• Title/Summary/Keyword: Rapid enrichment

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Asymmetric Sythesis of Unnatural L-Amino Acids Using Thermophilic Aromatic L-Amino Acid Transaminase

  • Cho, Byung-Kwan;Seo, Joo-Hyun;Kim, Ju-Han;Lee, Chang-Soo;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.299-305
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    • 2006
  • Aromatic L-amino acid transaminase is an enzyme that is able to transfer the amino group from L-glutamate to unnatural aromatic ${\alpha}-keto$ acids to generate ${\alpha}-ketoglutarate$ and unnatural aromatic L-amino acids, respectively. Enrichment culture was used to isolate thermophilic Bacillus sp. T30 expressing this enzyme for use in the synthesis of unnatural L-amino acids. The asymmetric syntheses of L-homophenylalanine and L-phenylglycine resulted in conversion yields of >95% and >93% from 150 mM 2-oxo-4-phenylbutyrate and phenylglyoxylate, respectively, using L-glutamate as an amino donor at $60^{\circ}C$. Synthesized L-homophenylalanine and L-phenylglycine were optically pure (>99% enantiomeric excess) and continuously pre-cipitated in the reaction solution due to their low solubility at the given reaction pH. While the solubility of the ${\alpha}-keto$ acid substrates is dependent on temperature, the solubility of the unnatural L-amino acid products is dependent on the reaction pH. As the solubility difference between substrate and product at the given reaction pH is therefore larger at higher temperature, the thermophilic transaminase was successfully used to shift the reaction equilibrium toward rapid product formation.

Isolation and Characteristics of Listeria monocytogenes from Frozen Foods in Korea (국내에서 판매되는 냉동식품으로부터 Listeria monocytogenes의 분리 및 특성조사)

  • Chang, Yun-Hee
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1324-1329
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    • 1999
  • This study was carried out to investigate the distribution and characteristics of Listeria monocytogenes isolated from frozen Mandoo and pizza in 1998. A total 72 samples were examined and USDA, FDA and modified cold enrichment methods were used for the detection of Listeria spp. Overall prevalence of L. monocytogenes in frozen foods was 9.7% and L. monocytogenes was isolated from 11.1% of frozen Mandoo and 5.6% of frozen pizza. The highest detection rate of Listeria spp. in frozen Mandoo was found at USDA method and the serotype of L. monocytogenes isolates was 4. Isolated L. monocytogenes was confirmed by PCR method with Hly 1 and 2 as primers. It would be necessary to develop more rapid and specific method to isolate and confirm L. monocytogenes from foods because USDA and PCR methods used in this study took 3-4 days. D value of L. monocytogenes isolate in tryptic soy broth was 49.2 sec at $60^{\circ}C$ and 8.8 sec $at\;65^{\circ}C$, and D value of L. monocytogenes in foods with high distribution rate of Listeria spp. would be necessary to evaluate for the safe use of frozen foods.

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Elemental Composition and Source Identification of PM2.5 in Jeju City (제주시 미세먼지(PM2.5)에 함유된 원소의 조성특성 및 오염원)

  • Lee, Ki-Ho;Hu, Chul-Goo
    • Journal of Environmental Science International
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    • v.27 no.7
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    • pp.543-554
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    • 2018
  • From November 2013 to December 2016, ambient fine particulate matter ($PM_{2.5}$) was sampled in the downtown area of Jeju City, South Korea, which has seen rapid urbanization. The atmospheric concentrations of elements were measured in the $PM_{2.5}$ samples. This study focused on Cd, Cr, Cu, Mn, Ni, Pb, As, Sb, Sn, V, and Zn. The concentrations of Al, Na, K, Fe, Ca, Mg, Sr, and La were also obtained for reference. The objectives of this study were to examine the contributions of these elements to $PM_{2.5}$ concentrations in downtown Jeju City, and to investigate the inter-element relationships and the elemental sources by using enrichment factors and principal components analysis (PCA). A composition analysis showed that the 19 elements constituted 6.65 % of the $PM_{2.5}$ mass, and Na, K, Al, Fe, Ca, Mg, and Zn constituted 98 % of the total ion mass. Seasonal trend analysis for the sampling period indicated that the concentrations of the elements increased from November to April. However, no substantial seasonal variations were found in the concentrations of the elements. The composition ratios of some elements (Cu/Zn, Cu/Cd, Cu/Pb, V/Ni, and V/La) were found to be out of range when compared to the literature from other urban areas. The ratios between the elements and the PCA results showed that local contaminant sources in Jeju City rarely influence the composition of $PM_{2.5}$. This suggests that the major sources of $PM_{2.5}$ in Jeju City may include long-range transport of fine particulate matter produced in other areas.

