• Title/Summary/Keyword: Wild yeast

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The Gene Encoding γ-Glutamyl Transpeptidase II in the Fission Yeast Is Regulated by Oxidative and Metabolic Stress

  • Kang, Hyun-Jung;Kim, Byung-Chul;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • v.38 no.5
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    • pp.609-618
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    • 2005
  • $\gamma$-Glutamyl transpeptidase (GGT, EC 2.3.2.2.) catalyzes the transfer of the $\gamma$-glutamyl moiety from $\gamma$-glutamyl containing ompounds, notably glutathione (GSH), to acceptor amino acids and peptides. A second gene (GGTII) encoding GGT was previously isolated and characterized from the fission yeast Schizosaccharomyces pombe. In the present work, the GGTII-lacZ fusion gene was constructed and used to study the transcriptional regulation of the S. pombe GGTII gene. The synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene was significantly enhanced by NO-generating SNP and hydrogen peroxide in the wild type yeast cells. The GGTII mRNA level was increased in the wild-type S. pombe cells treated with SNP. However, the induction by SNP was abolished in the Pap1-negative S. pombe cells, implying that the induction by SNP of GGTII is mediated by Pap1. Fermentable carbon sources, such as glucose (at low concentrations), lactose and sucrose, as a sole carbon source, enhanced the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in wild type KP1 cells but not in Pap1-negative cells. Glycerol, a non-fermentable carbon source, was also able to induce the synthesis of $\beta$-galactosidase from the fusion gene, but other non-fermentable carbon sources such as acetate and ethanol were not. Transcriptional induction of the GGTII gene by fermentable carbon sources was also confirmed by increased GGTII mRNA levels in the yeast cells grown with them. Nitrogen starvation was also able to induce the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in a Pap1-dependent manner. On the basis of the results, it is concluded that the S. pombe GGTII gene is regulated by oxidative and metabolic stress.

Isolation of Wild Yeasts from Humus-rich Soil in City Park of Daejeon Metropolitan City, Korea, and Characterization of the Unrecorded Wild Yeasts (대전광역시 도시 자연공원 부엽토와 주변 산림토양들로부터 야생효모의 분리, 동정 및 국내 미기록 야생효모들의 특성)

  • Han, Sang-Min;Lee, Sang-Yeop;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.46 no.1
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    • pp.75-82
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    • 2018
  • Totally 91 strains of wild yeasts were isolated from the humus rich soil in the Bogyong city park of Daejeon city, Korea. Majority of the strains belonged to Cryptococcus spp., which included 11 strains of Cryptococcus aureus. Among them, Kwoniella mangroviensis JSS0500, Candida corydalis JSS0501, Candida bombi JSS0503, Candida multigemmis JSS0504, Cryptococcus dimennae JSS0506, Cryptococcus saitoi JSS0507, Cryptococcus victoriae JSS0508, Metschnikowia pulcherrima JSS0502, Papiliotrema aurea JSS0505, Debaryomyces vanrijia JSS0509, Occultifur externus JSS0510, Filobasidium floriforme JSS0511 and Yamadazyma scolyti JSS0512 represented newly recorded yeast strains in Korea, and their microbiological characteristics were investigated. All of these unrecorded yeasts showed oval shape and also formed ascospores and pseudomycelia, except for Kwoniella mangroviensis JSS0500, Candida bombi JSS0503, and Metschnikowia pulcherrima JSS0502. Seven strains including Candida corydalis JSS0501 grew in vitamin-free medium, and 4 of the wild yeasts including Cryptococcus victoriae JSS0508 were halotolerant, i.e., capable of growing in 10% NaCl-containing yeast extract-peptone-dextrose (YPD) broth. Debaryomyces vanrijia JSS0509 was found to be a thermophilic yeast that grew at $37^{\circ}C$.

Fermentation Characteristics of Wild Grape (Vitis amurensis) Wine Prepared with Reed (Phragmites communis) Root (갈대뿌리를 첨가하여 제조한 산머루주의 발효 특성)

  • In, Man-Jin;Kim, Dong-Chung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.4
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    • pp.1528-1533
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    • 2010
  • This study investigated the fermentation characteristics of wild grape(Vitis amurensis) wine prepared with reed(Phragmites communis) root. The reed root was added to the fermentation of wild grape wine in the quantities of 0.5%, 1.0% and 2.0%. An addition of reed root during yeast fermentation of wild grape wine decreased the acidity and increased pH of fermentation culture, thus enhancing the consumption of sugar and production of ethanol. Especially, the addition of 2% reed root led to production of about 13% ethanol in 4 day of fermentation time. The sensory evaluation showed that the wild grape wine fermented with 2.0% reed root was most acceptable.

