• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1630-1636
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• 2010

During the fermentation process of Saccharomyces cerevisiae, yeast cells must rapidly respond to a wide variety of external stresses in order to survive the constantly changing environment, including ethanol stress. The accumulation of ethanol can severely inhibit cell growth activity and productivity. Thus, the response to changing ethanol concentrations is one of the most important stress reactions in S. cerevisiae and worthy of thorough investigation. Therefore, this study examined the relationship between ethanol tolerance in S. cerevisiae and a unique protein called alcohol sensitive RING/PHD finger 1 protein (Asr1p). A real-time PCR showed that upon exposure to 8% ethanol, the expression of Asr1 was continuously enhanced, reaching a peak 2 h after stimulation. This result was confirmed by monitoring the fluorescence levels using a strain with a green fluorescent protein tagged to the C-terminal of Asr1p. The fluorescent microscopy also revealed a change in the subcellular localization before and after stimulation. Furthermore, the disruption of the Asr1 gene resulted in hypersensitivity on the medium containing ethanol, when compared with the wild-type strain. Thus, when taken together, the present results suggest that Asr1 is involved in the response to ethanol stress in the yeast S. cerevisiae.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.395-404
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• 2017

Fungal cell walls and cell membranes are the main targets of antifungals. In this study, we report on the antifungal activity of an ethanol extract from Paeonia lactiflora against Candida albicans, showing that the antifungal activity is associated with the synergistic actions of preventing cell wall synthesis, enabling membrane depolarization, and compromising permeability. First, it was shown that the ethanol extract from P. lactiflora was involved in damaging the integrity of cell walls in C. albicans. In isotonic media, cell bursts of C. albicans by the P. lactiflora ethanol extract could be restored, and the minimum inhibitory concentration (MIC) of the P. lactiflora ethanol extract against C. albicans cells increased 4-fold. In addition, synthesis of $(1,3)-{\beta}-{\small{D}}-glucan$ polymer was inhibited by 87% and 83% following treatment of C. albicans microsomes with the P. lactiflora ethanol extract at their $1{\times}MIC$ and $2{\times}MIC$, respectively. Second, the ethanol extract from P. lactiflora influenced the function of C. albicans cell membranes. C. albicans cells treated with the P. lactiflora ethanol extract formed red aggregates by staining with a membrane-impermeable dye, propidium iodide. Membrane depolarization manifested as increased fluorescence intensity by staining P. lactiflora-treated C. albicans cells with a membrane-potential marker, $DiBAC_4(3)$ ((bis-1,3-dibutylbarbituric acid) trimethine oxonol). Membrane permeability was assessed by crystal violet assay, and C. albicans cells treated with the P. lactiflora ethanol extract exhibited significant uptake of crystal violet in a concentration-dependent manner. The findings suggest that P. lactiflora ethanol extract is a viable and effective candidate for the development of new antifungal agents to treat Candida-associated diseases.

• 한국식품저장유통학회:학술대회논문집
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• 한국식품저장유통학회 2003년도 제23차 추계총회 및 국제학술심포지움
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• pp.200.2-201
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• 2003

본 연구에서는 생약재를 사용하여 부패에 관여하는 5종의 곰팡이(Aspergillus niger, Mucor miehei, Penicillium rugullosum, Aspergillus oryzae, Trichoderma reesei)에 대한 항 곰팡이 활성을 탐색하였다. 생약재의 물 추출물과 70% ethanol 추출물을 사용하여 5종의 곰팡이에 대한 항 곰팡이 활성을 측정한 결과, 물 추출물의 경우 갈근 추출물과 황련추출물에서 항 곰팡이 활성이 우수한 것으로 나타났다. 갈근 물 추출물은 A. niger, M. miehei, P. rugullosum의 3가지 균종에 대해서 모두 높은 항 곰팡이 활성을 나타냈으며, 황련 물 추출물은 M. miehei의 균종에 대해서 높은 항 곰팡이 활성을 나타내었다. 70% ethanol 추출물의 항 곰팡이 실험 결과, 물 추출물보다 좋은 항 곰팡이 활성을 나타내었다. 특히, 계피의 70% ethanol 추출물에서는 A. niger, M. miehei, P. rugullosum, A. oryzae, T reesei의 5가지 균종에서 모두 우수한 항 곰팡이 활성을 나타냈으며, 이 외에도 감초의 70% ethanol 추출물에서는 A. niger균주에 대해, 육두구와 호장근 및 황련의 70% ethanol 추출물에서는 M miehei 균주에 대해 우수한 항 곰팡이 활성을 나타냈으며, 또한 초두구의 70% ethanol추출물에서는 T reesei 균주에 대해 좋은 항 곰팡이 활성을 나타내었다. 항 곰팡이 효능이 우수한 생약재를 선별하고 이들 생약재로부터 추출한 다양한 획분을 사용하여 농도별로 항 곰팡이 활성을 조사한 결과, 갈근의 물 추출물의 경우에는 4가지 균종에서 모두 높은 항균활성을 나타냈으며, 낮은 농도에서도 높은 항 곰팡이 활성을 나타내었다. 이 외에도 황련의 물 추출물은 M. miehei 균주에서 우수한 항 곰팡이 활성을 나타내었다. 생약재의 70% Ethanol추출물을 농도별로 제조하여 항 곰팡이 활성을 살펴 본 결과, 70% ethanol추출물에서는 계피와 파고지, 초두구, 황련이 항 곰팡이 활성이 우수하였으며, 특히 계피의 70% ethanol 추출물에서는 5가지 균종에서 모두 우수한 항 곰팡이 활성이 나타났다.

