• Title/Summary/Keyword: optimization conditions

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Optimization of a Virus-Induced Gene Silencing System with Soybean yellow common mosaic virus for Gene Function Studies in Soybeans

  • Kim, Kil Hyun;Lim, Seungmo;Kang, Yang Jae;Yoon, Min Young;Nam, Moon;Jun, Tae Hwan;Seo, Min-Jung;Baek, Seong-Bum;Lee, Jeom-Ho;Moon, Jung-Kyung;Lee, Suk-Ha;Lee, Su-Heon;Lim, Hyoun-Sub;Moon, Jae Sun;Park, Chang-Hwan
    • The Plant Pathology Journal
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    • v.32 no.2
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    • pp.112-122
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    • 2016
  • Virus-induced gene silencing (VIGS) is an effective tool for the study of soybean gene function. Successful VIGS depends on the interaction between virus spread and plant growth, which can be influenced by environmental conditions. Recently, we developed a new VIGS system derived from the Soybean yellow common mosaic virus (SYCMV). Here, we investigated several environmental and developmental factors to improve the efficiency of a SYCMV-based VIGS system to optimize the functional analysis of the soybean. Following SYCMV: Glycine max-phytoene desaturase (GmPDS) infiltration, we investigated the effect of photoperiod, inoculation time, concentration of Agrobacterium inoculm, and growth temperature on VIGS efficiency. In addition, the relative expression of GmPDS between non-silenced and silenced plants was measured by qRT-PCR. We found that gene silencing efficiency was highest at a photoperiod of 16/8 h (light/dark) at a growth temperature of approximately $27^{\circ}C$ following syringe infiltration to unrolled unifoliolate leaves in cotyledon stage with a final SYCMV:GmPDS optimal density $(OD)_{600}$ of 2.0. Using this optimized protocol, we achieved high efficiency of GmPDS-silencing in various soybean germplasms including cultivated and wild soybeans. We also confirmed that VIGS occurred in the entire plant, including the root, stem, leaves, and flowers, and could transmit GmPDS to other soybean germplasms via mechanical inoculation. This optimized protocol using a SYCMV-based VIGS system in the soybean should provide a fast and effective method to elucidate gene functions and for use in large-scale screening experiments.

Degradation Characteristics of Non-degradable Dye in Aqueous Solution by Ozonation (고도산화공정인 오존처리에 의한 난분해성 염료 수용액의 분해특성)

  • Hwang, Se-Wook;Park, Jong-Hwan;Lee, Su-Lim;Eom, Ju-Hyun;Ryu, Sung-Ki;Choi, Ik-Won;Kim, Seong-Heon;Kang, Se-Won;Cho, Ju-Sik;Seo, Dong-Cheol
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.58-64
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    • 2020
  • BACKGROUND: Most of the researches on the dye removal using ozonation have been focused on the removal efficiency. However, the research on their removal characteristics and mechanism according to the reaction time has been still insufficient. METHODS AND RESULTS: In this study, the effects of initial pH and dye concentration with reaction time on the degradation characteristics of methyl orange (MO) and methylene blue (MB) by ozonation were evaluated. The degradation efficiency of MB by ozonation increased with increasing pH. On the other hand, the degradation efficiency of MO by ozonation did not show a significant difference with varing pH. The both MO and MB by ozonation were decomposed within 30 min irrespective of the dye concentration, but the decomposition rates of dyes were faster at lower initial dye concentration. The decomposition efficiency of total organic carbon (TOC) in each dye solution by ozonation was low, which was found to be effective for partial decomposition such as decolorization rather than complete degradation of the dye. CONCLUSION: Overall, ozonation was an effective method for removing nondegradable dyes. However, it is necessary to study the optimization of dye degradation under various environmental conditions for ozonation.