Isolation of Wild Killer Yeast from Traditional Meju and Production of Killer Toxin (재래식 메주로부터 야생 Killer 효모의 분리 및 Killer Toxin의 생산)

  • Lee, Jong-Su;Lee, Seong-Hun;Kim, Jae-Ho;Yu, Jin-Yeong
    • KSBB Journal
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    • v.14 no.4
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    • pp.434-439
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    • 1999
  • A wild yeast S-13 which has excellent killer toxin activity to gas-producing yeast of traditional Doenjang and Kochujang was selected among forty seven strains of Meju yeasts and identified as Hansenular capsulata S-13 by investigation of the morphological, cultural and physiological properties. The optimal conditions for the production of killer toxin were investigated. H. capsulata s-13 showed the higest killer toxin activities when it was cultured up to the late-log phase of 36 hr in YEPD medium (pH4.5) at $25^{\circ}C$ H. capsultara S-13 showed killer toxin activities to seven strains of industrial yeasts such as S. cerevisiae, C. veratilis and P. membranaefacieus.

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Construction of hsf1 Knockout-mutant of a Thermotolerant Yeast Strain Saccharomyces cerevisiae KNU5377 (고온내성 연료용 알코올 효모균주 Saccharomyces cerevisiae KNU5377에서 HSF1 유전자의 변이주 구축)

  • Kim Il-Sup;Yun Hae-Sun;Choi Hye-Jin;Sohn Ho-Yong;Yu Choon-Bal;Kim Jong-Guk;Jin Ing-Nyol
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.454-458
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    • 2006
  • HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

Identification of Wild Yeast Strains and Analysis of Their ${\beta}$-Glucan and Glutathione Levels for Use in Makgeolli Brewing

  • Kang, Sun Hee;Kim, Hye Ryun;Kim, Jae Ho;Ahn, Byung Hak;Kim, Tae Wan;Lee, Jang-Eun
    • Mycobiology
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    • v.42 no.4
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    • pp.361-367
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    • 2014
  • Makgeolli, also known as Takju, is a non-filtered traditional Korean alcoholic beverage that contains various floating matter, including yeast cells, which contributes to its high physiological functionality. In the present study, we assessed the levels of ${\beta}$-glucan and glutathione in various yeast strains isolated from traditional Korean Nuruk and selected a ${\beta}$-glucan- and glutathione-rich yeast strain to add value to Makgeolli by enhancing its physiological functionality through increased levels of these compounds. Yeast ${\beta}$-glucan levels ranged from 6.26% to 32.69% (dry basis) and were strongly species-dependent. Dried Saccharomyces cerevisiae isolated from Nuruk contained $25.53{\mu}g/mg$ glutathione, $0.70{\mu}g/mg$ oxidized glutathione, and $11.69{\mu}g/g$ and $47.85{\mu}g/g$ spermidine and L-ornithine monohydrochloride, respectively. To produce functional Makgeolli, a ${\beta}$-glucan- and glutathione-rich yeast strain was selected in a screening analysis. Makgeolli fermented with the selected yeast strain contained higher ${\beta}$-glucan and glutathione levels than commercial Makgeolli. Using the selected yeast strain to produce Makgeolli with high ${\beta}$-glucan and glutathione content may enable the production of functional Makgeolli.

Substitution of Asp-223 Residue to Leu in Yeast Alcohol Dehydrogenase and Coenzyme Specificity (효모 알코올 탈수소효소 아스파르트산-223 잔기의 루신으로 치환과 보조효소의 특이성)

  • Lee, Kang-Man;Ryu, Ji-Won
    • YAKHAK HOEJI
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    • v.36 no.5
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    • pp.469-473
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    • 1992
  • Yeast alcohol dehydrogenase (YADH) has an acidic residue that interacts with the 2'- and 3'-hydroxyl groups of the adenosine ribose of the $NAD^+$ coenzyme. The acidic residue of Asp-223 (according to horse liver alcohol dehydrogenase amino acid sequence) is supposed to determine the coenzyme specificity for $NAD^+$ rather than $NADP^+$. We mutated Asp-223 to leucine and the mutant YADH was expressed in yeast and characterized for the coenzyme specificity. The turnover numbers of mutant enzyme for $NAD^+$ and ethanol were decreased 3.5- and 4.8-fold compared to wild-type enzyme, respectively. Contrastively, catalytic specificity for $NADP^+$ was increased 13-fold. As a result, the mutant YADH also employed $NADP^+$ as a coenzyme.