Rapid Screening of Naturally Occurring Radioactive Nuclides (238U, 232Th) in Raw Materials and By-Products Samples Using XRF

  • Park, Ji-Young;Lim, Jong-Myoung;Ji, Young-Yong;Lim, Chung-Sup;Jang, Byung-Uck;Chung, Kun Ho;Lee, Wanno;Kang, Mun-Ja
    • Journal of Radiation Protection and Research
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    • v.41 no.4
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    • pp.359-367
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    • 2016
  • Background: As new legislation has come into force implementing radiation safety management for the use of naturally occurring radioactive materials (NORM), it is necessary to establish a rapid and accurate measurement technique. Measurement of $^{238}U$ and $^{232}Th$ using conventional methods encounter the most significant difficulties for pretreatment (e.g., purification, speciation, and dilution/enrichment) or require time-consuming processes. Therefore, in this study, the applicability of ED-XRF as a non-destructive and rapid screening method was validated for raw materials and by-product samples. Materials and Methods: A series of experiments was conducted to test the applicability for rapid screening of XRF measurement to determine activity of $^{238}U$ and $^{232}Th$ based on certified reference materials (e.g., soil, rock, phosphorus rock, bauxite, zircon, and coal ash) and NORM samples commercially used in Korea. Statistical methods were used to compare the analytical results of ED-XRF to those of certified values of certified reference materials (CRM) and inductively coupled plasma mass spectrometry (ICP-MS). Results and Discussion: Results of the XRF measurement for $^{238}U$ and $^{232}Th$ showed under 20% relative error and standard deviation. The results of the U-test were statistically significant except for the case of U in coal fly ash samples. In addition, analytical results of $^{238}U$ and $^{232}Th$ in the raw material and by-product samples using XRF and the analytical results of those using ICP-MS ($R^2{\geq}0.95$) were consistent with each other. Thus, the analytical results rapidly derived using ED-XRF were fairly reliable. Conclusion: Based on the validation results, it can be concluded that the ED-XRF analysis may be applied to rapid screening of radioactivities ($^{238}U$ and $^{232}Th$) in NORM samples.

Development of Immunochromatography for the Rapid Detection of Listeria monocytogenes (Listeria monocytogenes 신속 검출을 위한 면역크로마토그래피법의 개발)

  • Choi, Jin-Gil;Shim, Won-Bo;Je, Jung-Hyun;Kim, Ji-Young;Lee, Kyu-Ho;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Kim, Kwang-Yup;Kim, Cheol-Ho;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.39 no.3
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    • pp.299-303
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    • 2007
  • The objective of this study was the development of immunochromatography (ICG) for the rapid and accurate detection of Listeria monocytogenes. Here, monoclonal antibodies (MAb) were conjugated with 40 nm colloidal gold particles, where the conjugate was used as the detection reagent in the ICG. The ICG was composed of three pads (sample, conjugate, and absorbance pads) and one nitrocellulose membrane. The colloidal gold-MAb conjugate was applied to the conjugate pad, and the test line and control line on the membrane were treated with MAb (FKLM-3BI2-37) and anti-mouse IgG, respectively. The detection limit of the ICG was $10^{5}$ cell/mL and it showed no cross-reaction to food borne pathogens. We inoculated meat and lettuce samples with various counts of L. monocytogenes, and analyzed them by ICG. All the inoculated meat samples gave positive results after enrichment for 24 h in LEB. These results indicate that ICG was able to serve as a primary screening tool for L. monocytogenes in various foods and agricultural products within 20 min after enrichment.

Selective Detection of Salmonella sp. and Salmonella Typhimurium in Meat by Polymerase Chain Reaction (PCR을 이용한 육류 내 Salmonella sp. 및 Salmonella Typhimurium 분리 검출)

  • Joo, Jong-Won;Hong, Kyung-Pyo;Kim, Yong-Hui;Cho, Sang-Buem
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.2
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    • pp.295-300
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    • 2009
  • The specificity and sensitivity of oligonucleotide primers were examined for the rapid detection of Salmonella in meat samples. The oligonucleotide primers used in this study were designed with the modification of mdh and invA sequence in the chromosome of Salmonella Typhimurium. Through the subsequent analysis of the specificity and sensitivity of the primers, two types of oligonucleotide primers, SLM1 and SLT4 were selected for the detection of Salmonella genus specific and S. Typhimurium species specific, respectively. The lowest detection limit of each primer was represented as 1 cell per reaction when reacted with a prepared DNA solution. The detection efficiency of the two primers was analysed with beef and pork samples intentionally contaminated with a mixture of Salmonella culture, and three preparation methods -, namely direct reaction after extraction, enrichment after extraction, and DNA extraction after enrichment for PCR reaction, - were also compared. No differences were found in the results according to meat sources and preparation methods.

