• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.31-36
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• 1991

Yeast expression plasmids containing an appropriate leader sequence and bovine $\beta$-casein cDNA were constructed to produce $\beta$-casein for the study of its functional characteristics. Two kinds of expression systems for $\beta$-casein were constructed using pCGYl444 as a precursor plasmid. This plasmid is a yeast-E. coli shuttle vector which contains the chelatin promoter. The plasmid pISB202 contains the invertase leader sequence and $\beta$-casein gene. The plasmid pDEB303 contains the original bovine $\beta$-casein leader sequence gene. These two plasmids were introduced into S. cerevisiae AB116 which is a strain deficient in the major yeast proteinases. Each clone was grown in minimal media for 24 h before induction by $CuSO_4$. The cells were thus grown under expression conditions. Both strains harbouring pISB202 and pDEB303 expressed bovine $\beta$-casein. The $\beta$-casein was detected using immunochemical staining after western blot. Secretion of $\beta$-casein was detected in the culture broth. The estimated amount of secreted $\beta$-casein was approximately 50 ${\MU}g$/l.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1161-1168
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• 2009

Ethanol production by the simultaneous saccharification and fermentation (SSF) of low-value rice wine cake (RWC) without cooking was investigated. RWC is the filtered solid waste of fermented rice wine mash and contains 53% raw starch. For the SSF, the RWC slurry was mixed with the raw-starch-digesting enzyme of Rhizopus sp. and yeast, where the yeast strain was selected from 300 strains and identified as Saccharomyces cerevisiae KV25. The highest efficiency (94%) of ethanol production was achieved when the uncooked RWC slurry contained 23.03% starch. The optimal SSF conditions were determined as 1.125 units of the raw-starch-digesting enzyme per gram of RWC, a fermentation temperature of $30^{\circ}C$, slurry pH of 4.5, 36-h-old seeding culture, initial yeast cell number of $2{\times}10^7$ per ml of slurry, 17 mM of urea as the nitrogen additive, 0.25 mM of $Cu^{2+}$ as the metal ion additive, and a fermentation time of 90 h. Under these optimal conditions, the ethanol production resulting from the SSF of the uncooked RWC slurry was improved to 16.8% (v/v) from 15.1% (v/v) of pre-optimization.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.287-290
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• 2000

To examine effects of agitation and aeration as well as adding of glucose and yeast extract on cell growth and polysaccharide production by Paecilomyces japonica, batch culture was carried out at 5L jar fermenter at $27^{\circ}C$ with the initial pH 7 for 7 days cultivation(innoculum size 2%, working volume 3L). Media compositions(g/L) were 30 glucose, 20 yeast extract, 0.5 $KH_2PO_4$, $0.1\;CuCl_2\;{\cdot}\;2H_2O$. Optimum culture conditions of agitation and aeration in batch culture were 400 rpm and 1.0 vvm, resulting in 23.1 g/L biomass and 2.5 g/L polysaccharide. Additional feeding of glucose and yeast extract with a pulse mode conferred an advantage on cell growth and polysaccharide production with showing the results of 29.2 g/L and 3.3 g/L, respectively.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.566-571
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• 2005

A natural habitat bacterium, Enterobacter cloaceae K41 was isolated from fresh water plant root and identified. This strain was used to investigate heavy metal resistance. The optimal growth conditions of the bacterium were LB medium containing$1\%$ yeast extract, $1\%$ lactose, $1\%$ NaCl, pH 7.0, at $37^{\circ}C$, and for 24 hours on a shaker. The minimal inhibitory concentration (MIC) of heavy metals against E. cloaceae KCTC2519 and E. cloaceae K41 was compared. The MIC of E. cloaceae K41 was 150 ppm in Cu, 50 ppm in Cd whereas that of the standard strain was 50 ppm in Cu but no growth was observed either Cd or two mixed heavy metal solution. The presence of plasmid was cleared from the isolated strain whereas no possession from the standard strain. The plasmid from E. cloaceae K41 was transformed into E. coli $DH5{\alpha}$. The MIC of transformed strain increased resistance 7 times in Cu and 6 times in Cd by insertion of this plasmid. The metal adsorption of the transformant was increased 1.3 times in Cu and 1.5 times in Cd indicating the plasmid was responsible for heavy metal resistance.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.5
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• pp.631-636
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• 1998

Candida lipolytica(C. lipolytica) FM5 was selected as one of the strong saprophytic yeasts isolated from mackerel (Scomber japonicus). The selected strain could produce extracellular proteolytic enzyme. The effective medium for production of proteolytic enzyme by C. lipolytica FM5 was TPPY broth containing Bacto-tryptone $0.5\%$, proteose peptone $0.5\%$, yeast extract $0.25\%$, NaCl $0.5\%$ and $CaCl_2\;0.2\%$. The pattern of proteolytic enzyme production by C. lipolytica FM5 was the almost same as that of growth curve of the strain. Namely, the enzyme production was begun from the early stage of exponential phase and it was reached the highest at the begining of the stationary phase of the yeast growth. The optimum toeperature of the produced proteolytic enzyme was $35^{\circ}C$ and its activity was not significantly changed by pH between 6.5$\~$9.0 and also it was not significantly affected by several kinds of cations such as $Ca^{2+},\;Cu^{2+},\;Fe^{2+}$ and $Mg^{2+}$ but it was affected negatively by some cations such as $Zn^{2+},\;Mn^{2+}$ and $K^+$.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.69-74
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• 2000

