• Title/Summary/Keyword: P-optimization

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A Study on Optimization for Separation of Phenols and Preconcentration-Separation of Trace Phenols in Reversed-Phase Liquid Chromatography (역상 액체 크로마토그래피에서 페놀류의 분리 최적화 및 미량 페놀류의 농축-분리에 관한 연구)

  • Lee Dai Woon;Lee Sung Won;So, Min Jeong;Cho Byung Yun
    • Journal of the Korean Chemical Society
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    • v.37 no.5
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    • pp.513-522
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    • 1993
  • The purpose of this study is to optimize the selectivity of mobile phase solvents for separation of 25 phenols in reversed phase liquid chromatography and to accomplish the simultaneous preconcentration and separation of trace phenols from water samples. Phenols used in this study were classified into three groups, chloro-, methyl-, and nitrophenols. Quaternary solvent mobile phases were employed to improve the selectivity. Overlapping resolution maps(ORM) as a statistical simplex techniques was used to predict the optimum solvent system. Additional criterion such as pH and temperature were also investigated. In order to improve the resolution and decrease the analysis time, isoselective multisolvent gradient elution system was employed with ORM-Prism method. The simultaneous preconcentration and separation of trace phenols from water samples were performed by using XAD-2/Dowex 1-X8 tandem column. When the extraction efficiency was evaluated by sampling up to 1 L of distilled water, recovery of the phenols, except phenol, was above 90% and the limit of detection of the phenols was 5 ppb. The XAD-2/Dowex 1-X8 method was superior to C18 cartridge in terms of recovery and selectivity.

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Expression of human lactoferrin N-lobe in Pichia pastoris and its antibacterial activity (Pichia pastoris에서 사람 락토페린 N-lobe의 발현과 항균활성)

  • Won, Su-Jin;Jo, Jae-Hyung;Kim, Seung-Hwan;Kwon, Hyuk-Jin;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.271-279
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    • 2015
  • Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

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.

Optimization of Roasted Perilla Leaf Tea Using Response Surface Methodology (반응표면분석을 이용한 들깨잎차 볶음처리의 최적화)

  • Han, Ho-Suk;Park, Jung-Hye;Choi, Hee-Jin;Sung, Tae-Su;Woo, Hi-Seob;Choi, Cheong
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.96-106
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    • 2004
  • Response surface methodology (RSM) was applied in roasting processes of perilla leaves to develop a high quality perilla leaf tea. The Hunter color parameters and electron donating ability were monitored to optimize organoleptic properties of perilla leaf tea. The roasting processes were based on the central composite design with primary variables-roasting temperature $(140{\sim}220^{\circ}C)$, time $(5{\sim}25)$, and reaction variables-sensory test, electron donating ability. From the variables, the roasting condition was optimized using statistical analysis system (SAS) program as developing the functional tea using perilla leaf. Hunter color L and b values of the powdered samples increased with the roasting processes, but Hunter color a value decreased. Electron donating ability was influenced by roasting temperature (p<0.01) and time (p<0.01), and optimum condition selected was at $220^{\circ}C$ for 15 min with coefficient of determinations $(R^2)$ above 0.98. After preference test of perilla leaf tea using parameter of taste, color, and flavor, we can estimate that the optimal roasting condition of preilla leaf for function tea manufacturing are $210{\sim}220^{\circ}C$ for $10{\sim}20$ min by response surface methodology (RSM). Tyrosinase, xanthine oxidase and electron donating ability were 10.14, 14.37 and 59.19% of perilla leaf tea.

Tencel Dyeing by Natural Indigo Prepared from Dyer's Knotweed (천연인디고를 이용한 텐셀직물의 염색)

  • Shin, Youn-Sook;Son, Kyung-Hee;Yoo, Dong-Il
    • Journal of the Korean Society of Clothing and Textiles
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    • v.32 no.12
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    • pp.1963-1970
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    • 2008
  • Natural indigo dye in powder form was prepared by modifying traditional Niram method, using $Ca(OH)_2$ instead of baked oyster powder for precipitating indigo dye. The prepared dye was applied to dyeing Tencel fabrics to investigate the effect of experimental conditions for the optimization of dyeing process. The indigo dye powder contained 15.2%(w/w) of indigo content and 0.757%(w/w) of indirubin content on the basis of HPLC analysis. Maximum dye uptake was obtained at $60^{\circ}C$ for 20min. Almost saturated dye uptake was obtained at 2g/L of sodium hydrosulfite concentration up to 4g/L of indigo dye and then slowly increased for further increase of sodium hydrosulfite. Whereas at higher indigo dye concentration(8g/L) more than 3g/L of reducing agent concentration was required for obtaining the maximum dye uptake. At the same indigo dye and reducing agent concentration, K/S value of the sample dyed without sodium hydroxide(pH 5.75) was 15.19, much higher than one dyed in alkaline condition(K/S 5.76). There was no difference in colorfastness ratings among samples with different color strength. However, more fading was occurred for the sample with low color strength.

