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Production of ${\beta}-sitosterol$ by Cell Suspension Culture of Chrysanthemum coronarium L. (쑥갓세포의 현탁배양에 의한 ${\beta}-sitosterol$ 생산)

  • Kim, Hyun-Chul;Chung, Ha-Young;Lee, So-Youn;Chung, Ho-Yong;Kim, You-Jung;Baek, Nam-In;Kim, Soung-Hoon;Choi, Geun-Won;Kim, Dae-Keun;Kwon, Byoung-Mok;Park, Mi-Hyun;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.425-430
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    • 2005
  • [${\beta}-sitosterol$] is a plant sterol that reduces cholesterol levels and inhibits the growth of human prostate and colon cancer cells. Optimal conditions for ${\beta}-sitosterol$ production were examined from cell suspension cultures of Chrysanthemum coronarium L. The callus induction was optimal in MS medium containing 1 mg/l NAA and 1 mg/l BAP. Cell suspension culture was also established from the callus. Optimal ${\beta}-sitosterol$ production was obtained when the cells were cultured at an initial density of 2 mg DCW/l in MS medium containing 1 X sucrose (30 mg/l), 1 X nitrogen (1900 mg/l $KNO_3$, 1650 mg/l $NH_4NO_3$), and 1 X phosphate source (170 mg/l). In cell suspension cultures of C. coronarium L. using shake flasks, the peak content of ${\beta}-sitosterol$ was $150{\mu}g/g$ DCW. In cell suspension cultures of C. coronarium L. using an air-lift bioreactor, the maximum ${\beta}-sitosterol$ content of $143.8{\mu}g/g$ DCW was obtained at an air-flow rate of 100 cc/min.

Production Medium Optimization for Monascus Biomass Containing High Content of Monacolin-K by Using Soybean Flour Substrates (기능성 원료를 기질로 이용하는 Monacolin-K 고함유 모나스커스 균주의 생산배지 최적화)

  • Lee, Sun-Kyu;Chun, Gie-Taek;Jeong, Yong-Seob
    • KSBB Journal
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    • v.23 no.6
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    • pp.463-469
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    • 2008
  • During the last decade, monacolin-K biosynthesized by fermentation of red yeast rice (Monascus strains) was proved to have an efficient cholesterol lowering capability, leading to rapid increase in the market demand for the functional red yeast rice. In this study, the production medium composition and components were optimized on a shake flask scale for monacolin-K production by Monascus pilosus (KCCM 60160). The effect of three different soybean flours on the monacolin-K production were studied in order to replace the nitrogen sources of basic production medium (yeast extract, malt extract and beef extract). Among the several experiments, the production medium with dietary soybean flour to replace a half of yeast extract was very good for monacolin-K production. Plackett-Burman experimental design was used to determine the key factors which are critical to produce the biological products in the fermentation. According to the result of Plackett-Burman experimental design, a second order response surface design was applied using yeast extract, beef extract and $(NH_4)_2SO_4$ as factors. Applying this model, the optimum concentration of the three variables was obtained. The maximum monacolin-K production (369.6 mg/L) predicted by model agrees well with the experimental value (418 mg/L) obtained from the experimental verification at the optimal medium. The yield of monacolin-K was increased by 67% as compared to that obtained with basic production medium in shake flasks.

Studies on the Constituents and Culture of the Higher Fungi of Korea(II) -The Antitumor Components and Culture of Lentinus edodes(Berk.) Singer- (한국산(韓國産) 고등균류(高等菌類)의 성분(成分) 및 배양(培養)에 관한 연구(硏究)(II) -표고버섯의 항암성분(抗癌成分) 및 배양(培養)-)

