Park, Jimin;Kim, Mi-Sun;Joo, Keehyung;Jhon, Gil-Ja;Berry, Edward A.;Lee, Jooyoung;Shin, Dong Hae
Molecules and Cells
/
v.39
no.6
/
pp.495-500
/
2016
We have solved the crystal structure of a predicted fructose-specific enzyme $IIB^{fruc}$ from Escherichia coli ($EcEIIB^{fruc}$) involved in the phosphoenolpyruvate-carbohydrate phosphotransferase system transferring carbohydrates across the cytoplasmic membrane. $EcEIIB^{fruc}$ belongs to a sequence family with more than 5,000 sequence homologues with 25-99% amino-acid sequence identity. It reveals a conventional Rossmann-like ${\alpha}-{\beta}-{\alpha}$ sandwich fold with a unique ${\beta}$-sheet topology. Its C-terminus is longer than its closest relatives and forms an additional ${\beta}$-strand whereas the shorter C-terminus is random coil in the relatives. Interestingly, its core structure is similar to that of enzyme $IIB^{cellobiose}$ from E. coli ($EcIIB^{cel}$) transferring a phosphate moiety. In the active site of the closest $EcEIIB^{fruc}$ homologues, a unique motif CXXGXAHT comprising a P-loop like architecture including a histidine residue is found. The conserved cysteine on this loop may be deprotonated to act as a nucleophile similar to that of $EcIIB^{cel}$. The conserved histidine residue is presumed to bind the negatively charged phosphate. Therefore, we propose that the catalytic mechanism of $EcEIIB^{fruc}$ is similar to that of $EcIIB^{cel}$ transferring phosphoryl moiety to a specific carbohydrate.
Journal of the Korea Academia-Industrial cooperation Society
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v.12
no.2
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pp.1032-1037
/
2011
In this study, simulation and optimization works for a demethanizer column have been performed to obtain ethane and heavier products from a pretreated natural gas stream. Pretreated natural gas feed stream was partially condensed after being precooled by exchanging heat with demethanizer top vapor stream and by using an external refrigeration cycle with a propane refrigerant. Vapor stream was cooled further and partially condensed through a turbo-expander. The power generated from the expansion of turbo-expander was delivered to the compressor for the residue gas compression. Liquid stream was cooled by Joule-Thomson expansion valve and was fed to the middle section of the demethanizer. Recovery percent of ethane for feed natural was set to 80% and methane to ethane molar ratio was fixed as 0.0119. On the other hand, some of the cold heat could be recovered by splitting the feed stream and by exchanging heat with side reboiler in order to reduce the heat duty in the propane refrigeration cycle.
Hyunjin Park;Eunjung Kim;Tae Ho Lee;Sihyun Park;Jang-Duck Choi;Guiim Moon
Food Science of Animal Resources
/
v.43
no.5
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pp.914-937
/
2023
The objective of this study was to establish a multi-residue quantitative method for the analysis of anthelmintic and antiprotozoal drugs in various livestock products (beef, pork, and chicken) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Each compound performed validation at three different levels i.e., 0.5, 1, and 2× the maximum residue limit according to the CODEX guidelines (CAC/GL 71-2009). This study was conducted according to the modified quick, easy, cheap, effective, rugged, and safe procedure. The matrix-matched calibrations gave correlation coefficients >0.98, and the obtained recoveries were in the range of 60.2%-119.9%, with coefficients of variation ≤32.0%. Furthermore, the detection and quantification limits of the method were in the ranges of 0.03-3.2 and 0.1-9.7 ㎍/kg, respectively. Moreover, a survey of residual anthelmintic and antiprotozoal drugs was also carried out in 30 samples of beef, pork, and chicken collected in Korea. Toltrazuril sulfone was detected in all three samples. Thus, our results indicated that the developed method is suitable for determining the anthelmintic and antiprotozoal drug contents in livestock products.
Yun-Juan Bao;Qi Zhou;Xuejing Yu;Xiaolan Yu;Francis J. Castellino
Journal of Microbiology and Biotechnology
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v.33
no.3
/
pp.299-309
/
2023
Glutathione peroxidases (Gpx) are a group of antioxidant enzymes that protect cells or tissues against damage from reactive oxygen species (ROS). The Gpx proteins identified in mammals exhibit high catalytic activity toward glutathione (GSH). In contrast, a variety of non-mammalian Gpx proteins from diverse organisms, including fungi, plants, insects, and rodent parasites, show specificity for thioredoxin (TRX) rather than GSH and are designated as TRX-dependent peroxiredoxins. However, the study of the properties of Gpx in the environmental microbiome or isolated bacteria is limited. In this study, we analyzed the Gpx sequences, identified the characteristics of sequences and structures, and found that the environmental microbiome Gpx proteins should be classified as TRX-dependent, Gpx-like peroxiredoxins. This classification is based on the following three items of evidence: i) the conservation of the peroxidatic Cys residue; ii) the existence and conservation of the resolving Cys residue that forms the disulfide bond with the peroxidatic cysteine; and iii) the absence of dimeric and tetrameric interface domains. The conservation/divergence pattern of all known bacterial Gpx-like proteins in public databases shows that they share common characteristics with that from the environmental microbiome and are also TRX-dependent. Moreover, phylogenetic analysis shows that the bacterial Gpx-like proteins exhibit a star-like radiating phylogenetic structure forming a highly diverse genetic pool of TRX-dependent, Gpx-like peroxidases.
