• 산업식품공학
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• 제15권1호
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• pp.22-27
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• 2011

본 연구에서는 강원도 춘천지역에서 발생하는 환경 오폐수를 시료로 하여 미생물을 혼합하여 제조한 수질정화제를 실제 하천과 호수의 오염환경과 유사한 배양기내에서 생물학적반응을 시켜오염물질의정화정도를측정하고평가하였다. 사용된 미생물은 식물성 유산균(vegetable Lactobacillus fermentum), 효모(Saccharomyces cerevisiae)와 고초균(Bacillus subtilis)이다. 두가지의미생물혼합제를투입하여정화정도를 실험하였다. 실제 하천에 투입하여 정화되는 과정과 유사하게 반응을 시키고자 반응 온도는 $21({\pm}3)^{\circ}C$로 하였으며 완전교반을 진행하였다. 사용한 시료의 오폐수의 오염도와 미생물에 의한 정화정도를 평가하기 위하여 총질소 함량과 총인 함량을 측정하였다. V.L.F.와 S.C.의 혼합물로 구성된 혼합제의 경우 총질소함량을 감소시키는데 매우 효과적임을 보여주었으며, V.L.F., S.C.,와 B.S.로 구성된 혼합제의 경우 그 총인의 함량을 저감화하는데 효과적임을 알 수 있었다. 본 실험에서 얻어진 실험 데이터를 통하여 V.L.F.와 S.C.의 혼합제가 오폐수의 정화반응에서 나타나는 반응상수값을 도출하였으며 그 값은 0.178 $day^{-1}$이었다. 본 연구의 결과를 바탕으로 저감화하고자 하는 성분에 따라 미생물의 종류와 그 혼합비를 조절하는 최적화과정이 필수적임을 알 수 있었으며 또한 곡선적합으로 도출한 반응상수값은 미생물을 이용하여 호수와 하천의 정화정도를 simulation함에 필요한 실험인자로서 응용이 가능하게 되었다.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.761-767
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• 2022

EHT1 and EEB1 are the key Saccharomyces cerevisiae genes involved in the synthesis of ethyl esters during wine fermentation. We constructed single (Δeht1, Δeeb1) and double (Δeht1Δeeb1) heterogenous mutant strains of the industrial diploid wine yeast EC1118 by disrupting one allele of EHT1 and/or EEB1. In addition, the aromatic profile of wine produced during fermentation of simulated grape juice by these mutant strains was also analyzed. The expression levels of EHT1 and/or EEB1 in the relevant mutants were less than 50% of the wild-type strain when grown in YPD medium and simulated grape juice medium. Compared to the wild-type strain, all mutants produced lower amounts of ethyl esters in the fermented grape juice and also resulted in distinct ethyl ester profiles. ATF2, a gene involved in acetate ester synthesis, was expressed at higher levels in the EEB1 downregulation mutants compared to the wild-type and Δeht1 strains during fermentation, which was consistent with the content of acetate esters. In addition, the production of higher alcohols was also markedly affected by the decrease in EEB1 levels. Compared to EHT1, EEB1 downregulation had a greater impact on the production of acetate esters and higher alcohols, suggesting that controlling EEB1 expression could be an effective means to regulate the content of these aromatic metabolites in wine. Taken together, the synthesis of ethyl esters can be decreased by deleting one allele of EHT1 and EEB1 in the diploid EC1118 strain, which may modify the ester profile of wine more subtly compared to the complete deletion of target genes.

• 한국식품저장유통학회지
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• 제16권6호
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• pp.977-984
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• 2009

