• Title/Summary/Keyword: Stationary phase

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Simultaneous HPLC Determination of Marker Compounds for the Standardization of Hedyotis diffusa (백운풀의 지표성분 설정 및 품질표준화를 위한 정량 분석법)

  • Bang, Han-Yeol;Yang, Eun-Ju;Kim, Jeong-Ah;Song, Kyung-Sik
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1025-1031
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    • 2013
  • From a 95% ethanolic extract of H. diffusa, four marker compounds (HD1~HD4) were isolated, which were relatively unique and exist in comparably high contents. The structures of marker compounds were identified as digitolutein (1), 2-hydroxy-3-methylanthraquinone (2), (E/Z)-6-O-p-coumaroyl scandoside methyl ester (4:1 mixture) (3), and (E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester (4:1 mixture) (4), respectively, on the basis of $^{13}C$ and $^1H$-NMR analyses. The calibration curves of marker compounds showed high linearity, as their correlation coefficient ($R^2$) were in the range of 0.9991~0.9999. In addition, the limit of detection (LOD) and the limit of quantification (LOQ) were $0.03{\sim}0.07{\mu}g/ml$ and $0.099{\sim}0.231{\mu}g/ml$, respectively. The intra-day/inter-day precision and accuracy were 0.23~2.00%/0.25~1.16% and 94.60~108.44%/94.73-110.23%, respectively. The optimal HPLC conditions for the simultaneous quantification of HD1~HD4 were as follows: stationary phase; Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), column temp.; room temperature, UV detection at 280 nm, flow rate; 2.0 ml/min, injection volume; $10{\mu}l$, mobile phase; start with the mixture of 80% solvent A ($H_2O$ containing 0.5% acetic acid) and 20% solvent B (methanol containing 0.5% acetic acid) and gradually decrease solvent A to 40% in 9 min., then retain this condition to 18 min. Under the HPLC condition, the four marker compounds 1~4 were successfully separated without any interference of other constituents. The results obtained in this study are expected to be helpful for the development of nutraceutics and natural medicines and for the quality control of this plant.

Studies on the Brewing of Apple Wine -Culture Conditions of a Cider Yeast, Saccharomyces sp. R-11 on the Synthetic Medium (사과주(酒) 양조(釀造)에 관한 연구(硏究) -사과주효모(酒酵母) Saccharomyces sp. R-11의 합성배지((合成培地)에서의 배양(培養) 조건(條件)-)

  • Chung, Ki-Taek;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.10 no.2
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    • pp.75-83
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    • 1982
  • As a primary study for cell growth and alcohol production of a cider yeast, Saccharomyces sp. R-11, cultural and nutritional characteristics of the strain were investigated. The results obtained were as follows: The optimum culture medium for this strain was a synthetic medium, Henneberg B, and sucrose was the best carbon source for yeast growth and alcohol production. Optimum sugar concentrations for yeast growth and alcohol production were 15% and 25%, respectively. Optimum pH and temperature of the basal medium for growth of this strain were 4.5 and $30^{\circ}C$ respectively. The yeast growth was enhanced by the addition of 100 ppm of $Mg^{2+}$, but significantly inhibited by the addition of 100 ppm of $Co^{2+}$. Lower temperature and maintenance of optimum pH for yeast growth increased the final alcohol concentration. Under optimum condition for cell growth at stationary culture, generation time and specific growth rate of the strain were 7.5 hr and 0.092 $hr^{-1}$, respectively. At 8% initial alcohol concentration, yeast growth was inhibited about 50% and this strain could not be grown at more than 12% initial alcohol. The strain could be grown at less than 125ppm $SO_2$without alcohol addition, and at less than 75 ppm $SO_2$ with 8% initial alcohol. The higher sulfur dioxide concentration of a medium, the longer lag phase in yeast growth was observed. This strain could induced alcoholic fermentation at less than 10% initial alcohol concentration with 0 and 25 ppm $SO_2$, at less than 8% initial alcohol with 50 and 75 ppm $SO_2$, and at less than 6% initial alcohol with 100 and 125 ppm $SO_2$.

