• Title/Summary/Keyword: Saccharomyces cerevisiae N4

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Mobile Phase Compositions for Ceramide III by Normal Phase High Performance Liquid Chromatography

  • Hong, Seung-Pyo;Lee, Chong-Ho;Kim, Se-Kyung;Yun, Hyun-Shik;Lee, Jung-Heon;Row, Kyung-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.1
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    • pp.47-51
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    • 2004
  • Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-hexane/ethanol, the first mobile phase composition, 95/5(v/v), was step-increased to 72/23(v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8(v/v/v), was step-increased to 75/10/15(v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

Characterization of Yeast for Soju (Distilled Spirits) from Korean Traditional Nuruk (국내 전통누룩에서 분리한 증류식 소주용 효모의 특성)

  • Choi, Han-Seok;Kang, Ji-Eun;Jeong, Seok-Tae;Kim, Chan-Woo;Takamine, Kazunori
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.196-199
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    • 2015
  • The occurrence and characterization of yeast isolated from Korean traditional nuruk were investigated. Sequences of the D1/D2 domain of the 26S rDNA were identical for a strain examined and had a similarity value of 100% compared to sequences of the type strain of Saccharomyces cerevisiae (NRRL Y-12632). The isolate able to assimilate with trehalose, raffinose, and methyl-glucoside assimilate was also capable of glucose, galactose, maltose, and sucrose fermentation. It did not proliferate at $40^{\circ}C$ or above, but was able to grow in concentration of 50% glucose and 10% NaCl. By combining nucleotide sequence, morphological observation, and physiological characteristics, the isolate was identified as S. cerevisiae.

Protective Effects of Zizyphus jujuba and Fermented Zizyphus jujuba from Free Radicals and Hair Loss (대추 및 발효대추의 라디칼 소거능 및 모발 성장 촉진 효과)

  • Jung, Ji Eun;Cho, Eun Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1174-1180
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    • 2014
  • This study investigated the antioxidative and hair growth-promoting activities of Zizyphus jujuba (Zj) and fermented Zj with Aspergillus oryzae, Bacillus subtilis, Bifidobacterium breve, Lactobacillus acidophilus, and Saccharomyces cerevisiae (Sac. cerevisiae). Among Zj and fermented Zjs, Sac. cerevisiae-fermented Zj (Zj-Y) exerted stronger scavenging activity against 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals than others. In addition, total polyphenol content of Zj-Y was higher than that of non-fermented Zj and other fermented Zj. This result indicates that fermentation of Zj by Sac. cerevisiae elevated antioxidative activity. Furthermore, using an alopecia model in C57B/6N mice, the hair growth activities of Zj and Zj-Y were investigated. The test samples, EtOH, minoxidil (MXD), Zj, and Zj-Y, were topically treated with 0.2 mL/day for 4 weeks. The experiments involved macroscopic observation and measurement of hair length methods. The results show that regrowth speed of hair was in decreasing order of MXD> Zj-Y> Zj> EtOH. The topical application of MXD and Zj-Y in mice promoted hair regrowth and prevented hair loss compared to the control group. The present study indicates that Zj-Y is a promising treatment for alopecia.

Expression and Secretion of Foreign Proteins in Yeast Using the ADH1 Promoter and 97 K Killer Toxin Signal Sequence

  • Hong, Seok-Jong;Kang, Hyen-Sam
    • BMB Reports
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    • v.31 no.2
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    • pp.123-129
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    • 1998
  • Foreign proteins, $endo-{\beta}-1,4-glucanase$ of Bacillus subtilis, preS1+S2 region of hepatitis B virus large surface antigen, human ${\beta}_2-adrenergic$ receptor ($h{\beta}_{2}AR$), and bovine growth hormone (bGH) were expressed in Saccharomyces cerevisiae and secreted into the medium. These proteins were expressed using the alcohol dehydrogenase I (ADH1) promoter of Saccharomyces cerevisiae and secreted by signal sequence of the 97 K killer toxin gene of doublestranded linear DNA plasmid (pGKL1) of S. cerevisiae. All these proteins underwent severe modifications; in particular, N-glycosylation in the case of $endo-{\beta}-1,4-glucanase$, $h{\beta}_2AR$, and preS1+S2. Seventy four percent of the expressed $endo-{\beta}-1,4-glucanase$ was secreted into the culture medium. Highly modified proteins were detected in the culture medium and in the cell. Expressed $h{\beta}_2AR$, which has seven transmembrane domains, remained in the cell. The degrees of secretion and modification and the states of proteins in the culture medium and in the cell were quite different. These results indicated that the nature of the protein has a critical role in its secretion and modifications.

