• Title/Summary/Keyword: yeast cells

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Studies on Takju Brewing with Potatoes (감자를 이용(利用)한 탁주제조(濁酒製造)에 관(關)한 연구(硏究))

  • Kim, S.Y.;Oh, M.J.;Kim, C.J.
    • Korean Journal of Agricultural Science
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    • v.1 no.1
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    • pp.67-81
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    • 1974
  • In order to prepare the mashing materials for "Takju", Korean wine, with potatoes they were steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows. 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D_{40^{\circ}}{^{30{\prime}}}$ 128 W.V. and 13.2 A. U. 3. The effects of various brewing conditions on the contents of Takju mashes wereas follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol fusel oil and Formal nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidities of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%) 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formal nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bateria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8$, $1.5{\times}10^8$) and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes (4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

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Activity and Survival of the Natural Bacteria under the Stressed Conditions Detected by Bioluminescent Phenotype (스트레스 하의 자연세균의 활성 및 생존의 발광표현형을 이용한 탐지)

  • Park, Kyoung-Je;Yoon, Hye-Young;Chun, Se-Jin;Lee, Ho-Sa;Lee, Dong-Hun;Jahng, Deokjin;Lee, Kyu-Ho
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.154-161
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    • 1998
  • To investigate whether the introduced genetic marker is useful to detect the survivalship and activity of the natural bacteria under the stressed conditions, one Gram-negative isolate, KP964 was transformed to the luminous phenotype by transferring luxAB gene. Under the starvation-stress this luminous bacterial culturability (determined by colony-forming-units [CFU] on agar plate) decreased rapidly below the detection limit by 37 days, while its total cell number (determined by AODC) remained almost the same as its initial inocular size. At that time period, the viable cell number was estimated to be 1400 times higher than its CFU number. The bioiuminescence (determined by relative light units [RLU]) produced under the same condition was also monitored and found to decrease more rapidly than the culturability by 5-fold. Under the other stresses, e.g., osmotic shocks, acid shock, and exposure to toxic chemicals, this bacterial strain did not show the reliable correlation between CFU and RLU. These results might not suggest the direct estimation of bioiuminescence from the stressed bacteria be an index of both the survivalship and its activity. However, when the stressed bacterial cells were incubated under the favorable condition by relieving from the existing stress, the potential bioiuminescence (the lag periods before the increase of bioiuminescence, the increase rates of bioiuminescence, and the maximal levels of bioiuminescence) was shown to be highly dependent upon the strengths of the stresses exposed to the bacterial cells. Therefore, analysis of the potential bioiuminescence from the stressed bacteria revealed good relationships with survival as well as activity.

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Physiological Changes of Saccharomyces cerevisiae KNU5377 Occurred in the Process of the 48-hour Ethanol Fermentation at 40℃ (40℃ 48시간 에탄올발효 과정 중 일어나는 Saccharomyces cerevisiae KNU5377의 생리 변화)

  • Kwak, Sun-Hye;Kim, Il-Sup;Kang, Kyung-Hee;Lee, Jung-Sook;Jin, Ingn-Yol
    • Journal of Life Science
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    • v.21 no.1
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    • pp.146-154
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    • 2011
  • In this study, physiological changes in a thermotolerant yeast Saccharomyces cerevisiae KNU5377 cell exposed to 48-hour alcohol fermentation at $40^{\circ}C$ were investigated. After 12 hours of alcohol fermentation at $40^{\circ}C$, the $C_{16:1}$ unsaturated acid of plasma membrane increased to 1.5 times more than the $C_{16:0}$ saturated fatty acid, and to about 2 times more for the $C_{18:1}$ unsaturated fatty acid. Fermentation at both $30^{\circ}C$ and $37^{\circ}C$ fermentation showed the same pattern as that done at $40^{\circ}C$. The pH of the alcohol-fermentation medium was reduced to pH 4.1 from a starting pH of 6.0 through the 12-hr fermentation and then maintained this level during the continuing fermentation. With the process of fermentation, the remaining glucose was reduced, but its amount remaining during the $40^{\circ}C$-fermentation was less reduced than those fermented at $30^{\circ}C$ and $37^{\circ}C$. In the study investigating the changing pattern of cellular proteins in the alcohol-fermenting cells, the SDS-PAGE and 2-D data indicated the most expressed dot was phosphoglycerate kinase, which is one enzyme involved in glycolysis. Why this enzyme was most expressed in the cells exposed to unfavorable conditions such as high temperature, increasing concentration of produced alcohol and long time exposure to other stress factors remains unsolved.

