• Title/Summary/Keyword: S cerevisiae

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Studies on the wild yeasts in Korea (II) (한국산 야생효모에 관한 연구 2)

  • 박명삼;라철호
    • Korean Journal of Microbiology
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    • v.8 no.3
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    • pp.95-102
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    • 1970
  • From the crops Drosophila collected in Mt. Sokni and Mt.Kyeryong, 7 strains were isolated and then 6 species of wild yeast were identified. 1) Of these six species of wild yeasts two were to be of genus Saccharomyces(Ascosporgenous), two Torulopsis and two Trichosporon (both genuses of Asporogenous). 2) It was found that the fermentation of the wild yeasts isolated from Drosophila was much better than that of any others ; in particular, S. florentinus and S. cerevisiae were good in fermenting maltose. 3) After being cultivated in malt extract agar medium at $25^{\circ}C$ for 3 days, the vegetative cells were found to be big but Torulopsis cells small. 4) It was also observed that the species of yeasts used fro food by Drosophila largely depends on genus and species of Drophila. 5) Of the yeasts isolated from the Drosophila, Trichosporon capitatum and Torulopsis dattila, which has not previously been recorded, were identified. 6) It is believed, therfore, that S.florentinus, powerful in fermenting maltose, will be extremely useful in terms of industrial application.

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Isolation of the Phosphoribosyl Anthranilate Isomerase Gene (TRP1) from Starch-Utilizing Yeast Saccharomycopsis fibuligera

  • Park, Eun-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1324-1327
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    • 2015
  • The nucleotide sequence of the TRP1 gene encoding phosphoribosyl anthranilate isomerase in yeast Saccharomycopsis fibuligera was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 759 bp, including the stop codon, encoding a 252 amino acid residue. The deduced amino acid sequence of Trp1 in S. fibuligera was 43.5% homologous to that of Komagataella pastoris. The cloned TRP1 gene (SfTRP1) complemented the trp1 mutation in Saccharomyces cerevisiae, suggesting that it encodes a functional TRP1 in S. fibuligera. A new auxotrophic marker to engineer starch-degrading yeast S. fibuligera is now available. The GenBank Accession No. for SfTRP1 is KR078268.

The MALDI-TOF MS determination of yeast proteins producing $H_2S$ (MALDI-TOF MS를 이용한 효모에서의 황화수소 생성 단백질의 동정)

  • Cho, Hyun-Nam;Fan, Lu-An;Yoo, Dong-Chan;Yang, Seun-Ah;Lee, In-Seon;Kim, Jae-Hyung;Baek, Hyo-Hyun;Jhee, Kwang-Hwan
    • KSBB Journal
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    • v.23 no.5
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    • pp.425-430
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    • 2008
  • Hydrogen sulfide ($H_2S$) is a by-product of metabolism of amino acids including sulfur and alcoholic fermentation, it is generally thought of in terms of a poisonous gas. Though $H_2S$ can have a negative impact on the perceived quality of fermented drinks due to an undesirable aroma, it plays prominent roles as a neuromodulator in the mammalian brain as well as a smooth muscle relaxant. Nowadays studies on the proteins which produce $H_2S$ are carried out in various fields such as structure, function, and metabolism. Here we propose to develop a simple and rapid $H_2S$ forming assay method, which will lead to speed up preparing the $H_2S$ forming proteins for identification by MALDI-TOF MS analysis. We detected three kinds of proteins which produce $H_2S$ in the crude extract of Saccharomyces cerevisiae. Those proteins were cystathionie $\beta$-synthase, O-acetylserine sulfhydrylase, and cystathionine $\gamma$-lyase.

Effects of Fermented Sparassis crispa Stipe Extract Supplemented Diet on the Immune Responses of Philippines Eel, Anguilla bicolor (꽃송이버섯 기부 발효물 첨가 사료가 장어의 면역반응에 미치는 영향)

