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Enhanced Production of Oleanolic Acid by the Elicitation in Oldenlandia diffusa Suspension Cell Cultures (백화사설초의 현탁세포배양에서 Elicitation에 의한 Oleanolic acid 생산성 증대)

  • Lee Yong-Il;Kim Dong-Il
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.471-477
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    • 2004
  • Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

Free Radical Scavenging Effect and Extraction Condition of Ethanol Extracts and Fractions of Wild Grape Seed (Vitis coignetiea) (머루종자의 적정 추출조건 및 추출 분획물의 Free Radical 소거능 효과)

  • Kim, Nan-Young;Kim, Young-Kuk;Bae, Ki-Ja;Choi, Jae-Ho;Moon, Jea-Hak;Park, Geun-Hyung;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.755-758
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    • 2005
  • Wild grape is a traditional medicine plant in north-eastern part of Asia and has been known to have healing properties for various illnesses. This study was to determine the optimum extraction condition and antioxidant activity of ethanol extracts of wild grape (V. coignetiae) seed. Also, organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of wild grape seed at different temperatures. Total ethanol extraction yield of wild grape seed ranged from $4\%\;to\;12\%$ depending on the ethanol concentration, extraction temperature and time condition. The highest extraction yield of $11.9\%$ was obtained at $90\%$ ethanol condition for 12 hour at $70^{\circ}C$. However, the strongest free radical scavenging effect $(RC_{50})$ with $20.93\mu g/mL$ was observed in $70\%$ ethanol extract of wild grape seed extracted for 6 hour at $70^{\circ}C,\;while\;RC_{50}\;with\;40.42$\mu g/mL$ was observed in $90\%$ ethanol extract for 12 hour at $70^{\circ}C$. Antioxidant activity of ethanol extracts of wild grape seed increased as total phenol contents increased. Among each fraction obtained from organic solvents, ethyl acetate fraction was found to have the strongest $RC_{50}\;(8.6\mu g/mL)$ and 636.77 mg GAE/g phenol contents .

Enhanced production of monacolin-K through supplement of monacolin-K precursors into production medium and cloning of SAM synthetase gene (metK) (Precursor제공 및 생합성 관련 유전자의 cloning을 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.23 no.6
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    • pp.519-524
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    • 2008
  • Monacolin-K is a strong anti-hypercholesterolemic agent produced by Monascus sp. via polyketide pathway. High-yielding mutants of monacolin-K were developed through rational screening strategies adopted based on understanding of monacolin-K biosynthetic pathway. Through the experiments for investigating various amino acids as putative precursors for the monacolin-K biosynthesis, it was found that production level of monacolin-K was remarkably increased when optimum amount of cysteine was supplemented into the production medium. We suggested that these phenomena might be related to the special roles of SAM (S-adenosyl methionine), a putative methyl group donor in the biosynthetic pathway of monacolin-K, demonstrating close interrelationship between SAM-synthesizing primary metabolism and monacolin-K synthesizing secondary metabolism. Namely, increase in the intracellular amount of SAM derived from the putative precursor, cysteine which was extracellularly supplemented into the production medium might contribute to the significant enhancement in the monacolin-K biosynthetic capability of the highly mutated producers. On the basis of these assumptions derived from the above fermentation results, we decided to construct efficient expression vectors harboring SAM synthetase gene (metK) cloned from A. nidulans, with the hope that increased intracellular level of SAM could lead to further enhancement in the monacolin-K production through overcoming a rate-limiting step associated with monacolin-K biosynthesis. Hence, in order to overcome the plausible rate-limiting step associated with monacolin-K biosynthesis by increasing intracellular level of SAM, we transformed the producer mutants with an efficient expression vector harboring gpdA promoter of the producer microorganism, and metK gene. Notably, from the resulting various transformants, we were able to screen a very high-yielding transformant which showed approximately 3.3 fold higher monacolin-K productivity than the parallel nontransformed mutants in shake flask cultures performed under the identical fermentation conditions.

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.