Optimization of Dressing Preparation from Yogurt Added Saururus chinensis (Lour.) Bail Extract (삼백초 추출물 첨가 요구르트를 이용한 드레싱 제조의 최적화)

  • HwangBo Mi-Hyang;Kim Hyun-Jeong;Yu Mi-Hee;Lee Ji-Won;Lee In-Sean
    • Korean journal of food and cookery science
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    • v.22 no.1 s.91
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    • pp.22-29
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    • 2006
  • Yogurt base was prepared from milk powder $(14\sim18%)$ to which was added 0.4% Saururus chinensis (Lour.) Bail water extract (SCE) and fermented with lactic acid bacteria (a mixed strain of Streptococcus thermophilus and Lactobacillus bulgaricus) at $37^{\circ}C$ for 24 hr. The viscosity of the yogurt added SCE (SCE yogurt) made from 18% milk was higher than that of SCE yogurt containing $14\sim16%$ milk, whereas the pH and titratable acidity of the SCE yogurt were not significantly different on the range of milk contents. The optimal milk concentration for SCE yogurt manufacture was 18%. In order to optimize the preparation of dressing from SCE yogurt, the central composite design was conducted in terms of the yogurt (30, 40, 50, 60, 70 g), the mayonnaise (6, 12, 18, 24, 30 g) and the salt (0.1, 0.3, 0.5, 0.7, 0.9 g) contents. Sensory evaluation was performed and evaluated using a response surface methodology. The optimum ingredient ratio for SCE yogurt dressing was determined to be 61.2% of yogurt, 23.5% of mayonnaise, 0.58% of salt, 0.58% of honey, 1.75% of mustard, 0.23% of Tabasco pepper sauce, 0.94% of wine and 0.04% of white pepper.

Optimization of Preparation Conditions and Quality Characteristics of Sweet Pumpkin Stock (단호박 스톡 제조조건의 최적화 및 품질 특성)

  • Han, Chi-Won;Park, Won-Jong;Seung, Suk-Kyung
    • Food Science and Preservation
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    • v.15 no.6
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    • pp.832-839
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    • 2008
  • The stock that is the first step for preparation of soups and purees links to the taste of food. Many types of vegetable have been used in stocks, but this study focused on stocks prepared with sweet pumpkin. The stock preparation conditions including the weight of sweet pumpkin, the water volume, and the boiling time at $97^{\circ}C$ were optimized by response surface methodology. The quality characteristics of the resulting stock were investigated. The color, flavor, taste and overall acceptability were dependent parameters. A model equation was proposed with regard to the sweet pumpkin weight, water volume, and boiling time at $97^{\circ}C$. A sweet pumpkin weight of 357.9 to 403.0 g, a water volume of 689.8 to 768.5 mL, and a boiling time of 9.9 to 10.3 min at $97^{\circ}C$ were found to be the optimal stock preparation conditions. The quality characteristics of the sweet pumpkin stock prepared under the optimized conditions were pH 6.64, total acidity 0.18%, soluble solids $2.39\;^{\circ}Brix$, color value (L, 99.07 a, -2.43 b, 11.82), total polyphenol 280.75 mg/L, and electron donating ability 21.32%.

Optimization for Pretreatment Condition according to Salt Concentration and Soaking Time in the Preparation of Perilla Jangachi (소금 농도와 삭힘 시간에 따른 깻잎 장아찌의 전처리 조건의 최적화)

