate at the range of pH 3-7 at 30-$42^{\circ}C$ in 15-$20{\%}$ glucose media. When the seed volume was adjusted to 0.12 (v/v) (6ml bead/50 ml medium), $11{\%}$ (w/v) ethanol was produced during the first 34 hand $12.15{\%}$ (w/v) ethanol [$95{\%}$ (w/v) of theoretical yield] during the first 60 h in $25{\%}$ glucose medium. In repetitive fermentation using a 2 litre fermentor, 5.79-$7.27{\%}$ (w/v) ethanol [76-$95{\%}$ (w/v) of theoretical yield] was produced during the 40-55 h in $15{\%}$ glucose media. These data suggested the fact that alginate beads of thermotolerant S. cerevisiae RA-74-2 cells would contribute to economic and hyperproductive ethanol fermentation at high temperature.

• Food Science and Preservation
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• v.6 no.2
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• pp.221-227
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• 1999

The immobilized culture system of Saccharomyces cerevisiae was examined to improve the efficiency of vinegar production from persimmon juice. Optimum concentration of Na-alginate for the immobilization was 2%. When the 1eakage of yeast from get beads was checked by turbidity of culture medium with varying concentration of Na-alginate from 1 to 4%, turbidity of culture medium increased from 8 hrs of cultivation with 1% Na-alginate concentration showing optical density of 0.82 at 20 hrs. However, the increase in turbidity of culture medium was slow with 2-4% Na-alginate showing optical density of 0.55-0.58 at 20 hrs. Microscopical analysis of gel matrix showed that the immobilized yeast was grown well regardless of Na-alginate concentration. Optimum size of gel bead and amount of inoculation were 2-3 m and 33mg, respectively. For ethanol production aerobic cultivation for 121hrs using cohen plug followed by anaerobic cultivation using silicon plug equipped with a check valve was the most effective.

• 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 Korean Society of Environmental Engineers
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• v.31 no.8
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• pp.603-610
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• 2009

This research was performed to improve removal efficiency of toluene and methyl ethyl ketone (MEK) using Candida tropicalis, one of the yeast species. An airlift loop bioreactor (ALB) was employed to enhance the capability of mass transfer for toluene and MEK from the gas phase to the liquid, microbial phase. Polymer gel media made from PAC, alginate and PEG was applied for the effective immobilization of the yeast strain on the polymer gel media. The experimental results indicated that the mass transfer coefficient of toluene without polymer gel media was 1.29 $min^{-1}$ at a gas retention time of 15 sec, whereas the KLa value for toluene was increased to 4.07 $min^{-1}$ by adding the media, confirming the enhanced mass transfer of volatile organic compounds between the gas and liquid phases. The removal efficiency of toluene and MEK by using yeast-immobilized polymer gel media in the ALB was greater than 80% at different pollutant loading rates (5, 10, 19 and 37 g/$m^3$/hr for toluene, 4.5, 8.9, 17.8 and 35.1 g/$m^3$/hr for MEK). In addition, an elimination capacity test conducted by changing inlet loading rates stepwise demonstrated that maximum elimination capacities for toluene and MEK were 70.4 and 56.4 g/$m^3$/hr, respectively.