  • Kim, Eun-Ju;Seo, Seung-Ho;Park, Seong-Eun;Kang, Min-Soo;Son, Hong-Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.10
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    • pp.1151-1157
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    • 2017
  • This study examined the immune response of Philippines eel (Anguilla bicolor) to the oral administration of fermented Sparassis crispa stipe extract for 6 weeks. The S. crispa extract fermented with Lactobacillus plantarum showed a higher total phenol content (301.68 ppm) and DPPH radical scavenging activity (63.9%) than those fermented with other strains. Therefore, L. plantarum was selected as a suitable starter culture for the fermentation of S. crispa stipe. The eels were fed a commercial diet supplemented with 1% of fermented S. crispa stipe extract for 6 weeks. The mortality rate of the eels fed the supplemented diet was significantly lower than those of the control after 6 weeks. The lysozyme activity of the serum was increased significantly (12.33 ${\rightarrow}$ 54.66 units) after 6 weeks in the eel fed supplemented diets of fermented S. crispa stipe. The serum of the eel fed the supplemented diet of the S. crispa stipe extract showed higher bactericidal activity. These results suggest that both the S. crispa stipe extract and fermented S. crispa stipe have strong potential to activate the innate immune response of the Philippines eel.

Fermentable and Nutritional Characteristics of Brewery Meal-Based Fermented Feedstuffs Supplemented with Aspergillus Oryzae and Saccharomyces Cerevisiae (Aspergillus Oryzae 및 Saccharomyces Cerevisiae를 첨가하여 제조한 맥주박 위주 발효사료의 발효 특성 및 영양학적 특성)

  • Shin, Jong-Suh;Park, Byung-Ki;Kim, Byong-Wan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.4
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    • pp.297-306
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    • 2005
  • This study was conducted to compare and evaluate fermentable and nutritional characteristics of brewery meal-based fermented feedstuffs supplemented with Aspergillus oryzae (AO) and Saccharomyces cerevisiae (SC). Experiments were divided into three treatment groups; fermented feedstuff supplemented with $1\%$ of AO(FFAO), fermented feedstuff supplemented with $1\%$ of SC(FFSC), and fermented feedstuff supplemented with $0.5\%$ of AO and $0.5\%$ of SC(FFAS). For changes of crude protein contents by 48 h fermentation, there were no significant differences among treatments. Ether extract(EE) contents were significantly increased by 48 h fermentation (p<0.05). Neutral detergent fiber(NDF) contents of FFAO, FFSC and FFAS were significantly decreased by 48 h fermentation(p<0.05), but acid detergent fiber(ADF) and acid detergent lignin (ADL) contents were not different. The pH of FFAO and FFAS was decreased more rapidly than that of FFSC(p<0.05), reaching a plateau after 24 h. Alcohol content was increased rapidly until 18 h in FFAO and was increased rapidly until 12 h in FFSC and FFAS, and alcohol content of FFAO, FFSC and FFAS was maintained constantly after 24 h. The ammonia N content of FFAO, FFSC and FFAS was 0.022, 0.073 and $0.040\%$ at 48 h, respectively, and then ammonia N was over twice higher in FFSC than in FFAO and FFAS(p<0.05). Dextrose content was increased until 6 h in FFAO but was rapidly decreased in FFSC and FFAS until 6 h(p<0.05). Lactate content was higher in FFAO and FFAS than in FFSC(p<0.05). Consequently, when we added AO in formulation of fermented feedstuff with brewery meal which moisture content was high, EE, alcohol, and lactate contents were increased, but NDF and ammonia N contents were reduced. Therefore, it is expected that AO will be effective to increase the feed value and the preservation of fermented feedstuff with a high moisture content.

Studies on the inhibitory substance of yeast growth - (Part 1) Isolation and biochemical characteristics of yeast growth inhibitor - (항효모성물질(抗酵母性物質)에 관(關)한 연구(硏究) - (제 1 보)(第 1 報) 분리(分離) 및 그 생화학적작용(生化學的作用)과 성질(性質)에 대(對)해서 -)

  • Seu, Jung-Hwn;Kho, Yung-Hee
    • Applied Biological Chemistry
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    • v.15 no.1
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    • pp.19-25
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    • 1972
  • A kind of peptide which posseses an yeaststatic activity was isolated from Astragalus membranaceus Bunge and following characteristics was obtained. 1. The isoelectric pH of this peptide was 8.2 and histidine, an alkaline amino acid, was identified from this peptide. 2. This substance showes conspicuous heat stability and does not indicate any remarkable reduction of yeaststatic activity even for 5 hours treatment at $100^{\circ}C$. or for 30 minutes at $121^{\circ}C$. 3. The inhibitory activity of the yeast growth is not originated from the yeastcidal action but yeaststatic effect of this sample. 4. The sample shows strong stability ranging from pH 2 to 10. 5. The saccharide; glucose, sucrose, maltose, gives no effect on the yeaststatic activity of the sample even high concentration, 15 percent, and also no effect gives by magnesium, calcium and phosphate salts. 6. The available concentration of this sample on the inhibition of yeast growth was located at the ppm extent, for example, the concentration of fifty percent growth inhibition to Saccharomyces cerevisiae or S. carsbergensis was 4 ppm and 3 ppm to Candida pulcherrima, 13 ppm to S. coreanus, 18 ppm to S. sake and 38 ppm to C. tropicalis. 7. On the alcohol fermentation of S. coreanus, the peptide, an yeast growth inhibitor, gives no effect at all. 8. This substance is named as Astradix-P (Astragalus membranaceus, Radix, Peptide).

