• Title/Summary/Keyword: yeast strain

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Molecular Cloning, Chromosomal Integration and Expression of the Homoserine Kinase gene THR1 of Saccharomyces cerevisiae (트레오닌 생합성에 관여하는 효모유전자 THR1의 클로님, 염색체통합 및 발현)

  • 최명숙;이호주
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.16-24
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    • 1991
  • The yeast gene THR1 encodes the homoserine kinase (EC 2.7.1.39: HKase) which catalyses the first step of the threonine specific arm at the end of the common pathway for methionine and threonine biosynthesis. A recombinant plasmid pMC3 (12.6 kilobase pairs, vector YCp50) has been cloned into E. coli HB101 from a yeast genomic library through its complementing activity of a thr1 mutation in a yeast recipient strain M39-1D. When subcloned into pMC32 (8.6kbp, vector YRp7) and pMC35 (8.3 kbp, vector YIp5), the HindIII fragment (2.7 kbp) of pMC3 insery was positive in the thrI complementing activity in both yeast and E. coli auxotrophic strains. The linearized pMC35 was introduced into the original recipient yeast strain and the mitotically stable chromosomal integrant was identified among the transformants. Through the tetrad analysis, the integration site of the pMC35 was localized to the region of THR1 structural gene at an expected genetic distance of approximately 11.1 cM from the ARG4 locus on the right arm of the yeast chromosome VIII. When episomically introduced into the auxotrophic cells and cultured in Thr omission liquid medium, the cloned gene overexpressed the HKase in the order of thirteen to fifteenfold, as compared with a wildtype. HKase levels are repressed by addition of threonine at the amount of 300 mg/l and 1, 190 mg/l for pMC32 and pMC3, respectively. Data from genetic analysis and HKase response thus support that the cloned HindIII yeast DNA fragment contains the yeast thr1 structural gene, along with necessary regulatory components for control of its proper expression.

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The Fermentation Characteristics of Saccharomyces cerevisiae F38-1 a Thermotolerant Yeast Isolated for Fuel Alcohol Production at Elevated Temperature (연료용 알콜의 고온발효를 위해 분리한 고온성 효모균주 Saccharomyces cerevisiae F38-1의 발효 특성)

  • 김재완;김상헌;진익렬
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.624-631
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    • 1995
  • The fermentation characteristics of Saccharomyces cerevisiae F38-1, a newly isolated thermotolerant yeast strain from a high temperature environment have been studied using a fermentation medium containing 20% glucose, 0.2% yeast extract, 0.2% polypeptone, 0.3% (NH$_{4}$)$_{2}$SO$_{4}$, 0.1% KH$_{2}$PO$_{4}$, and 0.2% MgSO$_{4}$ without shaking at 30$\circ$C to 43$\circ$C for 5 days. The fermentability was over 90% at 30$\circ$C, 88% at 37$\circ$C, 77% at 40$\circ$C and 30% at 43$\circ$C. A similar fermentation result was obtained at pH between 4 and 6 at 30$\circ$C and 40$\circ$C. Aeration stimulated the growth of the strain at the beginning of the fermentation, but it reduced alcohol production at the end of alcohol fermentation. Optimal glucose concentration was determined to be between 18 and 22% at 40$\circ$C as well as 30$\circ$C, but the growth was inhibited at the glucose concentration of over 30%. A fermentability of over 90% was observed at 40$\circ$C in 2 days when the medium was supplemented by 2% yeast extract. A higher inoculum size increased the initial fermentation rate, but not the fermentation. A fermentability of over 90% was achieved in 2 days at 40$\circ$C in a fermentor experiment using an optimized medium containing 20% glucose and 1% yeast extract.

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Toluene Removal and Microbial Growth of Candida tropicalis Immobilized with Polymer Media in Airlift Bioreactors (효모 Candida tropicalis 고정화 담체를 이용한 Airlift 미생물반응기의 톨루엔 제거 및 미생물 성장)

  • Namgung, Hyeong-Kyu;Song, JiHyeon;Jung, Mi-Young;Hwang, Sun-Jin
    • 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.

Biosynthetic Pathway of Indole-3-Acetic Acid in Basidiomycetous Yeast Rhodosporidiobolus fluvialis

  • Bunsangiam, Sakaoduoen;Sakpuntoon, Varunya;Srisuk, Nantana;Ohashi, Takao;Fujiyama, Kazuhito;Limtong, Savitree
    • Mycobiology
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    • v.47 no.3
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    • pp.292-300
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    • 2019
  • IAA biosynthetic pathways in a basidiomycetous yeast, Rhodosporidiobolus fluvialis DMKU-CP293, were investigated. The yeast strain showed tryptophan (Trp)-dependent IAA biosynthesis when grown in tryptophan supplemented mineral salt medium. Gas chromatography-mass spectrometry was used to further identify the pathway intermediates of Trpdependent IAA biosynthesis. The results indicated that the main intermediates produced by R. fluvialis DMKU-CP293 were tryptamine (TAM), indole-3-acetic acid (IAA), and tryptophol (TOL), whereas indole-3-pyruvic acid (IPA) was not found. However, supplementation of IPA to the culture medium resulted in IAA peak detection by high-performance liquid chromatography analysis of the culture supernatant. Key enzymes of three IAA biosynthetic routes, i.e., IPA, IAM and TAM were investigated to clarify the IAA biosynthetic pathways of R. fluvialis DMKU-CP293. Results indicated that the activities of tryptophan aminotransferase, tryptophan 2-monooxygenase, and tryptophan decarboxylase were observed in cell crude extract. Overall results suggested that IAA biosynthetic in this yeast strain mainly occurred via the IPA route. Nevertheless, IAM and TAM pathway might be involved in R. fluvialis DMKU-CP293.