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Isolation and characterization of unrecorded yeast species from Korea in the families Debaryomycetaceae and Piskurozymaceae

  • Lee, Sang Eun;Oh, Hye Jin;Kim, Myung Kyum
    • Journal of Species Research
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    • v.10 no.4
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    • pp.344-349
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    • 2021
  • The purpose of this study was to isolate and identify wild yeasts from soil of Gyeongju city, and Haemadipsa rjukjuana of Gageodo Island, characterizing unrecorded yeast strains from Korea. The molecular analysis of the D1/D2 domain of 26S rDNA of yeast was performed using the Basic Local Alignment Search Tool (BLAST). No official report exists describing these three species: one species in the genus Candida, one species in the genus Debaryomyces, and one species in the genus Solicoccozyma. Candida saitoana YL9, Debaryomyces fabryi YL1, and Solicoccozyma terrea 20g9-1 are recorded for the first time from Korea. All three strains were oval shaped and polar binding, while positive for glucose, ᴅ-xylose, and ᴅ-cellobiose. Morphological, physiological, and biochemical properties are described in the species descriptions.

Strain Improvement of Candida tropicalis for the Production of Xylitol: Biochemical and Physiological Characterization of Wild-type and Mutant Strain CT-OMV5

  • Rao Ravella Sreenivas;Jyothi Cherukuri Pavanna;Prakasham Reddy Shetty;Rao Chaganti Subba;Sarma Ponnupalli Nageshwara;Rao Linga Venkateswar
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.113-120
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    • 2006
  • Candida tropicalis was treated with ultraviolet (UV) rays, and the mutants obtained were screened for xylitol production. One of the mutants, the UV1 produced 0.81g of xylitol per gram of xylose. This was further mutated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and the mutants obtained were screened for xylitol production. One of the mutants (CT-OMV5) produced 0.85g/g of xylitol from xylose. Xylitol production improved to 0.87 g/g of xylose with this strain when the production medium was supplemented with urea. The CT-OMV5 mutant strain differs by 12 tests when compared to the wild-type Candida tropicalis strain. The XR activity was higher in mutant CT-OMV5. The distinct difference between the mutant and wild-type strain is the presence of numerous chlamvdospores in the mutant. In this investigation, we have demonstrated that mutagenesis was successful in generating a superior xylitol-producing strain, CT-OMV5, and uncovered distinctive biochemical and physiological characteristics of the wild-type and mutant strain, CT-OMV5.

Metabolite Profiling during Fermentation of Makgeolli by the Wild Yeast Strain Saccharomyces cerevisiae Y98-5

  • Kim, Hye Ryun;Kim, Jae-Ho;Ahn, Byung Hak;Bai, Dong-Hoon
    • Mycobiology
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    • v.42 no.4
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    • pp.353-360
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    • 2014
  • Makgeolli is a traditional Korean alcoholic beverage. The flavor of makgeolli is primarily determined by metabolic products such as free sugars, amino acids, organic acids, and aromatic compounds, which are produced during the fermentation of raw materials by molds and yeasts present in nuruk, a Korean fermentation starter. In this study, makgeolli was brewed using the wild yeast strain Saccharomyces cerevisiae Y98-5, and temporal changes in the metabolites during fermentation were analyzed by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. The resultant data were analyzed by partial least squares-discriminant analysis (PLS-DA). Various metabolites, including amino acids, organic acids, sugar alcohols, small peptides, and nucleosides, were obviously altered by increasing the fermentation period. Changes in these metabolites allowed us to distinguish among makgeolli samples with different fermentation periods (1, 2, 3, 6, 7, and 8 days) on a PLS-DA score plot. In the makgeolli brewed in this study, the amounts of tyrosine ($463.13{\mu}g/mL$) and leucine ($362.77{\mu}g/mL$) were high. Therefore, our results indicate that monitoring the changes in metabolites during makgeolli fermentation might be important for brewing makgeolli with good nutritional quality.