Optimization of Catalytic Reaction for Synthesis of 2-Methyl-4-methoxydiphenylamine (2-Methyl-4-methoxydiphenylamine 합성을 위한 촉매반응의 최적화)

  • Cho, Jeong-Woo;Kim, Eun-Seok;Kim, Kiseok;Kim, Seong-Hoon
    • Applied Chemistry for Engineering
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    • v.10 no.2
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    • pp.293-298
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    • 1999
  • Reaction mechanism was elucidated and reaction condition were optimized for the catalytic reaction synthesizing 2-methyl-4-methoxy-diphenylamine (MMDPA) which is an intermediate of Fluoran heat-sensitive dyestuff. Reactants consisted of 2-methyl-4-methoxyaniline (MMA), 3-methyl-4-nitroanisole (MNA), and cyclohexanone, and 5 wt % Pd/C was used as a catalyst. Experiments were run in an open slurry reactor equipped with reflux condenser, and products were analyzed by means of GC/MS and NMR. MMDPA yield of 90 mole % could be obtained after reaction time of 8~10 hours under the optimal reaction conditions comprising the reaction mass composition of MMA : MNA : cyclohexanone = 1 : 2 : 150 based on MMA input of 0.01 gmoles in xylene solvent, reaction temperature of $160^{\circ}C$, and catalyst amount of 0.5 g. It was found that the rate-determining step of overall reaction was dehydrogenation of the intermediate product obtained from condensation of MMA and cyclohexanone. Overall reaction rate and MMDPA yield were enhanced owing to hydrogen transfer reaction by introducing MNA together with MMA in the reaction mass. Excess cyclohexanone in the reaction mass played an important role of promoting the condensation of MMA and cyclohexanone.

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Quantification of 2-Acetyl-1-pyrroline from the Aroma Rice Germplasm by Gas Chromatography (Gas chromatography를 이용한 향미 유전자원의 2-acetyl-1-pyrroline 정량분석)

  • Kim, Jeong-Soon;Park, One-Sung;Ahn, Sang-Nag;Lee, Jung-Ro;Gwag, Jae-Gyun;Kim, Tae-San;Lee, Sok-Young
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.516-521
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    • 2008
  • This study was conducted to optimize the analysis condition and quantify the 2-acetyl-1-pyrroline (2AP) in the brown aroma rice. Extraction effect of the solvent for 2AP was the order of ethanol>acetonitrile>methanol in the range from 30 to $90^{\circ}C$. In the extraction time of 15, 30, 60, and 90 min, the 30 min had the highest 2AP concentration, and it was decreased according to lapse of time. At grinding time, 5 sec resulted in highest 2AP concentration. It was recommended that five sec grinding time, using ethanol, at $90^{\circ}C$ for 30 min was the optimization conditions to quantify the 2AP. Hyangmibyeo2ho and Aranghyangchalbyeo were mild aroma. In the foreign aroma rice, 11 of 19 accessions of Indica types and 2 of 6 accessions of Japonica types were more than mild aroma. Finally, 30 accessions of aroma rice were selected based on their 2AP concentration and agronomic traits.

Stability and Optimization of Crude Protease Extracted from Korean Kiwifruits (국내산 키위에서 추출한 protease 조효소액의 안정성과 최적화에 관한 연구)