Campbell Early 포도 품종은 생육 조건이 우리나라의 기후 조건에 매우 적합하며, 생과로서 향이 좋고, 오랜 기간 생과로 먹어 왔으므로 우리의 입맛에 잘 맞는 포도로서 과즙의 제조에는 적합한 특성을 가지고 있으나 당의 함량이 14-15%로서 포도주 가공을 위하여 보당을 하여야 하는 등 포도주 제조에는 다소 문제점이 있다. 국내산 포도주의 품질 개선을 위한 원료 전처리 방법으로 본 연구에서는 동결 농축 기술을 이용하여 Campbell Early 포도 과즙을 농축한 후 다양한 포도주 효모를 사용하여 무가당 포도주의 제조 시험을 행하였다. 포도주 효모로는 산업용 포도주 효모인 S. cerevisiae OC2, S. cerevisiae Fermivin, S. cerevisiae W-3를 이용하여 농축 포도 과즙의 알코올 발효를 행하면서 발효 특성과 발효 후 여과하여 제성한 포도주의 품질 특성을 조사하였다. 발효가 끝날 시점의 알코올 함량은 12.6-13.0%로 비슷하였으며 환원당의 함량은 거의 흔적 정도로 검출되었으며 pH는 약 3.6-3.7, 총산 함량은 0.6-0.7%로 모든 시험구에서 정상적으로 발효가 일어났음을 알 수 있었다. 발효 종료 후 여과한 포도주의 아세트알데히드, 메탄올, fusel oil의 함량은 식품공전에 명시된 과실주의 기준치보다 매우 낮게 나왔으며 총 폴리페놀 함량이 모든 시험구에서 1.32-1.67 mg/mL 정도를 나타내었다. 각 시험구의 포도주는 모두 농축으로 인하여 진한 색을 띄었고 특히 적색을 나타내는 520nm에서의 흡광도가 높았으며 이는 색도인 L, a, b 값을 통해서도 확인하였다. 관능 검사를 실시한 결과는 색은 S. cerevisiae W-3 포도주가 가장 우수한 것으로 나타났으며 향과 맛은 S. cerevisiae OC2 포도주에서 가장 우수한 점수를 받았으며 전반적인 기호는 S. cerevisiae Fermivin 포도주가 가장 우수한 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.215-221
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• 1994

Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.767-774
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• 2010

In this study, the problems of high caloric content, increased maturation time, and off-flavors in commercial beer manufacture arising from residual sugar, diacetyl, and acetaldehyde levels were addressed. A recombinant industrial brewing yeast strain (TQ1) was generated from T1 [Lipomyces starkeyi dextranase gene (LSD1) introduced, ${\alpha}$-acetohydroxyacid synthase gene (ILV2) disrupted] by introducing Saccharomyces cerevisiae glucoamylase (SGA1) and a strong promoter (PGK1), while disrupting the gene coding alcohol dehydrogenase (ADH2). The highest glucoamylase activity for TQ1 was 93.26 U/ml compared with host strain T1 (12.36 U/ml) and wild-type industrial yeast strain YSF5 (10.39 U/ml), respectively. European Brewery Convention (EBC) tube fermentation tests comparing the fermentation broths of TQ1 with T1 and YSF5 showed that the real extracts were reduced by 15.79% and 22.47%; the main residual maltotriose concentrations were reduced by 13.75% and 18.82%; the caloric contents were reduced by 27.18 and 35.39 calories per 12 oz. Owing to the disruption of the ADH2 gene in TQ1, the off-flavor acetaldehyde concentrations in the fermentation broth were 9.43% and 13.28%, respectively, lower than that of T1 and YSF5. No heterologous DNA sequences or drug resistance genes were introduced into TQ1. Hence, the gene manipulations in this work properly solved the addressed problems in commercial beer manufacture.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1539-1545
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• 2010

In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.390-399
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• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• 한국미생물·생명공학회지
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• 제40권3호
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• pp.220-225
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• 2012

실험실용 효모 CEN.PK2-1D와 산업용 효모 JHS100와 JHS200의 ethanol 발효성능을 순수배지와 카사바 당화액에서 비교하고 발효특성을 규명하기 위해 세포성장속도와 ethanol 내성, 부산물 glycerol과 acetate의 생성에 대해 비교하였다. JHS100과 JHS200은 CEN.PK2-1D과 비교하여 세포성장이 빨랐으며 ethanol에 대한 내성이 높았다. 순수YP 배지에 300 g/L glucose를 탄소원으로 이용한 회분식 배양에서 세 효모 모두 0.46 g/g의 ethanol 생산수율을 나타냈으며 생산성은 세포성장속도가 가장 빨랐던 JHS100이 3.05 g/L-hr로 가장 높아서 최종적으로 JHS100 균주는 136.6 g/L의 ethanol을 생산하였다. 카사바 당화액에 질소원을 추가하지 않은 배지를 이용한 혐기성 회분식 배양에서 산업용 효모 JHS100은 106.1 g/L ethanol을 최종적으로 생산하였고, 0.42 g/g 생산수율과 3.15 g/L-hr 생산성을 보였다. 특히, 카사바 당화액과 순수 YP배지를 이용한 발효의 ethanol 생산변수를 비교할 경우, 실험실용 효모 CEN.PK2-1D는 생산수율과 생산성이 각각 19%와 17% 감소한 반면, 산업용 효모 JHS100과 JHS200은 생산수율이 8% 감소하였고 유사한 생산성을 보였다. 또한, ethanol 생산과정의 최대 부산물인 glycerol과 acetate의 생산에 대해서 JHS100과 JHS200이 CEN.PK2-1D에 비하여 크게 낮았다. 따라서 산업용 효모인 JHS100과 JHS200의 뛰어난 ethanol 발효성능은 빠른 세포성장과 높은 ethanol 내성, 낮은 질소원 요구성, 부산물인 glycerol과 acetate의 낮은 생산성 등에 기인하는 것으로 예상한다.

