• Biotechnology and Bioprocess Engineering:BBE
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• 제9권1호
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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.

• 한국균학회지
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• 제43권3호
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• pp.196-199
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• 2015

본 연구에서는 국내 전통누룩에서 분리한 증류식 소주용 효모의 생리적 특성을 조사하고 동정하였다. 분리균(N4)의 26S rDNA-D1/D2 염기배열은 Saccharomyces cerevisiae 표준균주(NRRL Y-12632)와 100% 상동성을 나타내었고 다극출아에 의해서 증식하였다. Glucose, galactose, maltose, sucrose를 발효할 수 있으며 trehalose, raffinose, methylglucoside를 자화할 수 있었다. $40^{\circ}C$ 이상에서는 증식하지 못했으며 50% glucose와 10% NaCl 농도에서 성장할 수 있었다. 염기배열, 형태관찰 및 생리적 특성을 종합하여 분리균을 S. cerevisiae로 동정하였다.

• 한국식품영양과학회지
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• 제43권8호
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• pp.1174-1180
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• 2014

대추와 5개의 균주로 발효시킨 발효대추의 total polyphenol 함량과 in vitro에서의 radical 소거능을 검토한 결과 대추에 비해 발효대추에서의 total polyphenol 함량이 높게 나타났고 DPPH와 OH의 소거능이 우수하였으며, 특히 Saccharomyces cerevisiae로 발효시킨 대추가 가장 우수한 효과를 보였다. 이에 대추 추출물과 Sac. cerevisiae로 발효시킨 대추 추출물을 이용하여 C57BL/6N 탈모 마우스실험 모델 하에서 발모 성장 촉진 효과 및 탈모 예방 효과를 검토하였다. 제모 후 EtOH 도포군에서는 거의 발모가 진행되지 않았으나 Zizyphus jujuba(Zj) 도포군과 Sac. cerevisiae-fermented Zj(Zj-Y) 도포군에서는 전체적으로 발모가 진행되었고, 모발의 성장 속도는 minoxidil 도포군>Zj-Y 도포군> Zj 도포군> EtOH 도포군의 순으로 나타났다. 또한 자발적인 탈모에 대한 예방 효과도 대추, 특히 발효대추 도포군에서 우수함을 관찰할 수 있었다. 본 연구 결과로 대추에 비해 발효대추, 특히 Sac. cerevisiae로 발효시킨 대추가 항산화 효능이 뛰어나며, 발모 촉진 효과 및 탈모예방 효과가 우수함을 알 수 있었다.

• BMB Reports
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• 제31권2호
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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.

• 한국식품저장유통학회지
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• 제23권7호
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• pp.1042-1049
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• 2016

본 연구는 무독화 옻 추출물을 첨가하여 효모 종류별로 제조한 옻식초의 품질 특성을 비교하였다. Saccharomyces cerevisiae Fermivin과 non-Saccharomyces 속인 Pichia, Hanseniaspora 속 등 6종의 효모를 이용하여 알코올 발효시킨 결과, Fermivin이 알코올 함량이 가장 높았으며, Wickerhamomyces와 Pichia에서도 11.2, 12.2%의 알코올 함량을 나타내었다. 알코올 발효액에 초산균 A. pasteurianus KACC16934를 접종하여 초산발효 시킨 결과 pH와 적정 산도는 효모에 따른 차이가 있었다. W. anomalus N43-8을 제외한 모든 옻식초에서 발효 10일 후 pH 3.0 이하로 나타났으며, 적정산도도 W. anomalus N43-8로 발효한 식초가 5.17%로 가장 낮게 나타났고 C. tropicalis Y447로 발효한 식초는 6.66%로 가장 높은 산도를 보였다. 옻식초의 주요 유기산으로 acetic acid가 검출되었으며 W. anomalus N43-8 식초의 유기산은 35.88 mg/mL로 가장 낮았고 C. tropicalis Y447 식초는 55.49 mg/mL로 높은 함량을 나타내었다. 유리 아미노산은 모든 시험구에서 valine, leucine 등의 필수아미노산 7종이 검출되었으며 glutamic acid, alanine이 주요 아미노산으로 확인되었다. 유리아미노산은 $1,314.89{\sim}2,411.65{\mu}g/mL$로 효모의 종류에 따라 차이가 나타났다. 옻식초의 향기 패턴 분석 결과 H. opuntiae HP1-2와 P. kluyveri Frootzen이 비슷한 패턴을 보였으며, S. cerevisiae Fermivin과 C. tropicalis Y447, P. kudriavzeri N77-4가 비슷한 패턴을 보였다. 두 그룹과 W. anomalus N43-8은 전자코 분석 결과 각각의 뚜렷한 향기 패턴을 보였다. 본 연구를 발효 특성이 다른 효모로 식초를 제조할 때, 각각의 효모에 따라 이화학적 특성에 차이가 나는 것으로 보여진다.

• 한국식품영양학회지
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• 제13권2호
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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.

• 대한본초학회지
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• 제29권4호
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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.

• Biodesign
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• 제5권4호
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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.

• KSBB Journal
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• 제10권3호
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• pp.343-348
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• 1995

LeI은 스테로이드를 통울에 투여하였을 때 분비가 촉진되어 항염증성 효과를 나타내는 calcium 의 존성 phospholipid 결합 단백질이다. S. cerevisiae는 대장균과 통물세포의 장점을 모두 가지고 있으므로 동물세포 유래의 이종 단백질의 분비 생산에 많이 이용되고 있다. 본 연구에서는 GAL10 promoter­p ppL-LCI유전자 LCI terminator로 구성된 pYGLPT5 로 LCI을 S. cerevisiae SEY2102에서 발현 분비시키고 각 분획으로 나누어 LCI양을 비교한 결과 protoplast 68.6 %. periplasmic 24 %, culture supernatant 7.4%로 분포하였다. pYGLPT5로 형질전환된 S. cereviswe 2102를 유가 배양한 결과, 최종적인 LCI의 생산량은 약 $500mg/\ell$ 였다. LCI은 N 말단 부근에 $CA^{++}$ 결합부위가 있으므로 이를 이용하여 hydroxylapatite column chromatography로 정 제하 였다. 배지로 분비된 34kDa LCI을 ultrafiltration 과 hydroxylapatite column chromatography 의 두 단계로 순도 99% 이상으로 정제할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제23권10호
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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.