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Effect of Water Uptake Rate on Germination Characteristics of Waxy Rice Seeds and Guaiacol Peroxidase Activity during Early Imbibition (종자의 수분흡수속도가 찰벼 품종별 발아특성과 침윤초기 Guaiacol Peroxidase활성에 미치는 영향)

  • Shon, Jiyoung;Kim, Junhwan;Lee, Chung-Kuen;Yang, Woonho;Yoon, Young-Hwan;Chung, Nam-Jin;Kim, Chung-Kon;Kim, Bo-Kyung;Choi, Kyung-Jin
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.4
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    • pp.416-423
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    • 2013
  • Germination is controlled by internal factors of seeds and external factors such as water, temperature and light. We investigated the relationship between germination characteristics of four waxy rice cultivars and patterns of water uptake, antioxidant enzymes and free soluble sugars during early imbibition. Seed viabilities by tetrazolium test of four different rice cultivars were higher than 95% and germination rates of the hulled rice seeds were on 95% average. However, germination rate of intact rice among four cultivars showed a big difference depending on temperature. Water uptake of hulled and intact rice seeds during imbibition reached a stationary phase at around 30% moisture content. Although rates of water uptake were faster in hulled rice and high temperature than intact rice and low temperature condition, difference of those among cultivars was greater under low temperature than high temperature. The time required for rice seeds to uptake 30% water was negatively correlated with percentage of germination, germination energy, germination speed and mean germination time. Guaiacol peroxidase activity at 24h of imbibition was correlated with germination energy and germination speed but not percentage of germination. Catalase activity, soluble protein and maltose concentration at 24h of imbibition were not correlated with characteristics of germination. These results suggest that a time required for rice seeds to uptake 30% of water significantly correlated with germination and guaiacol peroxidase activity during early imbibition plays an important role in initiation of germination.

Effect of Oregano (Origanum vulgare L.) on the Survival of Escherichia coli ) O157:H7 and Staphylococcus aureus 196E during Cold Storage (저온저장중 Oregano(Origanum vulgare L.)가 Eschetichia coli O157:H7과 Staphylococcus aureus 196E의 생존에 미치는 영향)

  • 박찬성;박금순;김미림
    • Korean journal of food and cookery science
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    • v.13 no.4
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    • pp.440-447
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    • 1997
  • The antibacterial effect of low concentrations of oregano (Origanum vulgare L.) in culture broth against Escherichia coli O157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of oregano was inoculated with 10$\^$6/∼10$\^$7/ CFU/$m\ell$ of E. coli or S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% oregano and the growth occured at 2% oregano but only after a prolonged lag period. The death rate of E. coli after stationary phase was increased with increasing concentration of oregano in culture broth. The growth of S. aureus was inhibited with increasing concentration of oregano at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.3∼0.5% oregano after a long lag period while the viability at 1.0∼2.0% was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli or S. aureus was increased with the progress of time and increasing spice concentration. At the presence of 0.5∼2.0% oregano, E. coli and S. aureus were dead after 20 and 16 days of refrigerated storage, respectively. During frozen storage at -20$^{\circ}C$, the antibacterial activity of oregano against E. coli was increased with increasing storage time and spice concentration while the antibacterial activity against S. aureus was effective during the early period of storage, and no changes in the population of S. aureus occurred at different concentrations of oregano during frozen storage. Viable counts of E. coli were 1/3∼l/7 and S. aureus were 1/18∼l/46 of the control at 0.1% oregano in culture broth during frozen storage.

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A Study on Mobile Antenna System Design with Tri-band Operation for Broadband Satellite Communications and DBS Reception (광대역 위성 통신/방송용 삼중 대역 이동형 안테나 시스템 설계에 관한 연구)

