• 제목/요약/키워드: yeast mating

검색결과 49건 처리시간 0.029초

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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산업용 효모 Hybrid의 선별을 위한 우성선별표지로서의 Aureobasidin A 내성유전자의 이용 (The Use of Aureobasidin A Resistant Gene as the Dominant Selectable Marker for the Selection of Industrial Yeast Hybrid)

  • 전한택;박은미;김근
    • 한국미생물·생명공학회지
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    • 제39권2호
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    • pp.111-118
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    • 2011
  • 교배와 원형질체 융합을 통한 배수체인 야생형 산업균주의 개발을 위하여, hybrid의 선별표지로서 우성의 선별표지인 aureobasidin A 내성이 사용될 수 있는 지를 알아보고자 하였다. 선별배지에서 aureobasidin A의 최적농도는 야생형 균주인 경우 SD와 YPD 배지에서는 0.5 ${\mu}g$/mL 이상이었고, SG와 YPG에서는 0.2-0.3 ${\mu}g$/mL 이었다. 한편 호흡결여돌연변이주는 야생형 균주보다 훨씬 높은 농도의 aureobasidin A에도 내성이 있음을 나타내었다. 우리는 K114/YIP균주의 전분분해 능력이 배수체 야생형 산업 균주에 전달 될 수 있는지를 이 방법을 통하여 관찰하였다. 반수체 영양요구성 균주 K114/YIP에 aureobasidin에 대한 내성을 부여하는 pAUR112가 도입된 균주와 야생형 균주 KL 혹은 C6와의 rare-mating 후 aureobasidin A 함유 배지에서 성장한 hybrid를 분리할 수 있었다. Hybrid는 전분분해 능력을 함유하고 있었을 뿐 아니라 두 양친의 특성을 동시에 지녔으며, 전자현미경 관찰 결과에서도 hybrid는 양친주의 특성을 모두 갖는 것으로 나타났다.

이담자 효모균의 성분화과정에서 막단백질 중 $\Ca^{2+}$-ATPase와 trigger peptidase(TPase)의 상호관계 (Relation of $\Ca^{2+}$-ATPase and trigger peptidase(TPase) that are Membrane Proteins in a Differentiation Process on Heterobasidiomycerous Yeast)

  • 정영기;이태호;정경태
    • 한국미생물·생명공학회지
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    • 제22권1호
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    • pp.1-6
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    • 1994
  • We have studied the relation between Ca$^{2+}$-ATPase and trigger peptidase(TPase) which are membeane protein well known as their significant role for signal transduction of mating pheromone in heterobasidiomycetous yeast. Rhodosporidium toruloides. We found out that there were Ca $^{2+}$-ATPase and TPase together in isolated calmodulim binding protein(CBP), usion calmodulin affinity column chromatography after solubilization of mation type a cell membrane protein, and that the dependence of enzyme activity of both the enzymes on Ca$^{2+}$, phospholipid and nonionic detergent are similar. However, Ca$^{2+}$-ATPase hed quite absolute dependence on calmodulin and, on the other hand, TPase didn't have any dependence. Judging from the fact that there are both enzymes in CBP which the dependence of calmodulin are quite different, we found out that both enzymes were made to their compound and existed in mating type a cell membrane.

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반수체 Saccharomyces diastaticus의 에탄올내성 증진 (Improvement of Ethanol-Tolerance of Haploid Saccharomyces diastaticus)

  • 송상호;김근;이민웅
    • 한국미생물·생명공학회지
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    • 제22권6호
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    • pp.584-592
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    • 1994
  • Several mutation procedures have been compared to obtain an ethanol-tolerant Saccha- romyces diastaticus strain secreting glucoamylase. These procedures include spontaneous mutation, EMS treatment, UV irradiation, and combination of EMS treatment and UV irradiation. All these methods were followed by adaptation of the yeast cells to gradually higher ethanol concentration. Among these procedures, the combined method of EMS treatment and UV irradiation gave the promising result, i.e. the ethanol tolerance of the yeast increased from 11.5%(v/v) to 14.0%(v/v). Respiratory deficient petite mutants of industrial and ethanol-tolerant yeast strains have been isola- ted and hybridized with haploid S. diastaticus strains. The resulting hybrids showed increased ethanol tolerance and starch-fermentability.

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Production of Cellulosic Ethanol in Saccharomyces cerevisiae Heterologous Expressing Clostridium thermocellum Endoglucanase and Saccharomycopsis fibuligera β-glucosidase Genes

  • Jeon, Eugene;Hyeon, Jeong-eun;Suh, Dong Jin;Suh, Young-Woong;Kim, Seoung Wook;Song, Kwang Ho;Han, Sung Ok
    • Molecules and Cells
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    • 제28권4호
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    • pp.369-373
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    • 2009
  • Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.

