• Title/Summary/Keyword: Partial purification

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Technical Trends of Hydrogen Production (수소생산 기술동향)

  • Ryi, Shin-Kun;Han, Jae-Yun;Kim, Chang-Hyun;Lim, Hankwon;Jung, Ho-Young
    • Clean Technology
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    • v.23 no.2
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    • pp.121-132
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    • 2017
  • The increase of greenhouse gases and the concern of global warming instigate the development and spread of renewable energy and hydrogen is considered one of the clean energy sources. Hydrogen is one of the most elements in the earth and exist in the form of fossil fuel, biomass and water. In order to use hydrogen for a clean energy source, the hydrogen production method should be eco-friendly and economic as well. There are two different hydrogen production methods: conventional thermal method using fossil fuel and renewable method using biomass and water. Steam reforming, autothermal reforming, partial oxidation, and gasification (using solid fuel) have been considered for hydrogen production from fossil fuel. When using fossil fuel, carbon dioxide should be separated from hydrogen and captured to be accepted as a clean energy. The amount of hydrogen from biomass is insignificant. In order to occupy noticeable portion in hydrogen industries, biomass conversion, especially, biological method should be sufficiently improved in a process efficiency and a microorganism cultivation. Electrolysis is a mature technology and hydrogen from water is considered the most eco-friendly method in terms of clean energy when the electric power is from renewable sources such as photovoltaic cell, solar heat, and wind power etc.

Partial Purification and Some Properties of Amylases from Germinating Corn(Zea mays L.) (발아 옥수수 amylases의 정제 및 특성)

  • Lee, Tae-Ho;Jung, Tae-Yung;Park, Mi-yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.625-635
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    • 1990
  • The purpose of this study was focused on investigation of biochemical properties of amylases in germinating corn(Zea mays L.) the amylase(I), (II) and (III) from germinating corn seeds were partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-50 ion exchange column chromatography and Sephadex G-100 gel filtration. The last step was effective for separation of the corn amylases to a homogeneous slate. the purified amylase(I) was identified as a kind of $\alpha$-amylase from the fact that 5% starch solution was hydrolysed into mainly maltose and maltotetrose by it, and amylase(II) and amylase(III) were enzymes producing maltotetrose as main product. The molecular weight and specific activity of the amylase(I), (II) and (III) were determined to be 54,000 and 70.47 unit/mg, 39,000 and 62.98 unit/mg, and 51,000 and 80.39 unit/mg, respectively. It showed a tendency to increase the amylases activities in presence of Ba, Ca, Co and Fe groups, but inhibits in that of Ag, Sn, Hg and Zn groups, and amylase(I), (II) and (III) remained stable at pH 5-6 and 2$0^{\circ}C$ for 40 days in containing of 1 mM CaCl$_2$. The optimum pH and optimum temperatures were pH 6, pH 5 and pH 6 and 35$^{\circ}C$, 55$^{\circ}C$ and 55$^{\circ}C$, respectively. These results suggest that the amylase(I), (II) and (III) were different amylases.

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Partial Purification and General Properties of Yeast Acetolactate Synthase (효모 Acetolactate Synthase의 부분 정제와 일반 특성 연구)

  • Koh, Eun-Hie;Song, Soo-Mee;Kim, Sun-Young
    • Journal of the Korean Chemical Society
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    • v.39 no.6
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    • pp.459-465
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    • 1995
  • Acetolactate Synthase (ALS) was partially purified from the yeast and its basic biochemical studies were carried out. Yeast was grown in the minimum media containing 0.5% glucose, 51 mM $K_2HPO_4$, 22 mM $KH_2PO_4$, 8 mM $(NH_4)2SO_4,\;0.4\;m M\;MgSO_4$ for 18 hours at 37 $^{\circ}C$. The cell was ruptured in the buffer (20 mM phosphate buffer pH 7.0, 0.1 mM TPP, 0.5 mM DTT, 1 ${\mu}M$ FAD, and 1 mM MgCl_2$) following an overnight suspension. The supernatant fraction was collected from $10,000{\times}g$ and the enzyme was further purified by ammonium sulfate fractionation, DEAE-Sephacel chromatography and leucine-agarose chromatography. The enzyme activity was measured under the various conditions by the function of protein concentration, time, temperature, pH, and substrate. The optimum temperature was found to be 50$^{\circ}C$, optimum pH 8.0∼8.5. The kinetic parameters, $K_m\;and\;V_{max}$ were 8.4 mM and 17.9 nmol/mg/min respectively. Stability of the enzyme was studied with ethylene glycol and glycerol added to the enzyme solution. Both ethylene glycol and glycerol improved the enzyme stability up to 50%. The study of feedback inhibition showed that valine was a strong inhibitor while leucine was a weak inhibitor.

