• Title/Summary/Keyword: protein recovery

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Optimization of Solid Phase Extraction Method for Quantitative Analysis of Perfluorooctanoic Acid in Serum using Liquid Chromatography-Tandem Mass Spectrometry

  • Lee, Hyun-Seok;Yim, Yong-Hyeon
    • Mass Spectrometry Letters
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    • v.5 no.1
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    • pp.30-33
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    • 2014
  • A solid phase extraction (SPE) method was optimized for the quantitative analysis of perfluorooctanoic acid (PFOA) in serum using hydrophilic-lipophilic balance SPE and LC-MS/MS. Fetal bovine serums spiked with $^{13}C_8$-PFOA before or after SPE were used as test samples for evaluation of the SPE efficiency. Simultaneous evaluation of matrix effects and absolute SPE recovery for $^{13}C_8$-PFOA in serum using different sample pre-treatments and SPE conditions allowed optimization of SPE process efficiency with minimal matrix effect and decent SPE recovery. Introduction of protein precipitation as a sample pre-treatment procedure for serum samples before SPE generally decreased matrix effect in LC-MS/MS analysis and provided more stable recovery of PFOA.

Managing Within-Field Spatial Yield Variation of Rice by Site-Specific Prescription of Panicle Nitrogen Fertilizer

  • Ahn Nguyen Tuan;Shin Jin Chul;Lee Byun-Woo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.4
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    • pp.238-246
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    • 2005
  • Rice yield and protein content have been shown to be highly variable across paddy fields. In order to characterize this spatial variability of rice within a field, two-year experiments were conducted in 2002 and 2003 in a large-scale rice field of $6,600m^2$ In year 2004, an experiment was conducted to know if variable rate treatment (VRT) of N fertilizer, that was prescribed for site-specific management at panicle initiation stage, could reduce spatial variation in yield and protein content of rice while increasing yield compared to conventional uniform N topdressing (UN, 33kg N/ha at PIS) method. VRT nitrogen prescription for each grid was calculated based on the nitrogen (N) uptake (from panicle initiation to harvest) required for target rice protein content of $6.8\%$, natural soil N supply, and recovery of top-dressed N fertilizer. The required N uptake for target rice protein content was calculated from the equations to predict rice yield and protein content from plant growth parameters at panicle initiation stage (PIS) and N uptake from PIS to harvest. This model· equations were developed from the data obtained from the previous two-year experiments. The plant growth parameters for the calculation of the required N were predicted non-destructively by canopy reflectance measurement. Soil N supply for each grid was obtained from the experiment of year 2003, and N recovery was assumed to be $60\%$ according to the previous reports. The prescribed VRT N ranged from 0 to 110kg N/ha with an average of 57kg/ha that was higher than 33 kg/ha of UN. The results showed that VRT application successfully worked not only to reduce spatial variability of rice yield and protein content but also to increase rough rice yield by 960kg/ha. The coefficient of variation (CV) for rice yield and protein content was reduced significantly to $8.1\%$ and $7.1\%$ in VRT from $14.6\%$ and $13.0\%$ in UN, respectively. And also the average protein content of milled rice in VRT showed very similar value of target protein content of $6.8\%$. In conclusion the procedure used in this paper was believed to be reliable and promising method for reducing within-field spatial variability of rice yield and protein content. However, inexpensive, reliable, and fast estimation methods of natural N supply and plant growth and nutrition status should be prepared before this method could be practically used for site-specific crop management in large-scale rice field.