Treatment of Malodorous Waste Air by a Biofilter Process Equipped with a Humidifier Composed of Fluidized Aerobic and Anoxic Reactor (폐가스 가습조(유동상호기 및 무산소조)를 포함한 바이오필터공정을 이용한 악취폐가스의 처리)

  • Lim, Kwang-Hee
    • Korean Chemical Engineering Research
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    • v.56 no.1
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    • pp.85-95
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    • 2018
  • In this research, a biofilter system equipped with a biofilter process and a humidifier composed of a fluidized aerobic and an anoxic reactor, was constructed to treat odorous waste air containing hydrogen sulfide, ammonia and VOC, frequently generated from pig and poultry housing facilities, compost manufacturing factories and publicly owned facilities. Its optimum operating condition was revealed and discussed. In the experiment of complex feed, the ammonia of fed-waste air was removed by ca. 75% and more than 20% at the stage of the humidifier and the biofilter, respectively. The toluene of the fed-waste air was removed by ca. 20% and more than 70% at the stage of the humidifier and the biofilter, respectively. Therefore the water-soluble ammonia and the water-insoluble toluene were treated mainly at the stage of the humidifier and the biofilter, respectively. In addition, hydrogen sulfide was almost absorbed at the stage of the humidifier so that it was not detected at the biofilter process. In the experiment of ammonia-containing feed, the ammonia of fed-waste air was removed by ca. 65% and 35% at the stage of the humidifier and the biofilter, respectively. Its removal efficiency of ammonia at the stage of the humidifier was 10% less than that in the experiment of complex feed, due to no supply of such carbon source as toluene required in the process of denitrification. In the experiments of complex feed, ammonia-containing feed with and without (instead, glucose) the addition of yeast extract, the absorption rates of ammonia-nitrogen were ca. 0.28 mg/min, 0.23 mg/min and 0.27 mg/min, respectively. The corresponding denitrification rates in the anoxic reactor were 0.42 mg/min, 0.55 mg/min and 0.27 mg/min, respectively. In addition, in the modeling of bubble column(the fluidized aerobic reactor of the humidifier) process, the value of specific surface area(a) of bubbles multiplied by enhanced mass transfer coefficient (E $K_y$) was evaluated to be 0.12/hr.

Control of Irrigation Amount for Production of High Quality Fruit in Melon Fertigation Cultivation (멜론 관비재배시 고품질 과실생산을 위한 관수량 조절)

  • Rhee, Han-Cheol;Cho, Myeung-Whan;Eom, Young-Cheol;Park, Jin-Meun;Lee, Jae-Han
    • Journal of Bio-Environment Control
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    • v.17 no.4
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    • pp.288-292
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    • 2008
  • This study was conducted to identify the effects of irrigation amount to produce high quality melon fruit in fertigation culture. Irrigation amount of during fruit harvesting period was doubled at the low irrigation point ($(-45{\sim}50\;kPa$) treatment as 115 mm as than that of the high irrigation point ($-20{\sim}25\;kPa$) treatment. The plant growth rates such as stem length, leaf weight and plant height were a little diminished at the low irrigation point ($-45{\sim}50\;kPa$) than those of the other treatments. Internode length was however not affected by irrigation amount. Fruit weight was lighter at the low irrigation point ($-45{\sim}50\;kPa$) than that of at the high irrigation point and fruit height was shorter, but fruit diameter was not affected by irrigation amount. Fruit soluble solid was $0.9^{\circ}Bx$ higher at the low irrigation point ($-45{\sim}50\;kPa$) than at the high irrigation point ($-20{\sim}25\;kPa$) and net index was higher. Total marketable yield was highest by 3,937 kg/10a at the high irrigation point ($-20{\sim}25\;kPa$), but the excellent marketable yield was highest by 2,531 kg/10a at the low irrigation point ($-45{\sim}50\;kPa$). Inorganic contents of the soil N, K, Ca and Mg were not affected by irrigation amount. It was therefore thought that optimum irrigation point to produce high quality melon fruit by fertigation culture was $-45{\sim}50\;kPa$ at ripening stage.

