• 제목/요약/키워드: non-Saccharomyces

검색결과 127건 처리시간 0.027초

Molecular Cloning and Analysis of Sporulation-Specific Glucoamylase (SGA) Gene of Saccharomyces diastaticus

  • Kang, Dae-Ook;Hwang, In-Kyu;Oh, Won-Keun;Lee, Hyun-Sun;Ahn, Soon-Cheol;Kim, Bo-Yeon;Mheen, Tae-Ick;Ahn, Jong-Seog
    • Journal of Microbiology
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    • 제37권1호
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    • pp.35-40
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    • 1999
  • Sporulation-specific glucoamylase (SGA) gene was isolated from genomic library of Saccharomyces diastaticus 5114-9A by using glucoamylase non-producing mutant of S. diastaticus as a recipient. When the glucoamylase activities of culture supernatant, periplasmic, and intracellular fraction of cells transformed with hybrid plasmid containing SGA gene were measured, the highest activity was detected in culture supernatant. SGA produced by transformant and extracellular glucoamylase produced by S. diastaticus 5114-9A differed in enzyme characteristics such as optimum temperature, thermostability, and resistance to SDS and urea. But the characteristics of SGA produced by sporulating yeast cells and vegetatively growing transformants were identical.

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Incapability of Utilizing Galactose by pgs1 Mutation Occurred on the Galactose Incorporation Step in Saccharomyces cerevisiae

  • Rho, Min-Suk;Su, Xuefeng;Lee, Yoon-Shik;Kim, Woo-Ho;Dowhan, William
    • Journal of Microbiology and Biotechnology
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    • 제16권1호
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    • pp.84-91
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    • 2006
  • A Saccharomyces cerevisiae pgs1 nulI mutant, which is deficient with phosphatidyl glycerol (PG) and cardiolipin (CL) biosynthesis, grows well on most fermentable carbon sources, but fails to grow on non-fermentable carbon sources such as glycerol, ethanol, and lactate. This mutant also cannot grow on galactose medium as the sole carbon source. We found that the incorporation of $[^{14}C]-galactose$, which is the first step of the galactose metabolic pathway (Leloir pathway), into the pgs 1 null mutant cell was extremely repressed. Exogenously expressed PGS1 (YCpPGS1) under indigenous promoter could completely restore the pgs1 growth defect on non-fermentable carbon sources, and dramatically recovered $[^{14}C]-galactose$ incorporation into the pgs1 mutant cell. However, PGS1 expression under the GALl promoter $(YEpP_{GAL1}-PGS1myc)$ could not complement pgs1 mutation, and the GAL2-lacZ fusion gene $(YEpP_{GAL2}-lacZ)$ also did not exhibit its $\beta-galactosidase$ activity in the pgs1 mutant. In wild-type yeast, antimycin $A(1\;{\mu}g/ml)$, which inhibits mitochondrial complex III, severely repressed not only the expression of the GAL2-lacZ fusion gene, but also uptake of $[^{14}C]-galactose$. However, exogenously expressed PGS1 partially relieved these inhibitory effects of antimycin A in both the pgs1 mutant and wild-type yeast, although it could not basically restore the growth defect on galactose by antimycin A. These results suggest that the PGSI gene product has an important role in utilization of galactose by Gal genes, and that intact mitochondrial function with PGS1 should be required for galactose incorporation into the Leloir pathway. The PGS1 gene might provide a clue to resolve the historic issue about the incapability of galactose with deteriorated mitochondrial function.