SPATIAL YIELD VARIABILITY AND SITE-SPECIFIC NITROGEN PRESCRIPTION FOR THE IMPROVED YIELD AND GRAIN QUALITY OF RICE

  • Lee Byun-Woo;Nguyen Tuan Ahn
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2005.08a
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    • pp.57-74
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    • 2005
  • Rice yield and protein content have been shown to be highly variable across paddy fields. In order to characterize this spatial variability of rice within a field, the two-year experiments were conducted in 2002 and 2003 in a large-scale rice field of $6,600m^2$ In year 2004, an experiment was conducted to know if prescribed N for site-specific fertilizer management at panicle initiation stage (VRT) could reduce spatial variation in yield and protein content of rice while increasing yield compared to conventional uniform N topdressing (UN, ,33 kg N/ha at PIS) method. The trial field was subdivided into two parts and each part was subjected to UN and VRT treatment. Each part was schematically divided in $10\times10m$ grids for growth and yield measurement or VRT treatment. VRT nitrogen prescription for each grid was calculated based on the nitrogen (N) uptake (from panicle initiation to harvest) required for target rice protein content of $6.8\%$, natural soil N supply, and recovery of top-dressed N fertilizer. The required N uptake for target rice protein content was calculated from the equations to predict rice yield and protein content from plant growth parameters at panicle initiation stage (PIS) and N uptake from PIS to harvest. This model equations were developed from the data obtained from the previous two-year experiments. The plant growth parameters for this calculation were predicted non-destructively by canopy reflectance measurement. Soil N supply for each grid was obtained from the experiment of year 2003, and N recovery was assumed to be $60\%$ according to the previous reports. The prescribed VRT N ranged from 0 to 110kg N/ha with average of 57kg/ha that was higher than 33kg/ha of UN. The results showed that VRT application successfully worked not only to reduce spatial variability of rice yield and protein content but also to increase rough rice yield by 960kg/ha. The coefficient of variation (CV) for rice yield and protein content was reduced significantly to $8.1\%\;and\;7.1\%$ in VRT from $14.6\%\;and\;13.0\%$ in UN, respectively. And also the average protein content of milled rice in VRT showed very similar value of target protein content of $6.8\%$. Although N use efficiency of VRT compared to UN was not quantified due to lack of no N control treatment, the procedure used in this paper for VRT estimation was believed to be reliable and promising method for managing within-field spatial variability of yield and protein content. The method should be received further study before it could be practically used for site-specific crop management in large-scale rice field.

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Kinetic analysis of Drosophila Vnd protein containing homeodomain with its target sequence

  • Yoo, Si-Uk
    • BMB Reports
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    • v.43 no.6
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    • pp.407-412
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    • 2010
  • Homeodomain (HD) is a highly conserved DNA-binding domain composed of helix-turn-helix motif. Drosophila Vnd (Ventral nervous system defective) containing HD acts as a regulator to either enhance or suppress gene expression upon binding to its target sequence. In this study, kinetic analysis of Vnd binding to DNA was performed. The result demonstrates that DNA-binding affinity of the recombinant protein containing HD and NK2-specific domain (NK2-SD) was higher than that of the full-length Vnd. To access whether phosphorylation sites within HD and NK2-SD affect the interaction of the protein with the target sequence, alanine substitutions were introduced. The result shows that S631A mutation within NK2-SD does not contribute significantly to the DNA-binding affinity. However, S571A and T600A mutations within HD showed lower affinity for DNA binding. In addition, DNA-binding analysis using embryonic nuclear protein also demonstrates that Vnd interacts with other nuclear proteins, suggesting the existence of Vnd as a complex.

Purification of Cyclodextrin Glucanotransferase by Affinity Chromatography (Affinity Chromatography를 이용한 Cyclodextrin Glucanotransferase의 정제)

  • 안중훈;황진봉;김승호;김경은
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.313-314
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    • 1991
  • - The cyclodextrin glucanotransferase (CGTase) of a mutant of Bacillus stearothermophilus was purified in one step by affinity chromatography. The recovery was 95%. The specific activity of the CGTase increased from 26.2 U/mg protein to 485.5 U/mg protein. The purified CGTase was almost homogeneous by SDS-polyacrylamide gel electrophoresis. The one-step purification proved to be feasible with the mutant in contrast to the parent strain, which required pre-purification step of ammonium sulfate precipitation.