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Characterization of acetic acid fermentation of detoxified Rhus verniciflua vinegar produced by various yeast strains (효모 종류에 따른 무독화 옻식초의 초산발효 특성)

  • Kim, Ji-Seon;Yeo, Soo-Hwan;Mun, Ji-Young;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1042-1049
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    • 2016
  • This study was conducted to investigate and compare the characteristics of acetic acid fermentation in detoxified Rhus verniciflua vinegar (DRV) produced by different yeast strains. The DRVs were prepared by static acetic acid fermentation using six different yeast strains (Saccharomyces cerevisiae Fermivin, Pichia kudriavzerii N77-4, Hanseniaspora pountiae HP1-2, Candida tropicalis Y447, Wickerhamomyces anomalus N43-8, and Pichia kluyveri Frootzen). Alcohol content of the S. cerevisiae Fermivin fermented DRV was highest 16.07%. Among the yeast strain DRVs, there were significant differences in alcohol content, but all alcohol levels were 11%. Moreover, there were differences in pH and titratable acidity of the DRVs. The organic acid content of the DRVs ranged from 35.88 to 55.49 mg/mL and there were significant differences among the yeast strain DRVs. Essential free amino acids, particularly glutamic acid, alanine, leucine and valine, were detected in each of the 6 DRVs. Electronic nose analysis revealed that three different volatile chemical patterns were present in the 6 DRVs. The results indicate that yeast strains with different characteristics can produce vinegars with different characteristics.

Production of Killer Toxin from a Mutant of Hansenular capsulata S-13 (Hansenular capsulata S-13의 변이주에 의한 Killer Toxin의생산)

  • 김재호;김나미;이종수
    • The Korean Journal of Food And Nutrition
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    • v.13 no.2
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    • pp.158-163
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    • 2000
  • Killer yeast, Hansenular capsulata S-13 were treated with heat, ethylmethane sulfonate and N-methyl-n'-nitro-n-nitrosoguanidine and a mutant(S13-E1), showing 2-fold higher killer toxin activity than that of parent strain to killer sensitive strain, Saccharomyces cerevisiae ATCC 38026 was obtained. Hansenular capsulata S13-E1 showed strong killer toxin activity to Saccharmyces mellis and Saccharomyces sal년 and four strains of gas-producing yeasts from traditional Doenjang and Kochujang. The culture condition for killer toxin production by Hansenular capsulata S13-E1 was optimized to be 1.0% potato extract, each 0.5% of peptone and glucose, and 0.025% MgSO4 with initial pH 4.5 at 3$0^{\circ}C$ and 36 hr of batch cultivation.

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Antibacterial activity of Bio-fermented Galla Rhios Extract (오배자 발효추출물의 항세균활성)

  • Doh, Eun Soo;Yoo, Ji Hyun
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.21-27
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    • 2014
  • Objectives : This experimental study was performed in order to investigate the antibacterial effect of bio-fermented Galla Rhois extract. Methods : The Galla Rhois extract was fermented by Streptococcus thermophilus, Saccharomyces cerevisiae and Lactobacillus delbrueckii, and their products was tested for antibacterial activity against six pathogenic microorganisms namely, Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Vibrio parahaemolyticus, Escherichia coli and Salmonella typhimurium by paper disc diffusion method. Results : The Galla Rhois fermented extract by Lactobacillus delbrueckii and Saccharomyces cerevisiae showed more effective antibacterial activity than not fermented extract against Bacillus subtilis and Vibrio parahaemolyticus. Antibacterial activity of fermented extract using especially Lactobacillus delbrueckii and Saccharomyces cerevisiae was proved that it was good with even 2 percents concentration. Antibacterial activity of Galla Rhois extract within pH 3 to pH 7 had been safe regardless of pH but low over pH 9. The growth of Bacillus cereus, Staphylococcus aureus, and Vibrio parahaemolyticus had a tendency to decrease depend on the increasing concentration of the extract. EtOEt, EtOAc and n-BuOH fractions of the Galla Rhois extract had a high level of antibacterial activity against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Vibrio parahaemolyticus, respectively. Surprisingly, EtOAc fractions of the Galla Rhois extract showed higher antibacterial activity against Vibrio parahaemolyticus alone. And antibacterial activity against six pathogenic microorganisms had a tendency to increase depend on the increasing concentration of the fractions of the Galla Rhois extract. Conclusions : Bio-fermented Galla Rhois extract, efficiently inhibited the growth of Bacillus cereus and Vibrio parahaemolyticus.