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Development of Optimal Chlorination Model and Parameter Studies (최적 염소 소독 모형의 개발 및 파라미터 연구)

  • Kim, Joonhyun;Ahn, Sooyoung;Park, Minwoo
    • Journal of Environmental Impact Assessment
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    • v.29 no.6
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    • pp.403-413
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    • 2020
  • A mathematical model comprised with eight simultaneous quasi-linear partial differential equations was suggested to provide optimal chlorination strategy. Upstream weighted finite element method was employed to construct multidimensional numerical code. The code was verified against measured concentrations in three type of reactors. Boundary conditions and reaction rate were calibrated for the sixteen cases of experimental results to regenerate the measured values. Eight reaction rate coefficients were estimated from the modeling result. The reaction rate coefficients were expressed in terms of pH and temperature. Automatic optimal algorithm was invented to estimate the reaction rate coefficients by minimizing the sum of squares of the numerical errors and combined with the model. In order to minimize the concentration of chlorine and pollutants at the final usage sites, a real-time predictive control system is imperative which can predict the water quality variables from the chlorine disinfection process at the water purification plant to the customer by means of a model and operate the disinfection process according to the influent water quality. This model can be used to build such a system in water treatment plants.

Production and Characterization of the Beneficial β-glucuronidase Inhibitor from non pathogenic wild yeast, Candida oleophila WP5-19-1, and its effects on gut microbes (비병원성 야생효모 Candida oleophila WP5-19-1로부터 장내유용세균 유해 효소 β-glucuronidase 저해제의 생산 및 특성)

  • Seon-Jeong Park;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.50 no.1
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    • pp.31-40
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    • 2022
  • We aimed to produce a potent β-glucuronidase inhibitor from wild yeast that could inactivate toxic substances in the colon. Culture supernatants and cell-free extracts of non-pathogenic wild yeasts were prepared and their β-glucuronidase inhibitory activities were measured. Cell-free extract from Candida oleophila WP5-19-1 showed the highest β-glucuronidase inhibitory activity (49.0%). The β-glucuronidase inhibitor was maximally produced (IC50 value; 8.4 mg) when C. oleophila WP5-19-1 was cultured in potato dextrose medium containing 5% dextrose (initial pH; 6.0) at 30℃ for 24 hours. β-glucuronidase inhibitor of C. oleophila WP5-19-1 was partially purified by trypsin hydrolysis, ultrafiltration (3 kDa), and Sephadex G-50 filtration. The partially purified β-glucuronidase inhibitor was stable from 30℃ to 60℃ and at pH 6.0 9.0, and showed residual inhibitory activity of about 80%.

The Development of Squid(Todarodes pacificus) Sik-hae in Kang-Nung District -2. The Effects of Fermentation Temperatures and Periods on Chemical and Microbial Changes, and the Partial Purification of Pretense- (강릉지방의 오징어 식해 개발에 관한 연구 -2. 숙성온도 및 기간에 따른 화학적 변화, 미생물 변화 및 단백질 분해 효소의 정제-)

  • KIM Sang-Moo;CHO Young-Je;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.3
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    • pp.223-231
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    • 1994
  • In order to develop the squid(Todarodes pacificus) sik-hae, the changes of TBA, fatty acids, free amino acids, and the number of microflora fermented at different fermentation temperatures and periods were determined. In addition, pretense from squid sik-hae was partially purified. The number of TBA was the highest after 5-day storage and decreased after that, and lipid oxidation was the highest at $10^{\circ}C$. The amounts of linoleic aid(18:2) and oleic acid (18:1) were about $60\%$ of fatty acid composition of squid sik-hae, and linolenic acid(18:3) and EPA(20:5) significantly decomposed with increasing fermentation periods and temperatures. Pro, His, Aeg, Leu, and Glu were composed mainly of amino acid and the composition ratios of Ser, His, and Arg decreased with increasing fermentation periods whereas, those of Glu, Ala, Val, and Tyr increased. The composition ratios of Glu, Val, and Met increased with increasing fermentation temperatures whereas, those of Ala, Cys, Thr, and Gly decreased. The number of microflora generally increased up to 15-days of storage and decreased after that. The rates of increass and decreass of the microbial number increased in proportion to fermentation temperatures. In addition, the bacteria producing proteases were identified as Bacillus spp. Proteases from $60{\sim}80\%$ ammonium sulfate concentration showed the highest activity and had about 15 binds with molecular weights between 20,000 and 40,000 Dalton by the SDS-PAGE.