• Archives of Pharmacal Research
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• 제19권5호
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• pp.374-380
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• 1996

Sprague-Dawley male rats (150 g) were chronically treated with 5 v/v % ethanol admixed with nutritionally complete liquid diet and fed ad libitum for 3 weeks. One half of each group was exposed to cold stress at 4 ^{\circ}C either for 24 h (for determination of mRNA by in situ hybridization) or for 48 h (for determination of enzyme activity). Chronic ethanol treatment (ethanol) did not affect tyrosine hydroxylase(TH) mRNA level in locus coeruleus(LC) of brain and adrenal medulla(AM) compared to controls. Cold stress showed strong increase of TH mRNA level in LC and AM compared to controls. Pretreated ethanol reduced the increased TH mRNA level by cold stress in LC and AM. Ethanol did not affect TH activity in LC and adenal glands(adrenals). Cold stress increased TH activity in LC but not in adrenals. Pretreated ethanol did not reduce the increased TH activity by cold stress in LC but this result was not shown in adrenals. Phenylethanolamine-N-methyltransferase(PNMT) activity in $C_{1}$$C_{2}$ and adrenals increased only in ethanol treated group. THese results suggest that ethanol does not affect TH mRNA level and activity in LC and adrenals, but increases PNMT activity in $C_{1}$$C_{2}$ and adrenals in normal rat. It is also suggested that pretreated ethanol reduces the magnitude of cold stress response, that is induction of TH mRNA in LC and AM, and does not reduce the protein activation of TH that is also cold stress response in LC.

• Korean Chemical Engineering Research
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• 제46권5호
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• pp.858-862
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• 2008

분리 당화발효(Separate Hydrolysis and Fermentation, SHF)는 당화와 발효공정을 따로 수행하는 방법으로 최종 생성물인 글루코오스에 의해 억제 영향을 받기 때문에 반응에 진행됨에 따라 축적된 글루코오스의 농도가 높아지면 반응이 종결되는 단점이 있다. 이를 극복하기 위해 효소의 양을 늘리는 방법이 있지만, 효소의 생산비용이 비싸기 때문에 경제적인 방법이 될 수 없다. 이러한 분리 당화발효 공정의 단점을 극복하기 위해서 동시당화발효 공정(Simultaneous Saccharification and Fermentation, SSF)은 하나의 반응기에서 당화와 발효를 동시에 수행한다. 동시당화발효 공정에서는 당화과정에서 글루코오스가 생성되자마자 효모가 발효과정을 통해 글루코오스를 바로 제거하기 때문에 반응기내에서 당의 축적을 최소화할 수 있다. 따라서 동시당화발효 공정은 최종 생성물의 억제 작용을 방지할 수 있고, 효소의 가수분해 반응을 향상시킬 수 있다. 본 연구에서는 동시당화발효에서 에탄올의 수율에 관여하는 조건들(pH, 반응온도, 효소 투입량, 반응시간)의 최적 조건을 찾는 연구를 수행하였다. 기질로는 감자전분을 사용하였고, 효소는 glucoamylase, 균주는 Saccharomyces cerevisiae가 각각 사용되었다. 동시당화발효의 최적의 조건은 pH 4, 온도 38로 나타났다. 최적의 조건으로 감자전분을 동시당화발효하였을 때 반응 18시간 후에 에탄올은 최대 수율 86%에 도달하였다.

• Journal of Ginseng Research
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• 제12권1호
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• pp.76-86
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• 1988

쥐에게 물대신 12% 에탄올(대조군) 또는 인삼사포닌 분획을 포함한 12% 에탄올(시험군)을 6일간 투여한 후 간과 혈청에서의 acetaldehyde 농도와 간의 [$NAD^+$]/ [NADH] 및 [$NADP^+$]/[NADPH] 비율을 조사하였다. 대조군의 간과 혈청의 acetaldehyde 농도는 물로 사육한 정상군에 비해 훨씬 높았으나 물대신 인삼사포닌을 포함한 에탄올을 투여한 시험군의 경우는 정상군에 비해 약간 높았을 뿐이었으며 [$NAD^+$]/ [NADH] 비의 감소율도 대조군보다는 시험군이 훨씬 작았다. l-$^{l4}C$]-ethanol을 함유한 10% ethanol을 1ml 복강으로 투여하고 30분 후의 간의 지방질의 방사능을 분석한 결과 시험군의 간 지방질의 전체 방사능은 대조군보다 훨씬 낮았고 인산지방질, 콜레스테롤, 지방산, 중성지방과 같은 지방질의 분석결과는 에탄올 투여로 인한 인산지방질 생합성 저하와 지방산 및 중성지방의 생합성 증가현상이 인삼사포닌의 투여로 개선되는 것으로 관찰되었다.