• Journal of Animal Science and Technology
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• 제62권2호
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• pp.227-238
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• 2020

Use of raw feedstuffs for livestock is limited by low digestibility. Recently, fermentation of feedstuffs has been highlighted as a new way to improve nutrient absorption through the production of organic acids using inoculated microorganisms, which can also play a probiotic role. However, standard procedures for feedstuff fermentation have not been clearly defined because the process is influenced by climatic variation, and an analytical standard for fermented feedstuffs is lacking. This study aimed to evaluate the microbiological and biochemical changes of feedstuffs during fermentation at temperatures corresponding to different seasons (10℃, 20℃, 30℃, and 40℃). We also investigated the effects of yeast, lactic acid bacteria (LAB), and Bacillus spp. on fermentation and determined the results of their interactions during fermentation. The viable cells were observed within 8 days in single-strain fermentation. However, when feedstuffs were inoculated with a culture of mixed strains, LAB were predominant at low temperatures (10℃ and 20℃), while Bacillus spp. was predominant at high temperatures (30℃ and 40℃). A significant drop in pH from 6.5 to 4.3 was observed when LAB was the dominant strain in the culture, which correlated with the concentrations of lactic acid. Slight ethanol production was detected above 20℃ regardless of the incubation temperature, suggesting active metabolism of yeast, despite this organism making up a marginal portion of the microbes in the mixed culture. These results suggested that fermentation temperature significantly affects microbiological profiles and biochemical parameters, such as pH and the lactic acid concentration, of fermented feedstuffs. Our data provide valuable information for the determination of industrial standards for fermented feedstuffs.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
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• pp.154-164
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• 2005

Saccharomyces cerevisiae KNU5377 is a thermotolerant strain, which can ferment ethanol from wasted papers and starch at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. This strain showed alcohol fermentation ability to convert wasted papers 200 g (w/v) to ethanol 8.4% (v/v) at 40$^{\circ}C$, meaning that 8.4% ethanol is acceptable enough to ferment in the industrial economy. As well, all kinds of starch that are using in the industry were converted into ethanol at 40$^{\circ}C$ with the almost same rate as at 30$^{\circ}C$. Hyperthermic cell killing kinetics and differential scanning calorimetry (DSC) revealed that exponentially growing cells of this yeast strain KNU5377 were more thermotolerant than those of S. cerevisiae ATCC24858 used as a control. This intrinsic thermotolernace did not result from the stability of entire cellular components but possibly from that of a particular target. Heat shock induced similar results in whole cell DSC profiles of both strains and the accumulation of trehalose in the cells of both strains, but the trehalose contents in the strain KNU5377 were 2.6 fold higher than that in the control strain. On the contrary to the trehalose level, the neutral trehalase activity in the KNU5377 cells was not changed after the heat shock. This result made a conclusion that though the trehalose may stabilize cellular components, the surplus of trehalose in KNU5377 strain was not essential for stabilization of whole cellular components. A constitutively thermotolerant yeast, S. cerevisiae KNU5377, was compared with a relatively thermosensitive control, S. cerevisiae ATCC24858, by assaying the fluidity and proton ATPase on the plasma membrane. Anisotropic values (r) of both strains were slightly increased by elevating the incubation temperatures from 25$^{\circ}C$ to 37$^{\circ}C$ when they were aerobically cultured for 12 hours in the YPD media, implying the membrane fluidity was decreased. While the temperature was elevated up to 40$^{\circ}C$, the fluidity was not changed in the KNU5377 cell, but rather increased in the control. This result implies that the plasma membrane of the KNU5377 cell can be characterized into the more stabilized state than control. Besides, heat shock decreased the fluidity in the control strain, but not in the KNU5377 strain. This means also there's a stabilization of the plasma membrane in the KNU5377 cell. Furthermore, the proton ATPase assay indicated the KNU5377 cell kept a relatively more stabilized glucose metabolism at high temperature than the control cell. Therefore, the results were concluded that the stabilization of plasma membrane and growth at high temperature for the KNU5377 cell. Genome wide transcription analysis showed that the heat shock responses were very complex and combinatory in the KNU5377 cell. Induced by the heat shock, a number of genes were related with the ubiquitin mediated proteolysis, metallothionein (prevent ROS production from copper), hsp27 (88-fold induced remarkably, preventing the protein aggregation and denaturation), oxidative stress response (to remove the hydrogen peroxide), and etc.