Although Aster yomena has recently attracted attention for its potential anti-cancer, antimicrobial, and immune-boosting effects, there are concerns about residual pesticides because they are consumed in salad-like forms. This study investigated residual characteristics and human dietary risks of the imidazole fungicide prochloraz applied to the herbal medicine Aster yomena with different spray frequency and timing. Residual analysis of prochloraz and its three main metabolites (BTS44595, BTS44596, and 2,4,6-trichlorophenol) in Aster yomena samples was performed using the QuEChERS method and LC-MS/MS. Mean recovery rates of the fungicide and its metabolites were satisfactory in the range of 80.1 ± 1.2% to 108.2 ± 3.8%. The residual concentration of the fungicide calculated as the sum of prochloraz and its metabolites was the highest (4.14 mg·kg-1) in the Aster yomena sample applied three times at weekly intervals with the fungicide until immediately before harvest. The fungicide residue concentration in the Aster yomena was below the method-limit of quantification (MLOQ) when it was applied twice at the interval of 9 day until 21 days before harvest. The theoretical maximum daily intake of prochloraz, calculated based on the daily intake of Aster yomena, mean adult body weight, and the highest residue level analyzed in the this study, was safe at < 80% of the acceptable daily intake of the fungicide (0.01 mg-1·kg·bw-1·day). In conclusion, the triple application method with prochloraz at weekly intervals until the harvest day is recommended to produce safe Aster yomena from the fungicide residues and risks to humans.
Lee, Dong Yeol;Kim, Yeong Jin;Park, Min Ho;Lee, Seung Hwa;Kim, Sang Gon;Kang, Nam Jun;Kang, Kyu Young
The Korean Journal of Pesticide Science
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v.17
no.4
/
pp.307-313
/
2013
This study was carried out to investigate the residual characteristics of fungicide azoxystrobin and difenoconazole in Prunus mume fruits, and establish pre-harvest residue limits (PHRL) based on dissipation and biological half-lives of fungicide residues. The fungicides were sprayed onto the crop at recommended dosage once and 3 times in 7 days interval, respectively. The samples were harvested at 0, 1, 2, 4, 6, 8, 10, 12 and 14 days after treatment. These residual pesticides were extracted with QuEChERS method, clean-up with $NH_2$ SPE cartridge, and residues were analyzed by HPLC/DAD and GLC/ECD, respectively. Method quantitative limits (MQL) of azoxystrobin were 0.03 mg $kg^{-1}$ and of difenoconazole were 0.006 mg $kg^{-1}$. Average recovery were $93.2{\pm}2.49%$, $85.5{\pm}1.97%$ for azoxystrobin at fortification levels at 0.3 and 1.5 mg $kg^{-1}$, and $100.8{\pm}6.74%$, $87.6{\pm}9.92%$ for difenoconazole at fortification levels at 0.06 and 0.3 mg $kg^{-1}$, respectively. The biological half-lives of azoxystrobin were 5.9 and 5.2 days at recommended dosage once and 3 times in 7 days interval, respectively. The biological half-lives of difenoconazole were 9.3 and 8.0 days at recommended dosage once and 3 times in 7 days interval, respectively. The PHRL of azoxystrobin and difenoconazole were recommended as 5.32 and 1.64 mg $kg^{-1}$ for 10 days before harvest, respectively.
In this study, enzymatic saccharification of substrates from Alnus hirsuta Ruper (8-14 years). Quercus acutissima Carruthers, Betula platyphylla var. japonica Nera, Populus euramericana Guiner and Platanus orientalis L. were investigated using crude cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374, and conduced on the optimum treated conditions of the cellulase sacchrification and reactivation of residue of digested substrates. The Trichoderma viride cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. The method of dilignification from wood (5 species) was treated by the peracetic acid(PA) method. The reducing sugar was determined by the dinitrosalicylic acid (DNS) method. 1. The results of tests carried out for 96 hr. (Figure 1), show conclusively the initial substrates from 5 species ($S_3$) which has been rendered highly reactive form and the mean rate of reducing sugar was 28.3 %. 2. The results of tests carried out for 96 hr., the reactivation of residue of digested substrates (improvement in the quality of the substrate through preheating in air at $190^{\circ}C$. for 45 min. followed by milling was (60 mesh size) at the same substrate level, increased concentrations of cellulase at the same substrate level, and increased concentrations of cellulase increases the rate of hydrolysis considerably. 3. Figure 1. shows conclusively that the residue of digested substrates ($S_1$ dried at $60^{\circ}C$) which has been rendered extremly resistant to cellulase action can be reactivated into a highly reactive form ($S_2$), almost comparable to that of the initial substrates ($S_3$). And the reducing sugar formation did not show statistically significent differences at 5% levels by initial substrates and the residue of digested substrates (preheating in air at $190^{\circ}C$. for 45 min. fallowed by milling was (60 mesh size).