  • Kim, Mi-Hyun;Rho, Jeong-Hae;Song, Hyo-Nam
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.554-558
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    • 2010
  • In the study, the protease activity of kiwifruit (Actinidia deliciosa Planch) cultivated in Korea was estimated, with specific examination of proteolytic effects on myofibrilar protein. The crude protease extract of kiwifruit was prepared in two ways; one in which the kiwifruit was homogenized with buffer followed by centrifugation, and the other were the supernatant was precipitated by saturated ammonium sulfate followed by dialysis. The former had 21.23 mM/mL of protease activity, which corresponded to 112.28 mM/g kiwifruit utilized, and the latter had 11.58 mM/mL and 45.80 mM/g of kiwifruit. The crude protease extract of the kiwifruit showed high specificity for casein substrate followed by bovine serum albumin, egg white, collagen, and elastin, in order. The enzyme lost proteolytic activity in acidic conditions such as pH 2-3, and at high temperatures over $60^{\circ}C$. It showed optimal activity in both pH 3.0 and pH 7.5 as well as at $40^{\circ}C$ for casein substrate and at $50^{\circ}C$ for myofibrilar protein substrate. The proteolytic activity toward casein was high with up to 0.5M salt, followed by a sharp decrease beyond this concentration. On the other hand the proteolytic activity for myofibrilar protein decreased steadily with increasing of salt concentration. Kiwifruit has been used as a for meat tenderizer for in home cooking and these results support the its tenderizing effectiveness of kiwifruit especially for Korean style marinating of meat for cooking.

Optimization of solid phase extraction and simultaneous determination of trace anions in concentrated hydrofluoric acid by ion chromatography (불산 중 극미량 음이온 분석을 위한 고상 추출법 및 이온크로마토그래프를 이용한 동시분석법 확립)

  • Yoon, Suk-Hwan;Jo, Dong-ho;Kim, Hyun-Ji;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.29 no.5
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    • pp.219-224
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    • 2016
  • 불산 중 극미량 음이온의 고상추출과 이온크로마토그래프를 이용한 고감도 분석법이 개발되었다. 불산 중 불소이온이 고상에 의해 제거하였고 이어서 음이온 (F, CH3COO, Cl, Br, NO3, PO43−, SO42−)들이 이온크로마토그래프를 이용하여 연속적으로 분리하였다. 고상 추출법에 영향을 주는 각 인자들 (흡착제의 선택, 시료의 부피 및 pH, 용출 용액과 용출용액의 부피)을 결정하였으며 그 결과 흡착제로서 Oasis WAX 컬럼이 가장 우수하였고 1.0 mL의 시료부피, 용출용액으로 50 mM 초산암모늄염 5 mL가 분리능에서 가장 우수하였다. 개발한 방법에 의한 음이온 (Cl, Br, NO3, PO43−, SO42−)들의 방법검출한계는 25 % 불산용액 (w/w) 중에 0.04~0.30 µg/L의 범위를 보였고 정밀도는 20.0와 40.0 µg/L의 농도에서 5 % 이내를 보였다. 한 제조회사에 의한 25 % 불산 중 음이온의 4.2에서 47.5 µg/L의 범위로 모두 검출되었다. 이 방법은 시험절차가 간단하고, 재현성 및 감도가 좋아서 반도체회사에서 불산 중 음이온 불순물을 정도 관리하는데 매우 유용한 방법이 될 것으로 판단된다.

Computer Simulation of the Effect of Pressurized/Depressurized Distillation Process on the Reduction of Separation Energy of Ethanol from Alcohol Fermented Broth (가압/감압 증류 공정이 발효 알콜의 분리 에너지 절감 효과에 미치는 영향에 관한 전산 모사)

  • 허병기;배천순;김휘동
    • Journal of Energy Engineering
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    • v.2 no.1
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    • pp.123-132
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    • 1993
  • This work is focussed on the reduction of ethanol separation energy from alcohol fermented broth and categorized into the development of a computer program for the design of the pressurized/depressurized distillation process which has been regarded as one of the energy-reducing models for the conventional distillation process, the optimization of operating conditions of distillation towers by means of the developed program, and the evaluation of the total annual energy cost of pressurized/depressurized distillation columns compared with that of the conventional single distillation columns. The operating pressures are, in case of pressurized/depressurized distillation, 3103/760 mmHg, 3103/450 mmHg, 3103/160 mmHg, and in case of conventional distillation, 760 mmHg. The optimum reflex rations which the sum of the annual energy cost and the annual fixed cost for each process becomes minimum are 3.7475/2.9111 for the operating pressures of 3103/760 mmHg, 3.814/2.9712 for 3103/450 mmHg, 3.0783/2.2400 for 3103/150 mmHg, and 3.8544 for the atmospheric operating pressure. And the annual energy cost of pressurized/depressurized distillation process for the above-mentioned operating pressures is distributed between 42% and 47% of that of conventional distillation process.