  • Eom Soon-Young;Jung Young-Bae;Son Seong-Ho;Yun Jae-Seung;Jeon Soon-Ick
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.17 no.5 s.108
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    • pp.461-475
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    • 2006
  • In this paper, it is described about the tri-band mobile antenna system design to provide broadband multimedia and direct broadcasting services using goo-stationary Koreasat 3, simultaneously operated in Ka/K/Ku band. The radiating part of the antenna system with a fan beam characteristic in the elevation plane is composed of the quasi-offset dual shaped reflector and the tri-band feeder. The tri-band feeder is also composed of the Ka/K dual band feeder with the protruding dielectric rod, the circular polarizer, the ortho-mode transducer and the circular-polarized Ku band feed array. Especially, the Ka/K dual band circular polarizer was realized firstly using the comb-type structure. For fast satellite-tracking on the movement, the Ku band feed array has the structure of the $2{\times}2$ active phased array which can make electrical beams. And, the circular-polarized characteristic in the feed array was improved by $90^{\circ}$ rotating arrangement of four radiating elements polarized circularly by a $90^{\circ}$ hybrid coupler, respectively. Four beam forming channels to make electrical beams at Ku band are divided into the main beam channel and the tracking beam channel in the output, and noise temperature characteristics of each channel were analyzed on the basis of the contributions of internal sub_units. From the fabricated antenna system, the output power at $P_{1dBc}$ of Ka_Tx channel was measured more than 34.1 dBm and the measured noise figures of K/Ku_Rx channels were less than 2.4 dB and 1.5 dB, respectively, over the operating band. The radiation patterns with co- and cross-polarization in the tri-band were measured using a near-field measurement in the anechoic chamber. Especially, Ku radiation patterns were measured after correcting each initial phase of active channels with partial radiation patterns obtained from the independent excitation of each channel. The antenna gains measured in Ka/K/Ku band of the antenna system were more than 39.6 dBi, 37.5 dBi, 29.6 dBi, respectively. And, the antenna system showed good system performances such as Ka_Tx EIRP more than 43.7 dBW and K/Ku_Rx G/T more than 13.2 dB/K and 7.12 dB/K, respectively.

Phytochrome Signal Transduction Regulates Anthocyanin Biosynthesis in Cell Suspension Cultures of Vitis vinifera (포도 세포현탁배양계에서 Phytochrome 신호전달에 의해 조절되는 안토시아닌 생합성)

  • Choi, Kwan-Sam;Kim, Sun-Kyung;In, Jun-Gyo;Shin, Dong-Ho
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.239-248
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    • 2004
  • This experiment was carried out to confirm that phytochrome regulates anthocyanin bio-synthesis during cell suspension culture system of grape or not. In suspension culture of grape, maximum accumulation of anthocyanin was observed at the stationary phase under continuous white light condition. From mono-chromatic light interruption for 24h at the 4th or 7th day on the suspension cultured cells, the anthocyanin accumulation was highly enhanced at the light interruption at 7th day than 4th day under all monochromatic light treatment. However, the cell growth patterns were not affected by any light treatment. In the darkness, the anthocyanin synthesis was very low but remarkably increased by blue light or red light irradiation. However, the increase of anthocyanin accumulation by blue or red light was suppressed by far-red light in the suspension cells of grape. This suppression by far-red light on the anthocyanin synthesis also observed on the cells treated red or far-red light alternatively. These results implied that phytochrome regulation system may be involved in the anthocyanin biosynthesis of the suspension grape cells. By RNA expression analysis, chalcone synthase (CHS) gene was expressed highly by blue and red light but low by far-red light. The synergistic increase of CHS gene expression was also observed at the treatment of blue light followed by red for 24h. This result may explain the increase of anthocyanin accumulation in B/R treatment. Although the expression of phytochrome gene (PHYA or PHYB) was not highly increased by all light treatment (blue, red, and far-red light) the expression of both PHYA gene and PHYB gene was increased a little in cells treated red or far-red light. In grape suspension cells, the red light enhanced the anthocyanin synthesis, whereas the far-red light was suppressed. Although it was not confirmed whether or not phytochrome gene is activated in anthocyanin accumulating grape cells, we believed that anthocyanin biosynthesis in grape cells may be regulated under phytochrome signal transduction system.

Effects of Aeration on Biological Activities During Composting of Dairy Manure in Enclosed BenchScale Reactor (밀폐형 Bench-scale reactor 에서의 우분 퇴비화시 Aeration 이 생물학적 활성에 미치는 영향)