  • Lee, Hye-Ran;Nam, Sang-Min;Lee, Jong-Mee
    • Journal of the Korean Society of Food Culture
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    • v.17 no.1
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    • pp.70-77
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    • 2002
  • Jangachi(salted and fermented vegetable) has been made by Korean traditionally using several kinds of vegetables, which is a good source of variety of nutrients and vitamins. There are many methods for making Jangachi. Generally soy sauce Jangachi is made through two steps. First, as a pretreatment, vegetables are soaked in salt water. Second, soaked vegetables are fermented in various ingredients like soy sauce, sugar, garlic, ginger and so on. This study was performed to observe changes in contents of chemical components and sensory evaluation of pretreated perilla leaf. Perilla leaf was soaked in water with different levels of salt concentration(2, 5 and 8 %) and soaking time(1, 3 and 5 days). The optimal level of salt and soaking time was determined with the results of sensory evaluation by response surface methodology and analysis of composition. The moisture contents decreased as the levels of salt and soaking time increased. The moisture content of untreated sample was 87.5 % and when soaked for 5 days in the water of 8 % salt concentration, it became 78.27 %. pH of Perilla leaf was high in high levels of salt concentration and short soaking time. Total acidity was so opposite to pH that was low in high levels of salt concentration and short soaking time. In the water of 8 % salt concentration, total acidity was 0.14 % when soaked for 1 day, 0.20 % for 3 days and 0.30 % for 5 days. Salt contents became greater as the soaking time increased. As the results of puncture test, soaked Perilla leaf's toughness increased as the levels of salt increased and soaking time decreased. Among the sensory attributes, greenness increased as the levels of salt concentration increased when soaked for more than 3 days. Saltiness and bitterness became greater as the levels of salt concentration increased. Perilla flavor decreased with the short soaking time. Off-flavor increased with the increased levels of soaking time and decreased salt concentration when soaked for more than 3 days. Toughness decreased as the levels of soaking time increased. Crispness increased with the increased levels of salt concentration. The condition of pretreated Perilla was optimum when it soaked for 42 hours in 4 % salt concentration.

Microwave Digestion and Solid-Phase Extraction for Determination of Aluminum in Human Urine by Graphite Furnace Atomic Absorption Spectrometer (흑연로 원자흡수 분광법에 의한 사람 오줌 중 알루미늄의 정량을 위한 마이크로파 삭힘과 고체상 추출)

  • Kim, Young-Sang;Choi, Yoon-Seok
    • Analytical Science and Technology
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    • v.16 no.2
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    • pp.102-109
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    • 2003
  • Microwave digestion and solid-state extraction were studied for determination of trace aluminum{Al(III)} in human urine samples. A mixed acid of nitric acid and hydrogen peroxide was added to urine samples, organic materials were destructed in a home microwave oven and dried in a drying oven. The dried residues were dissolved in a sulfuric acid solution. The solution was eluted through a XAD-4 resin column adsorbed with 8-hydroxyquinoline(Oxine, HQ). Al(III)-8-hydroxyquinolinate complex was formed in the column and eluted with 0.5 M nitric acid solution. The Al(III) eluted was determined by graphite atomic absorption spectrophotometry. Various experimental conditions of followings were investigated for the optimization : the type of acid to dissolve the residues, the amount of HQ adsorbed on the resin, the pH of sample solutions, the type and concentration of acid to elute the complex from column and so on. The contents of Al(III) in real samples were determinated by a calibration curve method. The recovery in standard spiked samples was 94~101% and the detection limit of this procedure was 0.05 ng/mL.

Optimization of Bioflocculant Production Conditions for Organic Wastewater Treatment with Aeromonas hydrophila KH-54 (Aeromonas hydrophila KH-54가 분비하는 유기폐수처리용 생물응집제 생산조건의 최적화)

  • Seo, Ho-Chan;Lee, Jung-Suk;Yun, Zu-Whan;Yi, Yun-Seok;Cho, Hong-Yon
    • Applied Biological Chemistry
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    • v.41 no.6
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    • pp.465-470
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    • 1998
  • For the need of bio-degradable flocculant in stage of wastewater treatment, some cultural conditions of bioflocculant production were optimized with Aeromonas hydrophila KH-54. About 260 strains of type culture and bacteria isolated from marsh, pond, activated sludge, etc were examined for their ability to flocculate kaolin particles and swine wastewater. Among them, KH-54 showed the highest flocculating activity and was identified as Aeromonas hydrophila according to the cultural, morphological and physiological properties. The maximum production of the flocculant secreted by Aeromonas hydrophila KH-54 was observed in culture medium containing 2.0% mannitol, 0.05% ammonium chloride, 0.02% potassium phosphate dibasic, 0.01% $MgSO_4{\cdot}7H_2O$ and 0.05% yeast extract at initial pH 7.0 when cultured on rotary shaker controlled at $25^{\circ}C$ and 150 rpm. Under the optimized condition, the flocculating ability reached to 770 units/ml of kaolin flocculating activity and 81% of NTU removal efficiency against swine wastewater after 4 days cultivation. The bioflocculant was also effective on various organic wastewaters other than swine wastewater, showing NTU removal rate ranging from 92% to 34%.

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