Brain-expressed X-linked 2 Binds to Kinesin Superfamily Protein 3A (Brain-expressed X-linked (Bex) 2와 heterotrimeric kinesin-2의 KIF3A와의 결합)

  • Kim, Mooseong;Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang Jin;Lee, Won Hee;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.32 no.3
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    • pp.189-195
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    • 2022
  • Kinesin-2 comprises two subfamilies of the heterotrimeric or homodimeric motors found in mammalian cells. Heterotrimeric kinesin-2 consists of kinesin superfamily proteins (KIFs) 3A and 3B and kinesin-associated protein 3 (KAP3), which is a molecular motor protein that moves along microtubules. It plays diverse roles in cargo transport, including anterograde trafficking in cilia, and interacts with many different cargoes and proteins, but their binding proteins have not yet been fully identified. In this study, the yeast two-hybrid assay was used to identify the proteins that interact with the cargo-binding domain (CBD) of KIF3A, and an interaction between KIF3A and brain expressed X-linked 2 (Bex2) was found. Bex2 bound to the CBD-containing C-terminal tail region of KIF3A but did not interact with the same region of KIF3B or KIF5A (a motor protein of kinesin-1). KIF3A interacted with another isoform, Bex1, but did not interact with Bex3. In addition, glutathione S-transferase (GST) pull-downs showed that KIF3A specifically interacts with GST-Bex1 and GST-Bex2 but not with GST alone. When co-expressed in HEK-293T cells, Bex2 co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. In combination, these results suggest that Bex2 is capable of binding to heterotrimeric kinesin-2 and may serve as an adaptor protein that links heterotrimeric kinesin-2 with cargo.

Changes in Microbial Counts, Enzyme Activity and Quality of Foxtail Millet Yakju Treated with High Hydrostatic Pressure During Storage (초고압 처리한 좁쌀약주의 저장 중 미생물수, 효소활성 및 품질변화)

  • 임상빈;좌미경;목철균;박영서
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.576-581
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    • 2004
  • Foxtail Millet Yakju were treated with heat ($65^{\circ}C$/15 min) or high hydrostatic pressure ($25^{\circ}C$ (RT) and $65^{\circ}C$ (HT)/400 MPa/10 min), and stored for 64 days at l$0^{\circ}C$, $25^{\circ}C$ and 37$^{\circ}C$. Changes in microbial counts, enzyme activity and quality of Yakju during storage were measured. Total viable cells were about 10$^2$ CFU/mL, and remained almost constant during storage at l$0^{\circ}C$ and $25^{\circ}C$, while decreased significantly at 37$^{\circ}C$, and undetected after 55 days of storage in heat- and pressure(RT)-treated, but after 25 days in pressure(HT)-treated Yakju. Lactic acid bacteria and yeast in heat- and pressure-treated Yakju were not detected during storage. The relative activities of a -amylase in heat- and pressure(RT)-treated were more than 100%, while those in pressure(HT)-treated were less than 40% during storage of 64 days at l$0^{\circ}C$. However, at $25^{\circ}C$ and 37$^{\circ}C$ the relative activities in untreated and pressure(RT)-treated were decreased greatly and then reached at the point of the activities of heat- and pressure(HT)-treated. The relative activities of glucoamylase in untreated and pressure(RT)-treated were decreased as the increase of the storage temperature during storage, while those in heat- and pressure(HT)-treated increased slightly as the increase of storage period at 1$0^{\circ}C$ and $25^{\circ}C$, and had no change at 37$^{\circ}C$. pH in heat- and pressure-treated had almost no change. Turbidity and reducing sugar in heat- and pressure-treated increased as the increase of storage temperature during storage.

Effect of Preparation method and Fermentation Conditions on Microbiological Characteristics of Sikhae (어류를 이용한 식해의 제조 방법에 따른 미생물의 특성변화)

  • Kim, Young-Sook;Oh, Seung-Hee;Kim, Soon-Dong
    • Food Science and Preservation
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    • v.15 no.6
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    • pp.909-914
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    • 2008
  • This study investigated microbial populations and the sensory quality of sikhae including globefish (GLS), flounder (FLS), gastropods (GAS) and whelks (WHS) during storage at 4C for 16 d following fermentation at 10C for 4 d. General bacterial numbers increased to 102 in GLS and FLS, and to 1045 in WHS and GAS after the 20 d fermentation/storage period. Lactic acid bacteria increased to 108 log cycle in GLS and FLS after 10 d ripening time, and reached this level in GAS and WHS after 15 d and 20 d, respectively. After 20 days the number of lactic acid bacteria in each of the four samples was 108. There were 104105 yeast cells/g in each of the four samples after 20 d. The number of Leuconostoc increased to over 108 log cycle after 10 d in GLS and FLS, and 15 days in GAS for WHS the increase was to 107 log cycle. The pH values of GLS, FLS, GAS and WHS 4.42, 4.56, 4.31 and pH 4.26, respectively. The Sikhae acidity for all four samples ranged from 1.551.85%. From the sensory evaluation the overall acceptability was in the order of FLS > GLS > GAS > WHS.