  • Chung, Kyeong-Soo
    • The Korean Journal of Mycology
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    • v.10 no.1
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    • pp.33-39
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    • 1982
  • Carpophores of ten Korean strains of Lentinus edodes (Berk.) Singer, an antitumor polysaccharide producing fungus, were extracted with 0.1N NaOH solution. The extracts were dialized for seven days in distilled water and lyophilized to produce crude polysaccharide powders. Thus obtained crude polysaccharide samples were assayed for sugar contents by colorimetric method with anthrone reagent. Among ten strains examined Lentinus edododes-DMC7 was found to be the richest strain in polysaccharide content of carpophores. By shake culture experiment for biomass production, L. edodes-DMC7 was found to be the second most productive strain among seven strains examined. Cultural characteristics of L. edodes-DMC7 were investigated by shake culture method. The best result was obtained when L. edodes-DMC7 was cultured in the medium containing glucose 8g, starch 80g, yeast extract 12g, $KH_2PO_4\;0.87g,\;MgSO_4{\cdot}7H_2O\;O.5g,\;CaCl_2\;0.3g,\;FeSO_4{\cdot}7H_2O\;10mg\;ZnSO_4{\cdot}7H_2O\;4mg,\;CuSO_4{\cdot}5H_2O\;lmg,\;MnCl_2{\cdot}4H_2O\;7mg\;per\;11\;at\;28^{\circ}C$, 180 rpm, for 12 days. Thus thirty-three grams of dry mycelia was obtained per one liter of medium.

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Enhancement of Excretory Production of an Exoglucanase from Escherichia coli with Phage Shock Protein A (PspA) Overexpression

  • Wang, Y.Y.;Fu, Z.B.;Ng, K.L.;Lam, C.C.;Chan, A.K.N.;Sze, K.F.;Wong, W.K.R.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.637-645
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    • 2011
  • Production of recombinant proteins by excretory expression has many advantages over intracellular expression in Escherichia coli. Hyperexpression of a secretory exoglucanase, Exg, of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of E. coli. In this study, we demonstrated that overexpression of the PspA in the JM101(pM1VegGcexL-pspA) strain enhanced excretion of Exg to 1.65 U/ml using shake-flask cultivation, which was 80% higher than the highest yield previously obtained from the optimized JM101(pM1VegGcexL) strain. A much higher excreted Exg activity of 4.5 U/ml was further achieved with high cell density cultivation using rich media. Furthermore, we showed that the PspA overexpression strain enjoyed an elevated critical value (CV), which was defined as the largest quotient between the intracellular unprocessed precursor and its secreted mature counterpart that was still tolerable by the host cells prior to the onset of cell death, improving from the previously determined CV of 20/80 to the currently achieved CV of 45/55 for Exg. The results suggested that the PspA overexpression strain might tolerate a higher level of precursor Exg making use of the SecYEG pathway for secretion. The reduced lethal effect might be attributable to the overexpressed PspA, which was postulated to be able to reduce membrane depolarization and damage. Our findings introduce a novel strategy of the combined application of metabolic engineering and construct optimization to the attainment of the best possible E. coli producers for secretory/excretory production of recombinant proteins, using Exg as the model protein.

Study of Examples for Air Bag Non-deployment Including Rear Collision and Failure Phenomenon by Damage of Control Parts in Vehicle Air Bag (자동차 에어백의 제어부품 불량에 의한 고장현상 및 후방 추돌에 관련된 에어백 미전개에 대한 사례 연구)

  • Lee, Il Kwon;Kim, Young Gyu;Moon, Hak Hook
    • Journal of the Korean Institute of Gas
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    • v.16 no.6
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    • pp.102-106
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    • 2012
  • The purpose of this paper is to study the failure cases in relation to system of Air Bag in vehicle happened in the field. In the first example, it was separated the soldering parts connected the wire pin between air bag module and clock spring of air bag. Whenever the pin shake by the car's vibration, the driver verified the malfunction phenomenon appeared air bag warning lamp on instrument panel in front of driver's seat. in car inside room. The second example, it verified the warning lamp lighting phenomenon of air bag by produced the circuit plate non-contacting of single an element in air bag electronic control unit. The third example, it verified the light of air bag warning indicator lamp by separated with soldering parts connecting inner pin and resistance terminal of seat belt pretensioner using passenger seat. The fourth example, when the passenger car crash a back of truck, the former bumper get jammed under the latter as the roof height of car low less than that. Therefore, the impact of Car's collision verified that don't transfer with body frame of vehicle because of no attachment impact sensor in it.