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Partial Purification of OsCPK11 from Rice Seedlings and Its Biochemical Characterization (벼 유식물에서 OsCPK11의 부분 정제 및 생화학적 특성 규명)

  • Shin, Jae-Hwa;Kim, Sung-Ha
    • Journal of Life Science
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    • v.30 no.2
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    • pp.137-146
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    • 2020
  • Calcium is one of the important secondary signaling molecules in plant cells. Calcium-dependent protein kinases (CDPK)-the sensor proteins of Ca2+ and phosphorylating enzymes-are the most abundant serine/threonine kinases in plant cells. They convert and transmit signals in response to various stimuli, resulting in specific responses in plants. In rice, 31 CDPK gene families have been identified, which are mainly involved in plant growth and development and are known to play roles in response to various stress conditions. However, little is known about the biochemical characteristics of CDPK proteins. In this study, OsCPK11-a CDPK in rice-was partially purified, and its biochemical characteristics were found. Partially purified OsCPK11 from rice seedlings was obtained by three-step column chromatography that involved anion exchange chromatography consisting of DEAE, hydrophobic interaction chromatography consisting of phenyl-Sepharose, and gel filtration chromatography consisting of Sephacryl-200HR. An in vitro kinase assay using partially purified OsCPK11 was also performed. This partially purified OsCPK11 had a molecular weight of 54 kDa and showed a strong hydrophobic interaction with the hydrophobic resin. In vitro kinase assay showed that the OsCPK11 also had Ca2+-dependent autophosphorylation activity. The OsCPK11 phosphorylated histone III-S, and the optimum pH for its kinase activity was found to be 7.5~8.0. The native OsCPK11 shared several biochemical characteristics with recombinant OsCPK11 studied previously, and both had Ca2+-dependent autophosphorylation activity and favored histone III-S as a substrate for kinase activity, which also had a Ca2+-dependence.

Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Studies on the Enzyme from Arthrobacter luteus Accelerating the Lysis of Yeast Cell Walls -II. Separation of the Factor Accelerating the Lysis of Yeast Cell Walls from the Preparation of Crude Zymolyase and Partial Purification of the Zymolyase with the Sephadex G-75 Gel- (Arthrobacter luteus가 생산(生産)하는 효모세포벽(酵母細胞壁) 용해촉진효소(溶解促進酵素)에 관(關)한 연구(硏究) -제 2 보(第2報) : Crude Zymolyase 표품중(標品中)으로부터 효모(酵母) 세포벽(細胞壁) 용해(溶解) 촉진(促進) 인자(因子)의 분리(分離) 및 Sephadex G-75 Gel에 의한 Zymolyase의 부분(部分) 정제(精製)-)

  • Oh, Hong-Rock;Shimoda, Tadahisa;Funatsu, Masaru
    • Korean Journal of Food Science and Technology
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    • v.12 no.4
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    • pp.254-262
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    • 1980
  • A series of experiment were carried out to separate the factor accelerating the lysis of cell wall of $Saccharomyces\;sak{\acute{e}}$ from the preparation of crude zymolyase obtained from Arthrobacter luteus. An attempt was also made to purify the enzyme which is essential for the study on the separation of the factor. The results are summarized as follows: 1. Crude zymolyase was fractionated 5 peaks $(A{\sim}E)$ containing three peaks $(A{\sim}C)$ passed through the column by the chromatography on Biogel CM-30. 2. Among the five peaks, peak E (protease fraction) was found to contain the factor accelerating the lytic activity of the zymolyase. 3. L-c fraction purified in almost free form from the nonlytic ${\beta}-1$, 3-glucanase, protease and inert protein by the affinity adsorption chromatography with Sephadex G-75 gel was obtained from zymolyase fraction (peak D). When it was subjected to polyacrylamide gel disc electrophoresis, only one clear protein band was observed at pH 4. 5, but still detected two or more band at pH 8. 3.

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Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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