  • Kang, Hang-Won;Zhang, R.H.;Park, Hyang-Mee;Ko, Jee-Yeon;Rhee, In-Koo;Park, Kyeong-Bae
    • Korean Journal of Environmental Agriculture
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    • v.17 no.3
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    • pp.260-267
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    • 1998
  • This experiment used the enclosed bench-scale reactors of 242 liters was conducted to obtain basic data on temporal and spatial variations in temperature, oxygen and moisture content, which were important factors of biological activities, during composting of mixture of dairy manure and rice straw. The reactors with thermocouples, oxygen sensor and datalogger were aerated at four different rates of 0.09, 0.18, 0.90 and 1.79 l $min^{-1}kg$ dry $solids^{-1}$. The higher aeration rates were, the faster the rates of increase and decrease in composting temperature were in both of initial and turnover stage, and the smaller the temperature difference between exhaust air and composting materials. Composting temperature of initial stage increased suddenly in all aeration rates, then stationary phase of temperature in materials and exhaust air showed at $50{\sim}53^{\circ}C$ for 5 hours and at $45^{\circ}C$ between 5 and 15 hours, respectively. In initial stage the maximum temperature was decreased with increasing aeration rates but in the stage after turnover it was the opposite except for 1.79 l $min^{-1}kg^{-1}$. Time arrived at the maximum temperature of composting materials was later in low-aeration rates than high-aeration rates at both stages. Time maintained high-temperature more than $45^{\circ}C$ was rapidly decreased with increasing aeration rates. In initial stage of composting maintaining time of $65^{\circ}C$ or more was the longest in the treatments of 0.09 and 0.18 l $min^{-1}kg{-1}$, while those of $55{\sim}65^{\circ}C$ and $45{\sim}55^{\circ}C$ was in 0.90 and 1.79 l $min^{-1}kg{-1}$, respectively. The minimum oxygen content and the maximum oxygen consumption rate in exhaust air through composting materials showed the increased trends with increasing aeration rates. In initial stage the minimum oxygen content was ranged from 0.9% to 7.4% for 32 to 59.5 hours and the maximum oxygen consumption rate was $1.89{\sim}6.48$ $gh^{-1}kgVS^{-1}$. In the stage after turnover their levels were $2.1{\sim}19.9%$ and $1.76{\sim}3.49 %$g/h-㎏ VS, respectively, for 16 to 49.5 hours.

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Studies on the Production of Intra- and Extra-cellular Lipids by the Strains in the Genus RHODOTORULA (Rhodotorula 속(屬) 균주(菌株)에 의(依)한 세포(細胞) 내외(內外) 지질생산(脂質生産)에 관(關)한 연구(硏究))

  • Park, Sung-Oh
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.93-116
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    • 1974
  • A potent intracellular-lipid-producing yeast, Rhodotorula glutinis var. glutinis SW-17, was screened out from a variety of arable soils, compost heaps, and fodders, and two strains of excellent extracellular-lipid-producing yeasts, Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54, were screened out from the surface of many species of leaves. And then the intra- and extra-cellular lipid productions by those Rhodotorula yeasts were studied. The results were as follows: 1. During the shaking culture of 8 days at $24^{\circ}C$, both the intra- and extra-cellular lipid accumulation started almost at the stationary phase of growth, when the nitrogen source in the medium was a little more than half used up. The intracellular lipid production by Rhodotorula glutinis var. glutinis SW-17 reached 58.42% (w/w) of dried yeast, and the extracellular lipid production by Rhodotorula graminis SW-54 amounted to 2.62g per liter of the medium. 2. After the carbon and nitrogen sources in the medium were almost consumed, if the yeasts were shake-cultured further in a state of starvation, the yeast cells re-utilized the already produced intra- and extra-cellular lipids and the lipids completely disappeared in the medium in about 90 days. 3. The relative concentration of carbon and nitrogen sources in the media greatly influenced both the intra- and extra-cellular lipid production. When the nitrogen source in the medium was almost used up for the growth of yeast, and excess carbon sources were still available, the lipid production vigorously proceeded. As long as the nitrogen source concentration in the medium was high, the lipid production was greatly suppressed. 4. The optimum pH for both the intra- and extra-cellular lipid production by those yeasts was pH 5.0-6.0. 5. The fatty acid components of the intracellular lipid of Rhodotorula glutinis var. glutinis SW-17 were myristic, palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic acids. The largest components of the fatty acids were palmitic acid equivalent to 30-45% of the whole fatty acids and oleic acid equivalent to 35-50%. 6. The fatty acid components of the extracellular lipid of Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54 were myristic, palmitic, stearic, oleic, linoleic, linolenic, 3-D-hydroxypalmitic, and 3-D-hydroxystearic acids. The largest components of the fatty acids were 3-D-hydroxypalmitic acid equivalent to 22-25% of the acids and 3-D-hydroxystearic acid equivalent to 13-17%. 7. The polyol component of the intracellular lipids was only glycerol, whereas the polyols of extracellular lipids were glycerol, mannitol, xylitol and arabitol.

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Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Development of Saccharomyces cerevisiae Strains with High RNA Content (리보핵산을 다량으로 함유하는 Saccharomyces cerevisiae 균주의 개발)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Min, Byoung-Cheol;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.465-474
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    • 1999
  • RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

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