The Viability Change of Yeast Cell Responding to Trehalose Accumulation and Maintaining Neutral Trehalase Activity under Extracellular pH Acidified by $H_2SO_4$

  • Jin, Ingnyol;Yun, Haesun;Paik, Sanhkyoo;Kim, Ilsup;Sohn, Ho-Yong
    • Journal of Life Science
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    • 제12권2호
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    • pp.47-52
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    • 2002
  • Saccharomyces cerevisiae KNU5377 (KNU5377) and S. cerevisiae ATCC24858 (ATCC24858) were exposed to $H_2SO_4$ as a stress, which was added at various concentrations to a YPD media. The growth of KNU5377 was reduced to approximately 60% in the YPD media containing 40 nm sulfuric acid when compared to the non-stressed condition. When their growth was monitored during an overnight culture, two strains, KNU5377 and ATCC24858, could not grow when exposed to over 50 mM of sulfuric acid. After a short exposure to this acid for 1 h, KNU5377 exhibited stronger resistance against $H_2SO_4$ than ATCC24858. The neutral trehalase activity of KNU5377 unchanged despite under various concentrations of $H_2SO_4$. In contrast, It at of ATCC24858 was much low at higher $H_2SO_4$concentrations. Trehalose, a non-reducing disaccharide, was maximally accumulated after a short exposure to 60 nm $H_2SO_4$ for KNU5377, but it was reduced under more severe stressful conditions. These results suggest that KNU5377 should modulate the trehalose concentrations under the severe stress condition of high sulfuric acid concentrations. The most highly induced protein in the KNU5377 exposed to sulfuric acid was found to be an approximately 23 kDa protein, which was revealed to be the 605 large subunit ribosomal protein, Ll3 by FASTA search results.

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골담초 효모 발효 추출물의 피부 미백 효능에 관한 연구 (Skin Whitening Effects of Caragana sinica Rehder Extract Fermented by Saccharomyces cerevisiae KCTC 7913)

  • 석지현;이선영;채은정;최신욱
    • 대한화장품학회지
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    • 제36권3호
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    • pp.207-213
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    • 2010
  • 본 연구는 미생물에 의한 골담초(Caragana sinica Rehder) 발효 추출물의 항산화 효과 및 피부 미백 효능에 관한 연구로 골담초에 S. cerevisiae KCTC 7913를 첨가하여 발효를 통해 얻어진 발효 추출물을 B16F10 멜라닌 세포에 처리한 결과, 농도의존적으로 멜라닌 생성 억제 효능이 증가한 것으로 확인하였으며, 추출물 내 유효 성분인 resveratrol의 함량이 발효 전의 추출물에 비해 증가함을 확인하였다. 멜라닌 생성 과정의 주요 단백질인 tyrosinase의 활성 억제 또한 골담초 추출물에 비해 골담초 발효 추출물의 효능이 우수함을 확인하였으며, 골담초 발효 추출물의 피부 자극 또한 일어나지 않는 것으로 확인되어 피부에 안전한 화장품 원료로서의 사용이 가능할 것으로 사료된다.

Analysis of Biological Effect of DBD-type Non-thermal Atmospheric Pressure Plasma on Saccharomyces Cerevisiae

  • Park, Gyung-Soon;Baik, Ku-Yeon;Kim, Jung-Gil;Kim, Yun-Jung;Lee, Kyung-Ae;Choi, Eun-Ha;Uhm, Hwan-Sup;Jung, Ran-Ju;Cho, Kwang-Sup
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2011년도 제41회 하계 정기 학술대회 초록집
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    • pp.337-337
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    • 2011
  • Application of plasma technology on microbial sterilization has been frequently studied. In spite of accumulating number of studies, many have been focused on bacteria. Reports on eukaryotic yeasts and filamentous fungi are limited. In addition, mechanism of plasma effect still needs to be clarified. In this study, we analyzed the effect of non-thermal atmospheric pressure plasma on the budding yeast, Saccharomyces cerevisiae using DBD-type device. When yeast cells were exposed to plasma (at 2 mm distance) and then cultured on YPD-agar plate, number of cells survived (shown as colony) were reduced proportionally to exposure time. More than 50% reduction in number of colonies were observed after twice exposure of 5min. each. Colonies much smaller than those of control (no plasma exposure) were appeared after twice exposure of 5 min. each. It seems that small colonies are resulted from delayed cell growth due to the damage caused by plasma treatment. Microscopic analysis demonstrates that yeast cells treated with plasma for 5 min. twice have more rough and shrinked shape compared to oval shape with smooth surface of control.