• Bulletin of the Korean Chemical Society
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• 제34권8호
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• pp.2413-2418
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• 2013

Forensic and legal medicine require reliable data to indicate excessive alcohol consumption. Ethanol is oxidatively metabolized to acetate by alcohol dehydrogenase and non-oxidatively metabolized to ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanol, or fatty acid ethyl esters (FAEE). Oxidative metabolism is too rapid to provide biomarkers for the detection of ethanol ingestion. However, the non-oxidative metabolite EtG is a useful biomarker because it is stable, non-volatile, water soluble, highly sensitive, and is detected in body fluid, hair, and tissues. EtG analysis methods such as mass spectroscopy, chromatography, or enzyme-linked immunosorbent assay techniques are currently in use. We suggest that nuclear magnetic resonance (NMR) spectroscopy could be used to monitor ethanol intake. As with current conventional methods, NMR spectroscopy doesn't require complicated pretreatments or sample separation. This method has the advantages of short acquisition time, simple sample preparation, reproducibility, and accuracy. In addition, all proton-containing compounds can be detected. In this study, we performed $^1H$ NMR analyses of urine to monitor the ethanol and EtG. Urinary samples were collected over time from 5 male volunteers. We confirmed that ethanol and EtG signals could be detected with NMR spectroscopy. Ethanol signals increased immediately upon alcohol intake, but decreased sharply over time. In contrast, EtG signal increased and reached a maximum about 9 h later, after which the EtG signal decreased gradually and remained detectable after 20-25 h. Based on these results, we suggest that $^1H$ NMR spectroscopy may be used to identify ethanol non-oxidative metabolites without the need for sample pretreatment.

• Preventive Nutrition and Food Science
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• 제9권4호
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• pp.352-360
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• 2004

To investigate the antioxidative effects of an etbylacetate fraction extracted from the flowers of Chrysanthemum indicum L. (Chrysanthemi Flos) on the antioxidative system and lipid profiles of rats with ethanol induced hepatotoxicity. Sprague-Dawley rats weighing $100\~150$ g were divided into 5 groups: normal group (NOR), Chrysanthemi Flos EtOAC fraction (200 mg/kg) treated group (S1), $35\%$ etbanol (10 mL/kg) treated group (S2), Chrysanthemi Flos EtOAC fraction (200 mg/kg) and ethanol concomitantly treated group (S3) and Chrysanthemi Flos EtOAC fraction (400 mg/kg) and ethanol concomitantly treated group (S4), respectively. The antioxidative activity of each fraction was decreased in order of EtOAC, n-hexane, n-BuOH, water and chloroform. The growth rates and feed efficiency ratios were decreased by ethanol treatment, but were gradually restored to similar levels as in the NOR group by administering Chrysanthemi Flos EtOAC fraction. The whole blood concentrations of total cholesterol and LDL-cholesterol, and the activities of ALT and AST that were elevated by ethanol were significantly decreased in the Chrysanthemi Flos EtOAC fraction treated groups. It was also observed that the activities of SOD, catalase, xanthine oxidase and GSH-Px elevated by ethanol in rat liver were markedly decreased in the Chrysanthemi Flos EtOAC fraction treated group as compared to S2. These results suggest that Chrysanthemi Flos EtOAC fraction has possible protective effects against ethanol induced hepatotoxicity in rat liver.

• KSBB Journal
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• 제23권6호
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• pp.504-508
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• 2008

국내산 라이밀을 이용한 바이오에탄올 생산을 위해 저온 전처리 공정을 도입하여 에탄올 생산성을 비교하였다. 라이밀의 경우 원료 특성상 증자 공정에서 점도 문제가 발생하는데, 이를 해결하기 위해 최적 전처리 조건을 탐색하였으며 이에 따른 에탄올 생산성을 비교하였다. 저온 조건과 점도 저하 효소를 사용함으로서 점도에 따른 발효 저해 현상 해결하였고 전처리 공정에 소요되는 전처리 공정비를 절감할 수 있었다. 또한 pH 조절(pH 4.5) 후 살균 처리 없이 바로 발효가 가능함을 확인할 수 있었다. 발효 초기 총당 함량은 $48{\pm}2.0\;g/L$이었으며, 발효 72시간 이후 에탄올 생성 농도는 $67.4{\pm}1.4\;g/L$, 톤당 에탄올 생산량은 410.9 L (dry base)로 효소 무첨가구보다 에탄올 농도와 톤당 수득량이 각각 15%, 20% 이상 증가하였다. 이와 같은 결과는 기존의 에탄올생산 공정과 비교하여 전처리 공정에 소요되는 시간을 30-50% 이상 줄일 수 있으며, 저온 공정에 따른 에너지 사용 절감 및 초기 시설 투자비를 줄일 수 있어 바이오에탄올 생산을 위한 대체 원료로 충분한 가능성을 보여 주었다.

• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.184-189
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• 2012

We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.