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Submerged Monoxenic Culture Medium Development for Heterorhabditis bacteriophora and its Symbiotic Bacterium Photorhabdus luminescens: Protein Sources

  • Cho, Chun-Hwi;Whang, Kyung-Sook;Gaugler, Randy;Yoo, Sun-Kyun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.869-873
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    • 2011
  • Most medium formulations for improving culture of entomopathogenic nematodes (EPN) based on protein sources have used enriched media like animal feed such as dried egg yolk, lactalbumin, and liver extract, among other ingredients. Most results, however, showed unstable yields and longer production time. Many of the results do not show the detailed parameters of fermentation. Soy flour, cotton seed flour, corn gluten meal, casein powder, soytone, peptone, casein hydrolysates, and lactalbumin hydrolysate as protein sources were tested to determine the source to support optimal symbiotic bacteria and nematode growth. The protein hydrolysates selected did not improve bacterial cell mass compared with the yeast extract control, but soy flour was the best, showing 75.1% recovery and producing more bacterial cell number ($1.4{\times}10^9$/ml) than all other sources. The highest yield ($1.85{\times}10^5$ IJs/ml), yield coefficient ($1.67{\times}10^6$ IJs/g medium), and productivity ($1.32{\times}10^7$ IJs/l/day) were also achieved at enriched medium with soybean protein.

Thermotolerance Inhibits Various Stress-induced Apoptosis in NIH3T3 Cells

  • Park, Jun-Eui;Lee, Kong-Joo;Kim, Choon-Mi
    • Archives of Pharmacal Research
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    • v.21 no.1
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    • pp.46-53
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    • 1998
  • When NIH3T3 cells were exposed to mild heat and recovered at $37^{\circ}C$ for various time intervals, they were thermotolerant and resistant to subsequent stresses including heat, oxidative stresses, and antitumor drug methotrexate which are apoptotic inducers. The induction kinetics of apoptosis by stresses were determined by DNA fragmentation and protein synthesis using $[35^S]$methionine pulse labeling. We investigated the hypothesis that thermotolerant cells were resistant to apoptotic cell death compared to control cells when both cells were exposed to various stresses inducing apoptosis. The cellular changes in thermotolerant cells were examined to determine which components are involved in this resistance. At first, the degree of resistance correlates with the extent of heat shock protein synthesis which were varied depending on the heating times at $45^{\circ}C$ and recovery times at $37^{\circ}C$after heat shock. Secondly, membrane permeability change was observed in thermotolerant cells. When cells prelabeled with $[^{3}H]$thymidine were exposed to various amounts of heat and recovered at $37^{\circ}C$ for 1/2 to 24 h, the permeability of cytosolic $[^{3}H]$thymidine in thermotolerant cells was 4 fold higher than that in control cells. Thirdly, the protein synthesis rates in thermotolerant and control cells were measured after exposing the cells to the same extent of stress. It turned out that thermotolerant cells were less damaged to same amount of stress than control cells, although the recovery rates are very similar to each other. These results demonstrate that an increase of heat shock proteins and membrane changes in thermotolerant cells may protect the cells from the stresses and increase the resistance to apoptotic cell death, even though the exact mechanism should be further studied.

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Expression of the Heat Shock Protein Genes in Response to Thermal Stress in the Silkworm Bombyx mori