Growth Response of Lettuce to Various Levels of EC and Light Intensity in Plant Factory (배양액 농도와 광도가 식물공장에서 재배되는 적축면 상추의 생장에 미치는 영향)

  • Cha, Mi Kyung;Kim, Ju-Sung;Cho, Young Yeol
    • Journal of Bio-Environment Control
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    • v.21 no.4
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    • pp.305-311
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    • 2012
  • To investigate the influence electrical conductivity (EC) of nutrient solution and light intensity on growth of red leafy lettuce, fresh and dry weights, number of leave, chlorophyll concentration and production efficiency were evaluated through nutrient film technique system. The levels of EC were 0.5, 1.0, 1.5, 2.0, 3.0, and $6.0dS{\cdot}m^{-1}$, and those of light intensity were 120, 150, and $180{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. Under photoperiod of 16 h/day, the temperature was maintained in the range of $20{\sim}25^{\circ}C$. Planting density was $10{\times}10cm$ (100 plants/$m^2$). When red leafy lettuce were grown in the EC range of $0.5{\sim}1.5dS{\cdot}m^{-1}$, the fresh and dry weights decreased as the EC levels and light intensity were lowered, however, Hunter's a value showed no significant differences among the treatments of EC and light intensity levels (Ex. 1). The fresh and dry weights and production efficiency ($g{\cdot}FW/kw$) were the highest in the treatment of $3.0dS{\cdot}m^{-1}$ and $180{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ when crops were grown under the EC range of EC $1.5{\sim}6.0dS{\cdot}m^{-1}$ (Ex. 2). But the fresh and dry weights, number of leaves, and production efficiency of $2.0dS{\cdot}m^{-1}$ were the highest when the light intensity was $180{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ (Ex. 3). The SPAD value increased gradually as EC levels were elevated. From the above results, we concluded that optimum levels of EC and light intensity were $2.0dS{\cdot}m^{-1}$ and $180{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, respectively, for production as well as production efficiency of red leaf lettuce in plant factory.

Sensory Evaluation and Bioavailability of Red Ginseng Extract(Rg1, Rb1) by Complexation with ${\gamma}$-Cyclodextrin (${\gamma}$-cyclodextrin으로 포접한 홍삼추출물의 관능평가 및 Rg1, Rb1의 생체이용율)

  • Lee, Seung-Hyun;Park, Ji-Ho;Cho, Nam-Suk;Yu, Heui-Jong;You, Sung-Kyun;Cho, Cheong-Weon;Kim, Dong-Chool;Kim, Young-Heui;Kim, Ki-Ho
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.106-110
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    • 2009
  • In order to reduce the bitter taste and improve the bioavailability of red ginseng extract(RGE), inclusion complexes (RGE-CD) of the extract with ${\alpha}-,\;{\beta}-,\;{\gamma}$-cyclodextrin were prepared and studied for their sensory quality and bioavailability compared to RGE. By complexation, the bitter taste-reducing efficacies of ${\alpha}$-CD and ${\beta}$-CD were much lower than that of ${\gamma}$-CD. In comparative sensory analysis for the bitter taste, RGE-${\gamma}$-CD10, prepared using 10%(w/w) of ${\gamma}$-CD, showed a score of 1.93(decreased by about 78%) compared to RGE as the control. In addition, in sensory analysis for flavor, RGE-${\gamma}$-CD10showed a score of 5.60. Upon increasing the amount of ${\gamma}$-CD to 15%(w/w) and 20%(w/w), respectively, the bitter taste of RGE-${\gamma}$-CD was removed and the flavor of RGE disappeared(scores of 2.67 and 1.67, respectively). Therefore RGE-${\gamma}$-CD10 was chosen as an optimum. The same dosages of RGE and RGE-${\gamma}$-CD10 were orally administered to SD(Sprague-Dawley) rats on a saponin basis, and the plasma concentrations of ginsenoside Rg1 and Rb1 were measured over time to estimate the average AUC(area under the plasma concentration versus time curve) of the ginsenosides. After the oral administration, there were no significant differences in the AUC values of the RGE and RGE-${\gamma}$-CD 10 groups for ginsenoside Rg1. However, AUC values for ginsenoside Rb1 were $25.8{\mu}g{\cdot}hr/mL$ in the RGE group and $81.5{\mu}g{\cdot}hr/mL$ in the RGE-${\gamma}$-CD 10 group, respectively. Therefore, the bioavailability of ginsenoside Rb1 in the RGE-${\gamma}$-CD 10 group was significantly higher by up to 315% compared with that in the RGE group(p = 0.0029). These results show that the bitter taste of RGE can be simultaneously removed by the complexation of RGE and ${\gamma}$-CD(RGE-${\gamma}$-CD) along with increased bioavailability.