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복분자 와인 제조를 위한 바이오제닉 아민 비생성 효모의 선별 및 통계학적 기법을 이용한 배양조건 최적화 (Screening of Biogenic Amine Non-Producing Yeast and Optimization of Culture Conditions Using Statistical Method for Manufacturing Black Raspberry Wine)

  • 양희종;정수지;정성엽;허주희;정도연
    • 한국식품영양과학회지
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    • 제44권4호
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    • pp.592-601
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    • 2015
  • 복분자 와인의 제조를 위하여 복분자 과실 및 엑기스로부터 야생효모 Saccharomyces cerevisiae BA29를 분리 및 동정하였으며, 생화학적 특성 및 biogenic amine 생성 여부, 배양학적 특성 및 알코올 발효능과 알코올, 당, 아황산 저항성을 조사하였다. 또한 S. cerevisiae BA29의 산업적 공정 적용을 위한 균체량 증가를 위하여 통계학적 방법인 반응표면분석법을 사용하여 배양 배지 조성의 최적화를 수행하였다. 실험계획법은 중심합성계획을 사용하여 모델을 설정하였고, 산업적 공정 적용 시 비용 대비 효율성이 높은 molasses를 대체 탄소원으로 사용하여 실험을 수행하였다. 통계프로그램을 이용하여 분석한 결과 최대 균체 성장을 위한 배지 조성으로는 molasses 200 g/L, peptone 30 g/L, yeast extract 40 g/L로 예측되었으며, 이때의 최대 균체량은 20.6565 g/L로 예측되었다. 모델의 검증실험 결과 기본 배양 배지와 비교하였을 때 6.39 g/L에서 $20.9167{\pm}0.7925g/L$로 약 3.27배 증가하였다. 최종적으로 S. cerevisiae BA29를 사용하여 복분자 와인을 제조한 결과 20.33%의 알코올 생성능을 나타냈다. 이로써 복분자 과실로부터 분리한 효모 S. cerevisiae BA29를 이용한 우수한 복분자 와인 제조의 가능성을 확인하였다.

Rare Mating에 의한 양초효모에서의 glucoamylase 발현 균주 HCS 선별 및 특성 (Characterization of Brewing Yeast Expressing Glucoamylase Selected by Rare Mating.)

  • 최병주;장금일;김광엽
    • 한국미생물·생명공학회지
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    • 제29권4호
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    • pp.212-220
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    • 2001
  • S. cerevisiae HBC52와 S, diataticus K114 의 rare mating 에 의해 개발된 HCS 균주들은 크기가 약 $13\mu\textrm{m}$ karyotype 분석결과 K114 균주에만 있는 약 1150kb 분자량을 가지는 염색체 band를 유지하였으며 전분을 분해하여 halo 를 형성하였다. Glucoamylase 활성은 약 2.7~3.4 unti/ml 를 가진 균주임이 밝혀졌으며 당 발효실험과 응집성 실험을 수행한 결과 HBC52 균주와 유사한 당 발효특성을 보이고 응집성 특성도 약응집성의 floculation type으로 비슷하였다. 그리고 HCS 균주의 포자형성과 피막형성 유무 실험에서는 양조효모인 HBC52 균주와 같이 포자가 형성되지 않았으며, 피막도 형성되지않았다. 균주들의 최종당도 실험은 HBC52균주가 약 68%의 발효수준을 나타냈고, HCS 균주들은 이 보다 높은 76~78%의 수준을 보였따. 즉 HBC52 균주가 최종당도($ 2.00^{\circ}$P)를 보인 반면 HCS 균주들은 ($0.7~0.93^{\circ}$P)를 보이는 결괄르 나타내어 맥주양조에서 low carbohydrate beer를 생산할 수있음이 확인되었다.