고분자량의 pullulan 생산을 위한 발효공정의 최적화

  • Kim, Seong-Gu;Lee, Ji-Hyeon;Kim, Jeong-Hwa;Kim, Mi-Ryeong;Lee, Jin-U
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.45-50
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    • 2000
  • For the maximum production of pullulan from glucose as a carbon source, the effects of glucose concentration, pH and dissolved oxygen concentration on the cell growth and mass production of high-molecular weight pullulan by A. pullulans ATCC 42023 were evaluated. A. pullulans showed optimum pullulan productivity when glucose concentration was 0.3M (54g/L). And inhibitory effects on the cell growth and the pullulan production were observed at the glucose concentration higher than 0.3M (54g/L). The influence of pH control and dissolved oxygen on the pullulan production and growth of A. pullulans was studied. In shake-flasks, maximum pullulan production was obtained with $11.98g/{\ell}$ when initial pH was 6.5. In the batch fermentation, the maximum pullulan production of $13.31g/{\ell}$ was obtained with constant pH 4.5. And it was found that pullulan yield and synthesis rate increased with oxygen availability. For the production of commercially useful pullulan with high-molecular weight, a mixed carbon source, which was a mixture of glucose and glucosamine, was used for the pullulan fermentation with A. pullulans. On the basis of 5% mixed carbon source, culture with 3% glucosamine with 2% glucose was optimum condition for the production of high (M.W.> 1,000,000) and medium (M.W.> 200,000) molecular weight pullulan with considerable yields of cell mass and product. And the influence of pH control on the molecular weight of pullulan was studied in batch fermentation. It was found that the productivity of high-molecular weight pullulan with pH control at 6.5 was higher than that with no pH control.

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Favorable Conditions for Mycelial Growth of Phellinus linteus (목질진흙버섯(Phellinus linteus)의 적합한 균사생장)

  • Lee, Won-Ho;Kim, Su-Young;Park, Young-Jin;Kim, Tae-Woong;Kim, Ho-Kyung;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.95-100
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    • 2004
  • The main objectives of the study were to investigate cultural characteristics of Phellinus linteus. The optimum culture media for mycelial growth of P. linteus were MYA (malt yeast agar) and SMS (soybean powder malt Sucrose). Similarly, optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. Malt extract (2%, v/v) and yeast extract (0.2%, v/v) were optimum carbon and nitrogen sources. Similarly, 0.1% $KH_2PO_4$ was optimum mineral salt. Highest mycelial growth was observed when C/N ratio was 10 : 1. Optimum inoculum amount for flask culture was $5{\sim}6$ mycelial discs (6 mm diameter) per 100 ml of liquid medium, Highest mycelial dry weight was obtained when cultured in 100 ml liquid medium in 300 ml shaking flask after 20 days of shaking culture, For mass liquid culture (8 l), flask culture was homogenized and used as an inoculum. Optimum culture period and aeration rate for 8l fermentation culture were 12 days and 2.0 vvm, respectively.

Studies on the chlorella in Korea (한국 Chlorella에 대한 연구)

  • 이주식;심종호
    • Korean Journal of Microbiology
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    • v.1 no.1
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    • pp.38-44
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    • 1963
  • According to the experiment on pure-isolation and the related contaminants of Chlorella, the phenomena of the ecological distributions of Chlorella in Korea have been manifested in several areas and also the aim that in going to do culture, biological and physiological study of Chlorella is carried out. Contaminants very oftenly occupied on the colony of the strains taken in order to fulfil pure-isolation of Chlorella, but in accordance with being piled up the minute research on this subject, I can obtain the desirable results as follows: 1. For the pure-isolation, the duration chose the time from May to September 1957 so that may easily isolate from contaminant water with utilizing the antibiotic substances. 2. To take long time, 36-48 hours until growth of nascent through the non-sporulated, it originates from the difference of the cultured media. In addition to the above mention, the mechanism of growth until nascent through the sporulated must not always require the ligh. However the supply of metabolic energy depend upon its nutritional conditions per phase. 3. The culture of Chlorella should be based on the lower culturing except adding especial conditions such as reagent concentration of media, artifical shake of media and other facts due to the natural conditions. And also these strains grew not only in distilled water but 2% NaCl solution without any abnormality in cell it self. I, therefore, guess it is possible to culture in sea-water under phasic environment. 4. In the experiment of ammonia detection, it is caused by the sampling surroundings to contain the minute quantity of ammonia in strain No. M 918; that is the place to be plenty of Carbohydrate on behalf of protein. 5. To compare the absorption curve of chlorophyll of higher plant with that of Chlorella, the absorption zone made mostly the Same ones each other but a little absorption grade dose not clearly appear. The colony which formed giant type grows with intensive colour and green band on surrounding of the colony and after that it was changed into all the green colour and developed up to end. 6. At first phase for a week, the development of Chlorella suspends the normal condition as in vivo but after a few days, the colour of chlorophyll gradually changed into blue-yellow which secrete the mucous substances on the agar media. The cell was flew out the contained substances itself on leaving the cell wall only, or the various micro-organism diffused on the outer-region of the cell.