Spectral Response of $TiO_{2}$/Se : Te Heterojunction for Color Sensor (컬러센서를 위한 $TiO_{2}$/Se : Te 이종접합의 스펙트럼 응답)

  • Woo, Jung-Ok;Park, Wug-Dong;Kim, Ki-Wan;Lee, Wu-Il
    • Journal of Sensor Science and Technology
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    • v.2 no.1
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    • pp.101-108
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    • 1993
  • $TiO_{2}$/Se : Te heterojunction for color sensor has been fabricated by RF reactive sputtering and thermal evaporation methods onto glass substrate. The optimum deposition condition of $TiO_{2}$ films was such that RF power was 120 W, substrate temperature was $100^{\circ}C$, oxygen concentration was 50%, working pressure was 50 mTorr for the $TiO_{2}$ film thickness of $1000{\AA}$. In this case, the optical transmittance of $TiO_{2}$ film at 550 nm-wavelength was 85%, resistivity was $2{\times}10^9{\Omega}{\cdot}cm$, refractive index was 2.3, and optical bandgap was 3.58 eV. The composition ratio of 0 to Ti by AES analysis was 1.7. When $TiO_{2}$ films were annealed at $400^{\circ}C$ for 30 min. in $O_{2}$ ambient, the optical transmittance of $TiO_{2}$ films at the wavelength range of $300{\sim}580$ nm was improved from 0 to 25%. When Se : Te films were annealed at $190^{\circ}C$ for 1 min., photosensitivity under illumination of 1000 lux was 0.75. The optical bandgap of Se : Te films was 1.7 eV. The structures of Se : Te films were the hexagonal with (100) and (110) orientation. The spectral response of a-Se was improved by the addition of Te, especially in the long wavelength region. The $TiO_{2}$/Se : Te heterojunction showed wide spectral response, and more improved one than that of a-Si film in the blue light region.

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Studies on the Induction of Sprouting of Dormant Seed Potato in Fall Crop Production (추작감자의 최아법에 관한 연구)

  • Jae-Young Cho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.21 no.1
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    • pp.97-124
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    • 1976
  • To find out the most effective method of treatments for the induction of sprouting of dormant seed potato pieces for the fall crop production of Irish cobbler, this experiment was carried out with sprout inducing bed and field performance trial. In GA treatments, about 10 days were required to sprout and resulted uniform and thin 3-4 sprouts per tuber piece, but sprouts were slender and rooting was not observed. In Ethrel treatments, 20-25 days were required, and sprouts were inferior than that of GA treatment in uniformity of sprouting, and percentage of rotten pieces and of healthy sprouted tuber pieces, but number of sprouts per tuber pieces was low, being 1-2, and sprouts were short, thick, and healthy, and showed good rooting. In GA and Ethrel mixture treatments, 1-2 more days were required to sprout than GA treatments, but sprouts were relatively healthy, and other sprouting pattern were like that of GA treatments. In Ethylene chlorohydrin and 6-Benzyl-adenine treatments, sprouting was like that of Ethrel treatments, but much more days were required than Ethrel treatments and tendency of severe rotting was observed. Optimum treating methods of promising chemicals found to be 1-2 and 2-5 ppm GA solution, 500 and 1000-2000 ppm Ethrel solution, and 1-2+250-500 and 5+250-500 ppm GA and Ethrel mixture solution for 60 min. treatment of tuber piece and whole-tuber, respectively. Induction of sprouting in dry and hot time resulted severer rotting of tuber pieces during the induction of sprouting and with the advancement of dormancy, being delayed in date of treatment, tendency of promotion of sprouting and rooting was observed. When sprouted tuber piece was transplanted at the same date, yields were in order of Ethrel, GA and Ethrel mixture, and GA treatment, indicating the correlation between yield and healthiness of sprout and rooting status of sprouted tuber piece. In all treatments, earlier transplanting resulted higher yields.

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