Positive Charge of Arginine Residues on Histone H4 Tail Is Required for Maintenance of Mating Type in Saccharomyces cerevisiae

  • Yeom, Soojin;Oh, Junsoo;Lee, Eun-Jin;Lee, Jung-Shin
    • Journal of Microbiology and Biotechnology
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    • 제28권9호
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    • pp.1573-1579
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    • 2018
  • Transcriptional gene silencing is regulated by the chromatin structure, which is by various factors including histones. Saccharomyces cerevisiae contains transcriptionally silenced regions such as telomeric regions and hidden mating (HM) loci. The positively-charged amino acids on the histone H4 tail were reported to be critical for the telomeric silencing in yeast, by interacting with Dot1, a specific methyltransferase for the $79^{th}$ lysine on histone H3. However, Dot1 did not affect gene silencing within HM loci, but whether the positively-charged amino acids on the H4 tail affect HM silencing has not been defined. To elucidate the function of the H4 tail on HM silencing, we created several MATa-type yeast strains bearing the substitution of arginine with alanine or lysine on the histone H4 tail and checked the sensitivity of MATa-type yeast to alpha pheromone. The arginine point mutants substituted by alanine (R17A, R19A, and R23A) did not show sensitivity to alpha pheromone, but only two arginine mutants substituted by lysine (R17K and R19K) restored the sensitivity to alpha pheromone-like wild type. These data suggested that the basic property of arginine at $17^{th}$ and $19^{th}$ positions in the histone H4 tail is critical for maintaining HM silencing, but that of the $23^{rd}$ arginine is not. Our data implicated that the positive charge of two arginine residues on the histone H4 tail is required for HM silencing in a manner independent of Dot1.

효모에서 활성형의 곤충유래 항균펩티드 defensin의 발현 (Expression of Biologically Active Insect-Derived Antibacterial Peptide, Defensin, in Yeast)

  • 강대욱;안순철;김민수;안종석
    • 생명과학회지
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    • 제12권4호
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    • pp.477-482
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    • 2002
  • 효모 glucoamylase의 promoter와 분비신호서열 그리고 MF$\alpha$1의 prosequence를 이용하여 곤충 defensin을 S. cerevisiae 2805에서 항균활성을 보유한 형태로 발현 및 분비하는데 성공하였다. 발현된 defensin의 대부분이 세포 외로 분비되어 거의 모든 항균활성이 배양 상등액에 존재하였다. 이것은 S. cerevisiae에서 발현된 defensin이 glucoamylase의 분비신호서열과 MF$\alpha$1의 prosequenre에 의해 효율적으로 processing되어 분비됨을 시사한다. Defensin의 다른 미생물에 대한 항균활성을 조사한 결과 병원균인 St. aureus와 L. monocytogenes에 대해서도 항균활성이 존재하였다.

Rhodosporidium toruloides의 접합형 A 세포내 Invertase의 정제 (Purification of Internal Invertase in Rhodosporidium toruloides Mating Type A Cells)

  • 정영기;류병호
    • 한국식품영양과학회지
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    • 제21권6호
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    • pp.725-730
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    • 1992
  • 이담자 효모균 Rhodosporidium toruloides의 mating type A 세포에서 세포내 invertase를 정제하였다. 세포내 invertase는 배양 세포의 파쇄액을 산침전 시킨 후 그 상등액으로부터 DEAE-Sephadex A-50, SP-Sephadex C-50 column chromatography와 Sephadex G-200 gel fitration 등의 과정을 거쳐 polyacrylamide gel disc 전기영동상 단일 효소 단백질까지 정제되었다. 정제효소의 분자량은 gel filtration에 의하여 90,000이었고, SDS-PAGE상에서는 22,000 dalttons에서 단일 band를 보여 단일종의 subunit가 4개로 구성된 단백질로 추정된다.

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Purification and Characterization of Internal Invertase in Rhodosporidum toruloides Mating Type a Cells

  • Jeong, Youn-Kee;Cho, Kyung-Soon;Lee, Tae-Ho;Ryu, Beung-Ho
    • Preventive Nutrition and Food Science
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    • 제2권3호
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    • pp.250-254
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    • 1997
  • The internal invertase of Rhodosporidium toruloids mating type a cells was purified to a single band on SDS-PAGE from cell-free extract by acid precipitation, ion exchange chromatogaphy andgel filtration. The determined molecular weight of he purified enzyme was about 95,000 by gel filtration and 100,000 daltons on SDS-polyacryamide gel electrophoresis. This enzyme didn't show any activity change by several metal ions except 15.4% decrease by {TEX}$Mn^{2+}${/TEX} and was strongly inhibited by 2-mercaptoethanol and SDS. The invertase maintained its activity at high level until 70℃, but inactivated at 80℃ almost completely. The optimal temperature and pH of the enzyme were about 60℃ and pH 5.0, respectively. The stable pH range of invertase was narrow from pH 3.0 to 6.0. The Km value and isoelectric point of enzyme were 3.4×{TEX}$10^{3}${/TEX} M, pH 4.4, respectively.

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