  • Velu, Dhanikachalam;Ponnuvel, Kangayam. M.;Qadri, Syed. M. Hussaini
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.1
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    • pp.21-27
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    • 2008
  • The expression of heat shock protein genes (Hsp 70, Hsp 40, Hsp 20.8 and Hsp 20.4) against thermal stress in silkworm Bombyx mori was performed through semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Upon exposure of silkworm to two temperature regimes ($38^{\circ}C$ and $42^{\circ}C$), significant change in the expression of Hsp gene was observed as compared to the control. Hsp 70 and Hsp 40 showed increased expression than the small heat shock protein genes Hsp 20.8 and Hsp 20.4. The Hsp 70 showed increased expression during the recovery period as compared to 1 hr thermal treatments ($38^{\circ}C$/1 hr and $42^{\circ}C$/1 hr). Whereas, Hsp 40, Hsp 20.8 and Hsp 20.4 genes showed higher expression level at initial stages that later gradually decrease during recovery period. Tissue specific expression of Hsp 70 showed variation in the level of expression amongst the tissues. The mid gut and fat body tissues showed higher expression than the cuticle and silk gland tissue. The Hsp 70, Hsp 40 gene expression was analyzed in thermotolerant (Nistari) and thermo susceptible silk worm strain (NB4D2) and results showed significant variation in their expression level. The Nistari showed higher expression of Hsp 70 and Hsp 40 genes than the NB4D2. These findings provide a better understanding of cellular protection mechanisms against environmental stress such as heat shock, as these Hsps are involved in an organism thermotolerance.

Optimization of the High-Pressure Condition for Rice Protein Extracting Using Response Surface Methodology (RSM) (반응표면분석법을 이용한 쌀 단백질 초고압 추출조건 최적화)

  • Ra, Ha-Na;Park, Sa-Ra;Kim, Ha-Yun;Cho, Yong-Sik;Kim, Kyung-Mi
    • Journal of the Korean Society of Food Culture
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    • v.34 no.6
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    • pp.779-784
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    • 2019
  • The purpose of this study was to optimize the rice protein extracted using a response surface methodology. The experiment was designed based on a CCD (Central Composite Design), and the independent variables were the high pressure (X1, 0-400 MPa) and processing time (X2, 0-10 minutes). The results of the extraction content (Y1), residue content (Y2), and recovery yield (Y3) were fitted to a response surface methodology model (R2= 0.92, 0.92, and 0.93, respectively). Increasing the pressure and processing time has a positive effect on the extraction content (Y1), residue content (Y2), and recovery yield (Y3). Therefore, these high-pressure conditions (independent variables) can significantly affect the improvement in rice protein extraction efficiency. Thus, the optimal conditions of X1 and X2 were 400 MPa and 10 min., respectively. Under these optimal conditions, the predicted values of Y1, Y2, and Y3 were 62.93, 57.53 mg/g, and 91.76%, respectively.

Development of Drink from Composition with Medicinal Plants and Evaluation of Its Physiological Function (생약재를 이용한 음료의 개발 및 기능성 평가)

  • 박성혜;황호선;한종현
    • Journal of Nutrition and Health
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    • v.37 no.5
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    • pp.364-372
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    • 2004
  • The purpose of this study was to investigate the possibility of medicinal plants application as an edible functional food resource. We carried out to develop a traditional functional beverage by using hot-water extraction of 4 medicinal plants (Polygonatun sibiricum, Ophiopogonis radix, Lycii fructus, Schizandriae frutcus) and we examined the effects of drink on physiological response during exercise and recovery phase. The subjects were male baseball players, and exercise protocol was performed with 45 minutes treadmill running and 85% VO$_2$ max intensity. Brix, pH and titratable acidity of developed drink were 9.5%, 3.3 and 0.22%. The approximate nutritional composition of beverage was carbohydrate, 5.98%, total dietary fiber, 0.42%, crude protein, 0.70%, crude fat, 0.20% and crude ash, 0.20%. Developed. traditional functional beverage contained K (4.00 mg%), Na (3.68mg%), Ca (2.54mg%), Mg (1.60mg%) and Fe (0.29mg%). Developed beverage drinking group showed a lowest heart rate during exercise and recovery phase than the other two group (water group and ion beverage group). In the change of blood lactate concentration, developed beverage intake group showed a significant lowest values during exercise and recovery phase. And blood hematocrit values and osmolarity were lowest in the developed beverage intake group. In these results, exercise performance capacity was expected to improve most highly with the developed drink from composition with medicinal plants. Also developed beverage was effective in recovery of exercise-induced fatigue. Thus developed drink with medicinal plants can be used as a functional material improving decrease fatigue effects in beverage industry.