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Optimization of Production Yield for Neohesperidin by Response Surface Methodology (반응표면 분석법을 이용한 neohesperidin 생산 수율의 최적화)

  • Yang, Hee-Jong;Jeong, Seong-Yeop;Choi, Nack-Shick;Ahn, Keug-Hyun;Park, Chan-Sun;Yoon, Byoung-Dae;Ryu, Yeon-Woo;Ahn, Soon-Cheol;Kim, Min-Soo
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1691-1696
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    • 2010
  • Neohesperidin is a natural new nutrition sweetener, widely existing in plants of dry citrus peel, which can be derived from extraction. Since the sweetness is 1,300-1,500 times greater than that of sugar, neohesperidin are widely used in fruit juices, wines, beverages, bakeries and pharmaceutical formulations, and are particularly suitable for consumption by diabetic patients. However, the yield of extraction from citrus peel waste is very low. In this study optimal yield conditions were determinedusing response surface methodology (RSM) in order to increase the neohesperidin extraction yield. The critical factors for maximum extraction yield were selected extraction pressure ($x_1$), extraction time ($x_2$), and concentration of ethanol ($x_3$). As a result, the extraction yield was improved when the extracting pressure increased. The extraction yield also increased in a time-dependent manner. When adding ethanol as an assistance solvent to the supercritical carbon dioxide, extraction yield was increased as more ethanol concentration was added. Finally, the extraction yield of neohesperidin was improved to about 162.22% compared to ethanol extraction as a conventional method.

Production of gamma-Aminobutyric Acid (GABA) by Lactobacillus plantarum subsp. plantarum B-134 Isolated from Makgeolli, Traditional Korean Rice Wine (한국전통주인 막걸리로부터 분리한 Lactobacillus plantarum subsp. plantarum B-134의 gamma-aminobutyric acid (GABA)의 생산)

  • Lee, Hyun-Ju;Son, Jae-Young;Lee, Sang-Jae;Lee, Han-Seung;Lee, Bae-Jin;Choi, In-Soon;Sohn, Jae Hak
    • Journal of Life Science
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    • v.27 no.5
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    • pp.567-574
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    • 2017
  • This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.

Optimization of Ammonia Percolation Process for Ethanol Production from Miscanthus Sinensis (억새를 이용한 바이오 에탄올 생산을 위한 암모니아 침출 공정 최적화)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.704-711
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    • 2010
  • Lignocellulose ($2^{nd}$ generation) is difficult to hydrolyze due to the presence of lignin and the technology developed for cellulose fermentation to ethanol is not yet economically viable. However, recent advances in the extremely new field of biotechnology for the ethanol production are making it possible to use of agriculture residuals and nonedible crops biomass, e.q., rice straw and miscanthus sinensis, because of their several superior aspects as agriculture residual and nonedible crops biomass; low lignin, high contents of carbohydrates. In this article, as the basic study of AP(Ammonia Percolation), the properties and the optium conditions of process were established, and then the overall efficiency of AP was investigated. The important independent variables for AP process were selected as ammonia concentration, reaction temperature, and reaction time. The percolation condition for maximizing the content of cellulose, the enzymatic digestibility, and the lignin removal was optimized using RSM(Response Surface Methodology). The determined optimum condition is ammonia concentration; 11.27%, reaction temperature; $157.75^{\circ}C$, and reaction time; 10.01 min. The satisfying results were obtained under this optimized condition, that is, the results are as follows: cellulose content(relative); 39.98%, lignin content(relative); 8.01%, and enzymatic digestibility; 85.89%.