The medium for erythritol production by Torula sp. in a 500-ml baffled flask was optimized to be 300 g/I sucrose, 10 g/I yeast extract, 3 g/I $KH_2PO_4$, and 10 mg/I $CuSO_4{\cdot}5H_2O{\;}at{\;}34^{\circ}C$ with initial pH of 5.5. Using this optimal medium, erythritol of 166 g/I was obtained after 140 h of cultivation, corresponding to 55.3% of the erythritol yield from sucrose with a productivity of 1.11 g/I/h. Optimal concentrations of carbbon and nitrogen sources in a fermentor were higher than that in a flask due to the higher oxygen supply of the fermentor. Employing the medium containing 300 g/I or 400 g/I sucrose for the determination of optimal C/N ratio, the C/N ratio was found to be more important than the nitrogen concentration for effective erythritol production, The optimal ratio of yeast extract to sucrose (g/g) was 20. The yield and productivity of erythritol were maximal in the medium containing 400 g/I sucrose and 20 g/I yeast extract. when dissolved oxygen in the culture was increased, the cell mass increased but the erythritol production was manimal in the range of 5 to 10% of dissolved oxygen. Under the optimal the rane of 5 to 10% of dissolved oxygen. Under the optimal culture condition of the fermentor, a final erythritol concentration of 200 gI was obtained after 120 h with a yield of 50% and the productivity was 1.67 g/I/h. The yield was the highest among erythritol-producting microorganisms

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.359-363
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• 1986

S. diastaticus hydrolysised $\alpha$-1.4 linkage of the starch and was known fermenting yeast strain, but poorly hydrolized $\alpha$-1.6 linkage of the starch. To improve the starch fermentation ability of yeast, we tried that protoplast fusion between S. diastaticus and C. tropicalis and finally two starins of fusant (FPDC42, FPDC43) were obtained. C. tropicalis well hydrolysis both $\alpha$-1.4 and $\alpha$-1.6 linkanges in the starch. The protoplast of parental auxotrophic cells were fused in the presence of 10mM CaCl$_2$ and 35% of polyethylene glycol (M. W. 4,000). The fusion frequency was 10$^{-5}$ to 10$^{-6}$. Properties of the fusants(genetic stability, assimilation of carbon sources, random spore formation, copper resistance, NaCl tolerance, DNA content, cell size and growth rate) were investigated.

• Mycobiology
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• v.49 no.5
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• pp.507-520
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• 2021

Papiliotrema huenov was previously reported to be highly tolerant of a range of extremely toxic heavy metals. This study aimed to identify the potential of P. huenov to remove manganese and copper from aqueous solution. Physical conditions which affect removal of Mn(II) and Cu(II) were determined. Optimal temperature for adsorption of both metal ions was 30 ℃, and optimal pH for maximum uptake of Mn(II) and Cu(II) were 5 and 6, respectively. Under these conditions, living cells of P. huenov accumulated up to 75.58% of 110 mg/L Mn(II) and 70.5% of 128 mg/L Cu(II) over 120 h, whereas, the removal efficiency of metal ions by dead cells over 1 h was 60.3% and 56.5%, respectively. These results indicate that living cells are more effective than dead biomass for bioremediation, but that greater time is required. The experimental data extends the potential use of P. huenov in biosorption and bioaccumulation of toxic heavy metals to copper and manganese, two of the most common industrial contaminants.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.331-336
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• 2007

A bacterium producing the alkaline pretense was isolated from Chungkookjoug, and was identified as Bacillus subtilis JK-1 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The optimum pH and temperature of the pretense activity were pH 9.0 and $55^{\circ}C$, respectively. This enzyme was stable at the temperatures $40{\sim}80^{\circ}C$. The maximum alkaline pretense production was obtained when 1.0% (w/v) xylose, 1.0% (w/v) yeast extract and 0.3% (w/v) $CuSO_4$ were used as carbon source, nitrogen source and mineral source. Under the optimal condition, growth of the isolate was reached at stationary phase after 12 hr followed by incubation, the alkaline pretense production reached a maximum level with $16{\sim}20$ hr cultivation.

• The Korean Journal of Mycology
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• v.49 no.3
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• pp.337-349
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• 2021

This study aimed to isolate wild yeasts from water and soil sample of the Nonsan stream and Sapgyoho (lake) in Chungcheonnam-do, Korea, and to further characterize previously unrecorded wild yeast strains. In total, 102 strains, representing 55 different species of wild yeast were isolated from 95 samples collected from the Jangseoncheon and Ipchoncheon of Nonsan stream in Jellabuk-do and Chungcheonnam-do. Among these, 33 strains were isolated from alkalophilic yeast extract-peptone-dextrose (YPD) medium (pH 9.0), and 9 strains were isolated concurrently on general YPD medium (pH 6.5) and alkalophilic medium. Seventeen strains of Cryptococcus laurentii were predominantly isolated. Additionally, 65 strains, representing 27 different species of wild yeast were isolated from 58 samples obtained from Sapgyoho (lake). Among the 82 isolated wild yeast strains, 8 strains, including Candida fructus JSC 72-1(JSL-GGU 015), had not previously been recorded. All 8 previously unrecorded yeasts were oval in shape except C. fructus JSC72-1(JSL-GGU-015), and only the Filobasidium chernovii JSC39-1(JSL-GGU-013) strain formed spores. All strains except Pseudosydowia eucalypti JSC23-6(JSL-GGU-012) grew well in yeast extract-peptone-dextrose (YPD) and yeast extract-malt extract media and grew in vitamin-free medium. Four strains, including P.eucalypti JSC23-6(JSL-GGU-012) grew well in 15% NaCl-containing YPD medium. F.chernovii JSC39-1(JSL-GGU-013) and Sirobasidium intermedium JSC7-3(JSL-GGU-014) assimilated lactose, and five strains, including F. chernovii JSC39-1(JSL-GGU-013) also assimilated starch. All strains were resistant to 800 ppm of Ca, Cu, Li, and Mg ions.