• Journal of Microbiology and Biotechnology
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• v.20 no.9
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• pp.1314-1321
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• 2010

Sulfur metabolism in S. cerevisiae is well established, but the mechanisms underlying the formation of sulfide remain obscure. Here, we investigated by real-time RT-PCR the dependence of expression levels of MET3, MET5/ECM17, MET10, MET16, and MET17 along with SSU1 on nitrogen availability in two wine yeast strains that produce divergent sulfide profiles. MET3 was the most highly expressed of the genes studied in strain PYCC4072, and SSU1 in strain UCD522. The strains behaved differently according to the sampling times, with UCD522 and PYCC4072 showing the highest expression levels at 120 h and 72 h, respectively. In the presence of 267 mg assimilable N/l, the genes were more highly expressed in strain UCD522 than in PYCC4072. MET5/ECM17 and MET17 were only weakly expressed in both strains under any condition tested. MET10 and SSU1 in both strains, but MET16 only in PYCC4072, were consistently upregulated when sulfide production was inhibited. This study illustrates that strain genotype could be important in determining enzyme activities and therefore the rate of sulfide liberation. This linkage, for some yeast strains, of sulfide production to expression levels of genes associated with sulfate assimilation and sulfur amino acid biosynthesis could be relevant for defining new strategies for the genetic improvement of wine yeasts.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1539-1545
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• 2010

In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.184-189
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• 2012

We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.923-927
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• 2005

ln order to develop yeast cells that produce a bacteriocin on their cell surfaces, the 114 bp Leucocin A gene with stop codon was ligated into pYDl, an yeast vector. The recombinant DNA, pYDl-LeucoA was used to transform yeast (Saccharomyces cerevisiae) cells. Yeast cells harboring pYDl-LeucoA showed antibacterial activity against Bacillus subtilis. To confirm these bacteriocidal yeast cells possess the Leucocin A gone, PCR was performed with plasmid prepared from transformed yeast cells as a template and two Leucocin A-specific primers. In this study, bacteriocidal yeast cells that can be used as an antibiotic or a food preservative were developed.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.633-643
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• 2017

To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.

• Korean Journal of Food Science and Technology
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• v.9 no.2
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• pp.97-105
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• 1977

1) In order to obtain a microbial strain having a strong yeast cell wall lytic activity, about 156 isolates capable of forming clear zones on baker's yeast-peptone-bouillon agar plate were obtained from soil, mud and water samples and a strain K-42 with the highest lytic activity was identified as Bacillus circulans. 2) Effect of carbon sources on the lytic enzyme production by the K-42 strain was in the decreasing order of maltose>glucan>xylose>control in 2-day culture and of lactose>galactose>glucan>control in 3-day culture. Effect of inorganic nitrogen sources was in the decreasing order of ammonium acetate>sodium nitrate>control in 2-day culture and of ammonium chloride>ammonium oxalate>control in 3-day culture, whereas organic nitrogen sources except milk casein showed an increase in 2-day culture and a decrease in 3-day culture. Synergistic effect of carbon sources and nitrogen sources was not observed. 3) The enzyme production by the K-42 strain was greatly affected by pH change of the culture medium, thus a high lytic activity could be maintained by keeping the pH range of $7{\sim}8$ and adding carbon or nitrogen sources. 4) Optimum conditions for the lytic activity of the K-42 strain were obtained at $pH\;7{\sim}8$ and $60^{\circ}C$ and the extent of hydrolysis toward heated yeast cell wall was 65%.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.373-381
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• 2022

Strain E4, an unrecorded species of dimorphic fungi, was isolated from the gut of earthworms collected in Gyeonggi Province, South Korea. Nucleotide sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region revealed that this species is a member of the genus Blastobotrys, Blastobotrys illinoisensis. Strain E4 differed from its closest known species, B. mokoenaii and B. malaysiensis, by harboring 3-5 and 12-14 nucleotide substitutions in the D1/D2 and ITS regions, respectively. Phylogenetic analysis based on concatenated sequences of the D1/D2 region of the LSU rRNA gene and the ITS region also indicated that strain E4 belongs to the Blastobotrys clade and is distinct from other related species in the clade. The previously unreported isolate could be distinguished from closely related species by its inability to ferment carbon sources. To our knowledge, this is the first report on the isolation of Blastobotrys species from the gut of earthworms in Korea. The strain used was E4 (=KCTC 27831=JCM 33428).

• The Korean Journal of Mycology
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• v.46 no.3
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• pp.307-314
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• 2018

The goal of this study was to investigate the microbiological characteristics of the ethanol-producing wild yeast, Aureobasidium pullulans P-1, isolated from flowers growing near the Yedang reservoir, Chungnam province, Korea, and in addition, to optimize its fermentation ability for the production of Makgeolli. A. pullulans P-1 was oval-shaped, and formed ascospores and pseudomycelium. The P-1 strain was a halophilic and sugar tolerant yeast which grew in 15% NaCl and 50% glucose-containing yeast extract-peptone-dextrose media. The P-1 strain was also resistant to 20% ethanol. Changes of the physicochemical properties during Makgeolli fermentation by A. pullulans P-1 were investigated. A maximum of 8.45% ethanol was obtained when a mixture of cooked rice, 150% water, and 35% ipguk per cooked rice was fermented by 5% A. pullulans P-1 culture broth at $25^{\circ}C$ for 10 days. Antihypertensive angiotensin I-converting enzyme inhibitory activity in the Makgeolli ferment produced by A. pullulans P-1 reached a high of 71.1% after 10 days.

• The Korean Journal of Mycology
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• v.48 no.3
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• pp.229-235
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• 2020

The goal of this study was to isolate and identify the wild yeast strains in intestine of Othius punctulatus collected in Cheongju-si, Chungcheongbuk-do, Korea and characterize their unrecored yeast strains. We isolated 16 wild yeast strains from 5 intestine samples from Othius punctulatus. The 16 wild yeast strains were included three strains in the genus Aureobasidium, four strains in the genus Cystofilobasidium, one strain in the genus Mrakia, three strains in the genus Naganishia, two strains in the genus Saitozyma, two strains in the genus Sampaiozyma and one strain in the genus Scheffersomyces. Among them Cystofilobasidium capitatum YP53, Cystofilobasidium capitatum YP71, Cystofilobasidium macerans YS620, Cystofilobasidium macerans YS622, Mrakia aquatica YP158 and Scheffersomyces stipitis YI5 were recorded for the first time in Korea. The microbiological characteristics and carbon assimilation of these previously unrecorded yeasts were investigated. Almost all of them were oval-shaped. And, glycerol, L-arabinose, xylitol, and inositol are not utilized.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.202-206
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• 2000

Feasibility of microbial culture media using by-product of salt fermented kanary was investigated. Gram negative strain, Escherichia coli, and Gram positive strain, Bacillus subtilis, and bioluminescent Photobacterium Phosphoreum were incubated with kanary by-Product media (KB media). Compared with LB media, KB media had enough carbon source, but lacked nitrogen source and growth factor. When 0.5% of peptone as a nitrogen source and 0.3% of yeast extract as nitrogen and growth factor source were fortified in KB media, the cell population rate was similar to LB media. Also, when 0.5% of yeast extract was fortified to KB media, it showed the same result as in LB media. The price of KB media with fortification of 0.5% peptone and 0.3% yeast extract, and 0.5% of yeast extract is only 46 and 19% of that of LB media, respectively. These results showed that kanary by-Product could be a good and cheaper bacterial culture media if small amount of nitrogen source and growth factor were added.