Isolation and Identification of Wild Yeast and Its Use for the Production of Grapewine (야생 효모의 분리.동정 및 이를 이용한 포도주 제조)

  • Kim, Jung-In;Lee, Nam-Keun;Hahm, Young-Tae
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.217-221
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    • 2007
  • The domestic cultured Campbell's Early and Geubong grapes were fermented far the production of red wines with the isolated wild yeast Saccharomyces cerevisiae IJ850. For the isolation of wild yeast, Geubong and Campbell's Early grapejuices were naturally fermented at room temperature for 6 days without adding stater culture. The strain isolated from Geubong which has 1.8 times higher fermentative ability than the strains isolated Campbell Early was selected. The selected strain was identified by using 26S rDNA sequencing. The strain showed 99.7% of similarity with Saccharomyces cerevisiae and thus identified as Saccharomyces cerevisiae IJ850. It was investigated the fermentative ability as the start culture. For the production of grapewine, the final sugar concentrations of grapejuices were adjusted to the $25^{\circ}Brix$ with anhydrous glucose. The grapejuices were fermented at room temperature for 10 days in the air-locked bottles filled with $CO_2$ gas. The final yield and alcohol concentration of Campbell's Early and Geubong grapewines fermented with the isolated wild yeast were 80.8%, 11.0% and 87.8%, 13.0%, respectively. Between the isolated wild yeast S. cerevisiae IJ850 and the commercial yeast S. cerevisiae EC1118, total acidities of grapewines produced with wild yeast were lower than those produced with the commercial yeast. The pH values and the values of color analysis of grapewines produced with both strains were similar. The total phenol contents of campbell's Early and Geubong wines produced with the isolated yeast and the commercial yeast were obtained in the range of 75 to 125mg/L. In conclusion, S. cerevesiae IJ850 isolated from the domestic cultured Geubong grape is able to use to produce grapewines as stater culture.

Studies on the Petroleum hydrocarbon-utilizing Micro-organisms(Part 2) - On the Production of Single Cell Protein from Petroleum hydrocarbon with a yeast strain - (석유 탄화수소 이용 미생물에 관한 연구 (제 2 보) - 효모를 이용한 석유탄화수소로 부터 단백질 생산에 관하여 -)

  • Lee, Ke-Ho;Shin, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.9-18
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    • 1971
  • In order to obtain basic information on the production of single cell protein from petroleum, more than 400 yeast strains were isolated from various soil samples in Korea utilizing petroleum hydrocarbon as the sole carbon source. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimal culture condition was searched in the flasks shaken throughout the procedure. And the growing characteristics for the selected yeast strain and chemical analysis of the yeast cell component were carried out. The results obtained were as follows: 1. The selected yeast strain was identified as Candida curvata and we named it Candida curvata-SNU 70. 2. The composition of the medium proposed for the present yeast strain is: Light Gas Oil 30ml, Urea 400mg, Ammonium sulfate 100mg, Potasium phosphate (monobasic) 670mg, Sodium phosphate (dibasic) 330mg, Magnesium sulfate 500mg, Calcium carbonate 3g, Yeast extract 50mg, Tween 20 0.05ml, Tap water 1,000ml. 3. Other culture conditions employed for the yeast were pH 5.5-7.0, temp. $30^{\circ}C$ under an affluent aerobic state. 4. Addition of light gas oil in portions to the culture media as the growth proceeded was more effective, especially in the cultivation on the higher oil concentration media. 5. Studies on the propagation of the yeast cells in the light gas oil medium revealed that the yeast has the lag phase lasted 16 hours and the logarithmic growth phase covered 16 to 28 hours. The specific growth rate was about $0.22\;hr^{-1}$ and doubling time was 3.2 hrs. during the logarithmic growth phase. 6. Under the cultural condition employed, the cell yield against the amount of light gas oil (wt%) was 16.1% and the protein content of the dried yeast cells was 48.4%.