Two experiments were conducted to investigate the effects of dietary activated charcoal mixed with wood vinegar (AC) on broiler performance and antibiotics residue in eggs. In experiment 1, a total of low hundred fifty, 1-day-old male broile. chicks (Ross) were divided into 6 groups, consisting 3 replicates of 25 birds each, and 134 one of the six experimental diets: devoid of AC and antibiotics (negative control), devoid of AC with 0.1% antibiotics (positive control), devoid of antibiotics with 1% AC, with 0.1% antibiotics and 1% AC, devoid of antibiotics with 2% AC, with 0.1% antibiotics and 2% AC, for 5 wks. Feed consumption and body weights were measured weekly. At the end of the experiment, eight birds from each group were selected and sacrificed. The relative weight of organs and characteristics of breast muscle were investigated. There were no significant differences in feed intake, body weight gain and feed conversion rate among the treatments. There were also no significant differences in the relative weight of breast meat, leg and liver. The abdominal fat tended to be decreased by the dietary AC. Significant differences were shown (P<0.05) in meat redness (a) and yellowness (b) among the treatments. However, the sensual characteristics of breast muscle was not affected by the dietary treatments. There were significant differences in total microbes, Coli forms and lactic acid bacteria (P<0.05) among the treatments. In experiment 2, a total of ninety, 40-wk-old Hy-line Brown laying hens were divided into 3 groups, consisting 3 replicates of 10 birds each and fed medicated control diet devoid of AC or diets containing 1% AC and 2% AC for 2 wks. The residue of antibiotics in Plasma and egg Yolk were significantly decreased (P<0.05) in 2% AC group compared to that of control. This study suggest that dietary AC may improve the external quality of edible meat and reduced antibiotics residue in chicken eggs without affecting laying performance.
This study was carried out to evaluate residual characteristics of tolclofos-methyl in ginseng and elucidate the reason for its high detection rate from fresh ginseng selling at markets. Seeds of ginseng were sowed after seed dressing with tolclofos-methyl and after a year of growth, the young seedlings were transplanted to field. They were then harvested annually until three-years of growth and the pesticide residue was analyzed in them. LOD and LOQ of the pesticide were 0.001 and 0.003 mg/kg, respectively. Recovery test was carried out to validate the analytical method for tolclofos-methyl in ginseng. The ginseng seedlings were fortified with the test pesticide at the level of LOQ, ten times of LOQ and maximum residue concentration of tolclofos-methyl. Its recovery ranged from 77.37 to 100.16%. Residual concentration of tolclofos-methyl in ginseng seedlings just before transplanting and two-year-old ginseng were from 7.58 to 8.05 and from 6.46 to 6.79 mg/kg, respectively. In case of three-year-old ginseng, it was found to be from 4.18 to 4.35 mg/kg. As a result of annual pesticide residue analysis, concentration of the pesticide was found to decrease time-coursely in ginseng. This may be due to decomposition and increasing of fresh weight of the ginseng during the cultivation periods of three years.
Kim, Yang-Hyeon;Hong, Su-Myeong;Son, Kyung-Ae;Lee, Ju-Young;Min, Zaw Win;Kwon, Hye-Young;Kim, Taek-Kyum;Kyung, Kee-Sung
The Korean Journal of Pesticide Science
/
v.16
no.2
/
pp.121-130
/
2012
In analyzing pesticide residue, LLE (liquid liquid extraction) is generally applied as one of the existing methods, but needed quite a lot of organic solvents and analytical apparatuses for the sample pre-treatment. In addition to its long analysis time and complex analytical processes, it is required to develop a more rapid and efficient method at present. In order to establish an economic and simple pesticide residue analytical method, this study carried out a comparative experiment on the existing analytical method with a new sample pre-treatment method named QuEChERS (quick, easy, cheap, effective, rugged and safe), which extracts and refines pesticide components by directly adding solid powder into the sample. Both the two analytical methods showed favorable values of correlation coefficient ($R^2$ > 0.99) of calibration curves. In terms of the detection limit (identification limit), imidacloprid showed 0.02 mg/kg, while the rest of pesticides showed a level around 0.05 mg/kg. The results of this experiment revealed that the recovery of LLE was 92.8-100.9% and the RSD was below 2.5%. On the other hand, the recovery of QuEChERS was 92.2-101.6% and RSD was below 1.9%. As a result of comparing the amount of pesticide residue by the time between the two analytical methods by using Paired t-Test, there was no significant difference between the two analytical methods as the p-value ranged from 0.3148-0.9890. Considering the results of the two methods, the QuEChERS method had similar recovery, compared to the analytical method using the existing LLE, and the analytical time was shortened by about one fourth of that of the existing method. Moreover, since it excludes the use of harmful organic solvents like dichloromethane during the process of extraction, thus leading to protecting experimenters health and remarkably reducing the amount of disused solvents, it is judged as an echo-friendly and economic analytical method.
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