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Influence on Lysine Production by Overexpression of the ddh Gene in a Lysine-producing Brevibacterium lactofermentum (Brevibacterium lactofermentum에서 ddh 유전자의 Overexpression이 $_L-Lysine$ 생산에 미치는 영향)

  • Park, Sun-Hee;Kim, Ok-Mi;Kim, Hyun-Jeong;Bae, Jun-Tae;Chang, Jong-Sun;Lee, Kap-Rang
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.224-230
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    • 1999
  • The ddh gene encoding meso-DAP-dehydrogenase (DDH) involved in the dehydrogenase pathway is essential for high-level lysine production in Brevibacterium lactofermentum. To investigate its influence on lysine production by overexpression of the ddh gene in a lysine-producing B. lactofermentum, recombinant plasmid pRK1 and pRK31 containing the ddh gene of B. lactofermentum were constructed and they were introduced into B. lactofermentum by electroporation. Multiple copies of pRK1 and pRK31 caused 7-fold and 14-fold increase of DDH activity in B. lactofermentum cell extracts, respectively. As determined in shake flask fermentation, lysine production of B. lactofermentum harboring pRK1 or pRK31 was 22% or 19% higher than that of the control, respectively.

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Verifying ASCE 41 the evaluation model via field tests of masonry infilled RC frames with openings

  • Huang, Chun-Ting;Chiou, Tsung-Chih;Chung, Lap-Loi;Hwang, Shyh-Jiann;Jaung, Wen-Ching
    • Earthquakes and Structures
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    • v.19 no.3
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    • pp.157-174
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    • 2020
  • The in-situ pushover test differs from the shake-table test because it is performed outdoors and thus its size is not restricted by space, which allows us to test a full-size building. However, to build a new full-size building for the test is not economical, consequently scholars around the world usually make scale structures or full-scale component units to be tested in the laboratory. However, if in-situ pushover tests can be performed on full-size structures, then the seismic behaviors of buildings during earthquakes can be grasped. In view of this, this study conducts two in-situ pushover tests of reinforced concrete (RC) buildings. One is a masonry-infilled RC building with openings (the openings ratio of masonry infill wall is between 24% and 51%) and the other is an RC building without masonry infill. These two in-situ pushover tests adopt obsolescent RC buildings, which will be demolished, to conduct experiment and successfully obtain seismic capacity curves of the buildings. The test results are available for the development or verification of a seismic evaluation model. This paper uses ASCE 41-17 as the main evaluation model and is accompanied by a simplified pushover analysis, which can predict the seismic capacity curves of low-rise buildings in Taiwan. The predicted maximum base shear values for masonry-infilled RC buildings with openings and for RC buildings without masonry infill are, respectively, 69.69% and 87.33% of the test values. The predicted initial stiffness values are 41.04% and 100.49% of the test values, respectively. It can be seen that the ASCE 41-17 evaluation model is reasonable for the RC building without masonry infill walls. In contrast, the analysis result for the masonry infilled RC building with openings is more conservative than the test value because the ASCE 41-17 evaluation model is limited to masonry infill walls with an openings ratio not exceeding 40%. This study suggests using ASCE 41-17's unreinforced masonry wall evaluation model to simulate a masonry infill wall with an openings ratio greater than 40%. After correction, the predicted maximum base shear values of the masonry infilled RC building with openings is 82.60% of the test values and the predicted initial stiffness value is 67.13% of the test value. Therefore, the proposed method in this study can predict the seismic behavior of a masonry infilled RC frame with large openings.