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연속식 고정화 및 현탁 세포 생물 반응기에 의한 에탄을 생성중 지질 첨가 영향 (The Roles of Lipid Supplements in Ethanol Production Using a Continuous Immobilized and Suspended Cell Bioreactor)

  • 길광훈
    • Applied Biological Chemistry
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    • 제39권1호
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    • pp.1-8
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    • 1996
  • Saccharomyces cerevisiae의 고정화 및 현탁 세포로 구성된 연속식 생물반응기에서 에탄올 생성시 지질 첨가 영향을 연구하였다. 여러가지 배양조건하에서 에탄을 생산량 및 현탁 세포의 알코올 탈수소효소(alcohol dehydrogenase, ADH)의 비활성도를 측정하였다. 무통기 조건하에서 ergosterol과 oleic acid를 세포 배양액에 첨가하였을때, 에탄을 생산량과 균체 생육이 현저히 증가하였으나, 알코올 탈수소효소의 비활성도는 영향을 받지 않았다. 특히 무통기 조건 및 통기 조건하에서 얻어진 현탁 세포간의 알코올 탈수소효소의 비활성도는 차이가 없었다. 계면활성제 첨가시에도 에탄올 생성, 균체 생육, 알코올 탈수소효소의 비활성도가 크게 증가하였다. 고농도($40\;g/{\ell}$ 이상) 에탄올에 노출된 세포배양액에 ergosterol과 oletic acid 첨가시에도 에탄올 생성량, 균체 생육, 알코올 탈수소효소의 비활성도가 증가하였으나, 계면활성제 첨가시에는 효과가 없었다. 따라서, 지질 첨가효과는 저농도 에탄을 조건에 비해 고농도 에탄을 존재시 크게 작용하였다. 여러가지 매양조건에서 얻어진 현탁 세포의 알코올 탈수소효소의 isozyme patteren을 전기영동법에 의해 조사한 결과 ADH I으로 추정되는 한개의 isozyme만이 확인되었으며, isozyme의 이동거리는 세포의 배양조건에 따라 약간의 차이가 있었다. 에탄올 생성량과 알코올 탈수소효소의 비활성도사이의 상관관계는 성립되지 않았으며, 알코올 탈수소효소의 비활성도보다는 균체량이 에탄올 생성에 더 중요한 인자로 작용하였다.

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재조합효모 배양에서 비이온성 계면활성제가 외래 Glucoamylase 생산 및 분비에 미치는 영향

  • 차형준;유영제
    • 한국미생물·생명공학회지
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    • 제24권6호
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    • pp.712-716
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    • 1996
  • The effects of non-ionic surfactants (Triton X-100 and Tween 80) on cloned glucoamylase production and secretion in recombinant Saccharomyces cerevisiae culture were studied. Even though the extracellular glucoamylase activity was increased by addition of Tween 80 due to the increase of the cell mass, Tween 80 did not play a role in the increase of glucoamylase secretion. On the addition of Triton X-100 addition, the secretion efficiency was increased while the cell growth was inhibited. Triton X-100 was added to the culture broth after 24 hr of culture to minimize the inhibition of the cell growth, and consequently the glucoamylase activity in the culture broth was increased by 12%.

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Calcium Alginate에 포괄된 Yeast Invertase의 고정화 효소에 관한 연구 (II. 고정화 효모의 효소학적 특성) (Calcium Alginate-entrapped Yeast Whole-cell Invertase (II. Enzymatic Properties of the Immobilized Cells))

  • 방병호;이상건;양철영
    • 한국식품영양학회지
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    • 제2권2호
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    • pp.14-20
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    • 1989
  • A strain of Saccharomyces cerevisiae BY-366 was isolated to produce a strong sucrose-hydrolyzing enzyme. After entrapment of yeast cell invertase with alginate, enzymatic properties of immobilized cells were investigated. The results are as follows. 1. The optimum pH of invertase in immobilized cells and non- immobilized cells was 6.0 and 5.0, and pH stability of invertase in immobilized cells and non- immobilized cells was 6.0 and 5.0, respectively. 2 Activation energy of immobilized cells was 4.7 kcal/mol. 3 The immobilized preparation exhibited high resistance to heat and urea Induced denaturation. 4, The bead size less than 2 mm in diameter was desirable. 5. In spite of repeated use, the enzyme activity of immobilized cells was inhibited slightly in batch reaction, and a small column of the immobilized preparation could hydrolyze relatively high concentration of sucrose almost quantitatively to more than 6 days.

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