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Study of Cytotoxicity of an Actinomycete Isolated in Korea (토양에서 분리한 방선균의 세포 독성에 관한 연구)

  • Park, Joon-Koo;Choi, Boung-Don;Kim, Seung-Chul;Ryeom, Kon
    • Environmental Analysis Health and Toxicology
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    • v.8 no.3_4
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    • pp.7-12
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    • 1993
  • An Actinomycete strain isolated from Mt. Dea-Dun had a strong antifungal activity. The culture brith produced by isolated strain showed only antifungal activity against fungi with the exception of yeast and bacteria. It was heat stable, dissolved in ehtylacetate. The concentrated antifungal agent showed cytotoxicity against HEP-2 and HeLa as tumor cell line, and showed weak cytotoxicity against VERO 36 as normal cell line. Morphological and physiological characteristics were tested with isolated strain. The spore color of isolated strain was gray. It had a short chain and produced brown colored lytic substance in yeast extract-malt agar. The cell wall of isolated strain was composed of meso-DAP, and we suggested it as genus Actinomadura. In the existing of chemical inhibitor, isolated strain grew on the condition of 0.0001% crystal violet, 0.1% phenol, 0.01% sodium azide and 10% sodium chloride. Carbon utilization of isolated strain was shown that glucose, sucrose, manitol and sodium citrate were well utilized.

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Isolation of Novel Alkalophilic Bacillus alcalophilus subsp. YB380 and the Characteristics of Its Yeast Cell Wall Hydrolase

  • Yeo, Ik-Hyun;Han, Suk-Kyun;Yu, Ju-Hyun;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.501-508
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    • 1998
  • An alkalophilic mi.croorganism (strain YB380), which produces yeast cell wall hydrolase extracellulary, was isolated from Korean soil. The rod-shaped cells were 0.3~0.4 by 2~4${\mu}{\textrm}{m}$ long, motile, aerobic, gram-positive, and spore-forming. The color of the colony was light yellow. The temperature range for growth at pH 9.0 was 25 to $45{\circ}C, with optimum growth at $35{\circ}C. The pH range for growth at $35{\circ}C was 8 to 11 with an optimum pH of 9.0. Therefore, the strain YB380 is an obligate alkalophile. The 16S rRNA of strain YB380 has a 99% sequence similarity with that of Bacillus alcalophilus. On the basis of physiological properties, cell wall fatty acid composition, and phylogenetic analysis, we propose that the isolated strain is Bacillus alcalophilus. The yeast cell wall hydrolase from Bacillus alcalophilus subsp. YB380 has been purified and partially characterized. The molecular weight was estimated to be 27,000 daltons with an optimum temperature and pH of $60{\circ}C and 9.0, respectively. The N-terminal amino acid sequence of the enzyme was analyzed as Gln- Thr- Val- Pro- Trp- Gly- Ile- Asn- Arg- Val.

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Construction of an Industrial Brewing Yeast Strain to Manufacture Beer with Low Caloric Content and Improved Flavor

  • Wang, Jin-Jing;Wang, Zhao-Yue;Liu, Xi-Feng;Guo, Xue-Na;He, Xiu-Ping;Wense, Pierre Christian;Zhang, Bo-Run
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.767-774
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    • 2010
  • In this study, the problems of high caloric content, increased maturation time, and off-flavors in commercial beer manufacture arising from residual sugar, diacetyl, and acetaldehyde levels were addressed. A recombinant industrial brewing yeast strain (TQ1) was generated from T1 [Lipomyces starkeyi dextranase gene (LSD1) introduced, ${\alpha}$-acetohydroxyacid synthase gene (ILV2) disrupted] by introducing Saccharomyces cerevisiae glucoamylase (SGA1) and a strong promoter (PGK1), while disrupting the gene coding alcohol dehydrogenase (ADH2). The highest glucoamylase activity for TQ1 was 93.26 U/ml compared with host strain T1 (12.36 U/ml) and wild-type industrial yeast strain YSF5 (10.39 U/ml), respectively. European Brewery Convention (EBC) tube fermentation tests comparing the fermentation broths of TQ1 with T1 and YSF5 showed that the real extracts were reduced by 15.79% and 22.47%; the main residual maltotriose concentrations were reduced by 13.75% and 18.82%; the caloric contents were reduced by 27.18 and 35.39 calories per 12 oz. Owing to the disruption of the ADH2 gene in TQ1, the off-flavor acetaldehyde concentrations in the fermentation broth were 9.43% and 13.28%, respectively, lower than that of T1 and YSF5. No heterologous DNA sequences or drug resistance genes were introduced into TQ1. Hence, the gene manipulations in this work properly solved the addressed problems in commercial beer manufacture.

Strategy for Prevention of Weakly Flocculating Characters in Bottom Brewing Yeast Strains

  • Cheong, Chul;Wackerbauer, Karl;Kang, Soon-Ah
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.558-563
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    • 2008
  • To prevent weakly flocculating characters of bottom brewing yeast during first fermentation, various technical investigations were carried out using strain of Saccharomyces cerevisiae. It appeared that the propagation at $10^{\circ}C$ promoted the molecular structure and biochemical composition of cell wall in favor of flocculation. The yeast grown at $20^{\circ}C$ by addition of zinc ion also had a stimulating effect on flocculation behavior during first fermentation cycle. The zinc ion did not influence directly on the changes of cell wall in favor of stronger flocculence. The increased fermentation activity of yeast due to addition zinc ion was rather responsible for the intensified flocculation capacity. It was concluded that the weakly flocculating characters of bottom brewing yeast during first fermentation can be solved by using yeast propagated at $10^{\circ}C$ or by means of yeast by addition of zinc ion during propagation.