Crystallization and X-ray crystallographic analysis of the PH-like domain of lipid transfer protein anchored at membrane contact sites from Saccharomyces cerevisiae

  • Tong, Junsen;Im, Young Jun
    • Biodesign
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    • v.5 no.4
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    • pp.136-140
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    • 2017
  • Lam6 is a member of sterol-specific ${\underline{l}ipid$ transfer proteins ${\underline{a}}nchored$ at ${\underline{m}ebrane$ contact sites (LAMs). Lam6 localizes to the ER-mitochondria contact sites by its PH-like domain and the C-terminal transmembrane helix. Here, we purified and crystallized the Lam6 PH-like domain from Saccharomyces cerevisiae. To aid crystallization of the Lam6 PH-like domain, T4 lysozyme was fused to the N-terminus of the Lam6 PH-like domain with a short dipeptide linker, GlySer. The fusion protein was crystallized under the condition of 0.1 M HEPES-HCl pH 7.0, 10% (w/v) PEG 8000, and 0.1 M $Na_3$ Citrate at 293K. X-ray diffraction data of the crystals were collected to $2.4{\AA}$ resolution using synchrotron radiation. The crystals belong to the orthorhombic space group $P2_12_12_1$ with unit cell parameters $a=59.5{\AA}$, $b=60.1{\AA}$, and $c=105.6{\AA}$. The asymmetric unit contains one T4L-Lam6 molecule with a solvent content of 58.7%. The initial attempt to solve the structure by molecular replacement using the T4 lysozyme structure was successful.

Production and Purification of Human Lipocortin-I Secreted by Recombinant Saccharomyces cerevisiae (재조합 Saccharomyces cerevisiae로부터 인체 리포코틴-I의 분비 생산 및 정제)

  • 김병문;정봉현
    • KSBB Journal
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    • v.10 no.3
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    • pp.343-348
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    • 1995
  • Human lipocorin-I(LCI) is a calcium ion-dependent and phospholipid-binding protein which exhibits an anti-inflammatory activity by inhibiting phospholipase A2 activity. In this study, the LCI gene containing its own terminator region was joined to GAL10 promoter-ppL (prepro-leader sequence of mating factor a). An ATG start codon of LCI gene was placed at downstream with KR endoprotease recognition site(Lys-Arg) of ppL. Recombinant S. cerevisiae harboring the LCI expression/secretion vector, pYGLPT5, was aerobicall grown on a liquid YPDG medium al $30^{\circ}C$ for 72hys. The whole cell and culture supernatant were separated after centrifugation, and the expressed LCI was analyzed by SDS-PAGE and western blotting methods. A majority fraction of the expressed LCI was found to be accumulated in the intracellular fraction, resulting in very low secretion efficiency of about 7.4%. About $500mg/\ell$ of LCI was extracellularly produced by the fed-batch culture employing the controlledfeeding of glucose and galactose. The secreted LCI was purified by ultrafiltration and hydroxylapatite column chromatography, and a purity of more than 99% was obtained.

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Development of a Bioconversion System Using Saccharomyces cerevisiae Reductase YOR120W and Bacillus subtilis Glucose Dehydrogenase for Chiral Alcohol Synthesis

  • Yoon, Shin Ah;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1395-1402
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    • 2013
  • Reductases convert some achiral ketone compounds into chiral alcohols, which are important materials for the synthesis of chiral drugs. The Saccharomyces cerevisiae reductase YOR120W converts ethyl-4-chloro-3-oxobutanoate (ECOB) enantioselectively into (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB), an intermediate of a pharmaceutical. As YOR120W requires NADPH as a cofactor for the reduction reaction, a cofactor recycling system using Bacillus subtilis glucose dehydrogenase was employed. Using this coupling reaction system, 100 mM ECOB was converted to (R)-ECHB. A homology modeling and site-directed mutagenesis experiment were performed to determine the NADPH-binding site of YOR120W. Four residues (Q29, K264, N267, and R270) were suggested by homology and docking modeling to interact directly with 2'-phosphate of NADPH. Among them, two positively charged residues (K264 and R270) were experimentally demonstrated to be necessary for NADPH 2'-phosphate binding. A mutant enzyme (Q29E) showed an enhanced enantiomeric excess value compared with that of the wild-type enzyme.