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Construction of Starch-assimilating and Ethanol-fermenting Yeast by Protoplast Fusion (원형질 융합에 의한 전분으로부터 에탄올 발효효모균주의 개량)

  • 이혜정;이지나;천경숙;박소영;마은애;민경희
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.546-552
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    • 1992
  • Ethanol-tolerant strain, S. eerevisiae BUI a26 ($alc^r thr^-$) and gJucoamylase-producing strain, S diastatieus AI5a6 (STA+ hom-) were prepared by means of genetic manipulation, Protoplast fusion was carried out to introduce STA gene from AI5a6 strain to BUla26 strain, Protoplast formation was shown at 0,8 M sorbitol and 200 Jig/ml to 400 Jig/ml zymolyase treatment for 2 hours incubation, Fusion frequency was $ 3.25 {\times} 10^{-3}$ to the regenerated protoplast number using PEG 6000 for 90 min incubation. The excellent fusants with genotype of STA- $alc^r thr^-$ hom+/STA+ ($alc^s thr^+$ hom- (2n), F7 and FIO, were selected by ethanol-tolerant, ethanol fermentation, and glucoamylase production tests, Glucoamylase production of AI5a6 showed 2,7 units, but 4.2 or 8.4 units for F7 or FIO fusant at $30^{\circ}C$, Ethanol fermentation from 32% glucose by BUla26 was 14,0%(v/v) in fermentaion medium for 5 days incubation, but 14.5% or 15,0% for F7 or FIO strain, respectively. Ethanol fermentation from 5% starch was 2,0% by F7, or 1.8% by FIO strain in fermentation medium for 5 days fermentation.

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Culture Conditions for Glucoamylase Production and Ethanol Productivity of Heterologous Transformant of Saccharomyces cerevisiae by Glucoamylase Gene of Saccharomyces diastaticus (Transformant의 Glucoamylase 생성조건과 Ethanol 발효성)

  • Kim, Young-Ho;Jung-Hwn Seu
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.494-498
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    • 1988
  • The optimum conditions for glucoamylase production, and ethanol productivity of the transformant TSD-14 were investigated as compared with the parental strains. The properties of TSD-14 were comparatively similar to the donor S. diastaticus IFO 1046 as regards the conditions of glucoamylase production and ethanol productivity. The soluble starch was the most effective carbon source for the glucoamylase production. While inorganic nitrogen sources did not prompt cell growth and enzyme production, the organic nitrogen sources generally enhanced both cell growth and glucoamylase production. The metal salts such as FeSO$_4$, MgSO$_4$, MnCl$_2$, and NiSO$_4$were favorable to the enzyme production. And the optium temperature and initial pH for glucoamylase production were 3$0^{\circ}C$ and 5. The transformant TSD-14 produced 8.3%(v/v) ethanol from 15% sucrose medium, 4.8%(v/v) ethanol from 15% soluble starch medium, and 7.5%(v/v) ethanol from 15% liquefied potato starch medium. The corresponding fermentation efficiency were 84% , 45% and 70%, respectively.

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Transcriptional Regulation of a DNA Repair Gene in Saccharomyces cerevisiae

  • Jang, Yeon-Kyu;Sancar, Gwen-B.;Park, Sang-Dai
    • Proceedings of the Zoological Society Korea Conference
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    • 1998.10b
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    • pp.113-113
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    • 1998
  • In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

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Selection of Constitutive Promoter for Exoinulinase Production in Fed-Batch Culture of Recombinant Yeast (재조합 효모의 유가배양에서 Exoinulinase생산을 위한 Promoter의 선별)

  • 김이경;고지현;김연희;김성구;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.206-211
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    • 2001
  • In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

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