Quality Characteristics of Takju Fermentation by Addition of Chestnut Peel Powder (율피가루를 첨가한 탁주의 품질 특성)

  • Jeong Jin-Woong;Park Kee-Jai;Kim Myung-Ho;Kim Dong-Soo
    • Food Science and Preservation
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    • v.13 no.3
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    • pp.329-336
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    • 2006
  • The characteristics of mash qualities of takju prepared by addition of chestnut peel powder(5%, 10%, 20% and 30% per steamed rice) were investigated during fermentation. That is, in all fermentation periods, changes of pit total acid, organic acids, solids, amino nitrogen, total sugar and reducing sugar, microorganisms, alcohol and color were determined and analyzed. There was significant differences in characteristics of mash qualities by addition of chestnut peel powder. In general, contents of total acid, organic acids, amino nitrogen, total sugar, reducing sugar and ethanol of takju added with chestnut peel powder were lower than those of steamed rice only, whereas solid contents was higher. But ethanol content of takju added with 5% of chestnut peel powder after 8 days of fermentation was 9.6% which was similar to that of takju prepared by addition of steamed lice only. Also, microbial populations such as total viable cells, yeast and lactic acid bacteria of the treatments were increased to about $10^8CFU/mL$ after 2 days of fermentation and then decreased gradually. In the beginning stage of fermentation color differences value of the treatments were $1.99{\sim}10.27$, and the differentials reduced gradually during fermentation.

Establishment of Miniaturized Cultivation Method for Large and Rapid Screening of High-yielding Monascus Mutants, and Enhanced Production of Monacolin-K through Statistical Optimization of Production Medium (Monascus 균사체의 소규모 배양을 통한 고생산성 균주의 대규모 선별방법 확립과 통계적 생산배지 최적화를 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.305-312
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    • 2007
  • It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.

The physiological characteristics of Pleurotus ferulae Lanzi (아위버섯균의 생리적 특성)

  • Kim, Dae-Sik;Chai, Jung-Ki;Wi, Kye-Moon;Yoon, Dai-Ryoung;Oh, Deuk-Sil;Chang, Kyoung Soo;Son, Han-Gil
    • Journal of Mushroom
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    • v.2 no.2
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    • pp.88-96
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    • 2004
  • This study was executed to decide the physiological characteristics of Ferule mushroom. Four strains of Ferule mushroom were tested to select a superior strain in its mycelial growth. The pertinent substrates, temperature and pH ranges for the growth of selected strain were determined. And then, the wood rotting ability and type of the Ferule mushroom were determined. The superior strain F-2 among four strains was selected, on the basis of its vegetative mycelial growth and density on agar media. Mycelial growth of F-2 was the best on MYPA among other tested synthetic or semi-synthetic media. The temperature range for pertinent mycelial growth was about $25{\sim}34^{\circ}C$ and best at $30^{\circ}C$. The optimum pH range on MYPA was 5.0~6.0. The mycelial growth was mostly stimulated by soluble starch at cont. 1% (w/w) and secondly, maltose among several carabon sources and by mixed solution of YE(0.25%) and ME(0.25%) but not by ME alone. Cell thining and erosion of Pinus rigida wood by the mycelia of Ferule mushroom were found only on a few cell but largely at wood block test, indicating that the softwood rotting ability of Ferule mushroom mycelia was not so good. The result of polarized light microscopy appeared that cellulose of some tracheides showing the S3 layer lost brifringence was degraded by Ferule mushroom. But only part of cellulose of P. rigida wood was degraded by Ferule mushroom, because most of wood cells continued to showing briefingence. A largely degraded ray parenchyma and longitudinal parenchyma cell and partly thinning and erosion of hardwood(Quercus serrata) cell was found and it indicates that the rotting ability of Ferule mushroom mycelia on hardwood was higher than on softwood. It could be concluded that the difference in the wood rot by Ferule mushroom between the hardwood and softwood was made by the difference of chemical constitutions between them, especially in the contents and the types of lignin. Ferule mushroom was considered as white rotter as a result of bavendam test, although more research should be required.

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Cloning and Transcription Analysis of Sporulation Gene (spo5) in Schizosaccharomyces pombe (Schizosaccharomyces bombe 포자형성 유전자(spo5)의 Cloning 및 전사조절)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.112-118
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    • 2002
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.