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Comparison of Loop-mediated Isothermal Amplification and Korea Standard Food Codex (KFSC) Method for Detection of Salmonella Typhimurium, Listeria monocytogenes Artificially Inoculated in Yuk-hwe and Yuk-sashimi (육회와 육사시미에 접종된 Salmonella Typhimurium와 Listeria monocytogenes 검출을 위한 Loop-mediated isothermal amplification와 식품공전의 배지 시험법, real-time PCR의 검출 성능 비교)

  • Gwak, Seung-Hae;Lee, So-Young;Kim, Jin-Hee;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.277-282
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    • 2019
  • The object of this study is to compare the performance of the 3M Molecular Detection Assay 2 (3M MDA 2) and the Korea Standard Food Codex (KSFC) Method (i.e., isolation media and real-time PCR) in detecting Salmonella Typhimurium and Listeria monocytogenes in traditional Korean foods. Yuk-hwe and Yuk-sashimi (types of raw beef dishes) were artificially inoculated with $10^0-10^4CFU/25g$ of L. monocytogenes and S. Typhimurium. Citrobacter freundii and Listeria innocua were used as competitive microflora. After enrichment, the samples were analyzed using 3M MDA 2 and real-time PCR. All samples inoculated at concentrations of $10^0-10^4CFU/25g$ without competitive microflora were positive for S. Typhimurium and L. monocytogenes, as detected by 3M MDA 2 and Korea Standard Food Codex (KFSC) Method. In addition, part of the samples were positive for the presence of C. freundii and L. innocua. The 3M MDA 2 - Salmonella and Korea Standard Food Codex (KFSC) Method showed similar detection performances in Yuk-hwe and Yuk-sashimi. The 3M MDA 2 method for Salmonella and Listeria, which is a LAMP-based technology, can be used for rapid detection of S. Typhimurium and L. monocytogenes in raw beef. LAMP bioluminescence assays provide results on the subsequent day and are simple to use compared with the Korea Standard Food Codex (KFSC) Method, particularly in terms of DNA preparation.

High Efficiency Binding Aptamers for a Wide Range of Bacterial Sepsis Agents

  • Graziani, Ana Claudia;Stets, Maria Isabel;Lopes, Ana Luisa Kalb;Schluga, Pedro Henrique Caires;Marton, Soledad;Ferreira, Ieda Mendes;de Andrade, Antero Silva Ribeiro;Krieger, Marco Aurelio;Cardoso, Josiane
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.838-843
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    • 2017
  • Sepsis is a major health problem worldwide, with an extremely high rate of morbidity and mortality, partly due to delayed diagnosis during early disease. Currently, sepsis diagnosis requires bacterial culturing of blood samples over several days, whereas PCR-based molecular diagnosis methods are faster but lack sensitivity. The use of biosensors containing nucleic acid aptamers that bind targets with high affinity and specificity could accelerate sepsis diagnosis. Previously, we used the systematic evolution of ligands by exponential enrichment technique to develop the aptamers Antibac1 and Antibac2, targeting the ubiquitous bacterial peptidoglycan. Here, we show that these aptamers bind to four gram-positive and seven gram-negative bacterial sepsis agents with high binding efficiency. Thus, these aptamers could be used in combination as biological recognition elements in the development of biosensors that are an alternative to rapid bacteria detection, since they could provide culture and amplification-free tests for rapid clinical sepsis diagnosis.

Development of Liposome Immunoassay for Salmonella spp. using Immunomagnetic Separation and Immunoliposome

  • Shin, Jung-Hee;Kim, Myung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1689-1694
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    • 2008
  • The ability to detect Salmonella spp. is essential in the prevention of foodborne illness. This study examined a Salmonella spp. detection method involving the application of immunomagnetic separation and immunoliposomes (IMS/IL) encapsulating sulforhodamine B (SRB), a fluorescent dye. A quantitative assay was conducted by measuring the fluorescence intensity of SRB that was produced from an immunomagnetic bead-Salmonella spp.-immunoliposome complex. The results indicated detection limits of $2.7{\times}10^{5}$ and $5.2{\times}10^{3}$ CFU/ml for Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterka subsp. enterka serovar Typhimurium (S. Typhimurium), respectivley. The signal/noise ratio was improved by using 4% skim milk as a wash solution rather than 2% BSA. In addition, higher fluorescence intensity was obtained by increasing the liposome size. Compared with the conventional plating method, which takes 3-4 days for the isolation and identification of Salmonella spp., the total assay time of to h only including 6 h of culture enrichment was necessary for the Salmonella detection by IMS/IL. These results indicate that the IMS/ IL has great potential as an alternative rapid method for Salmonella detection.

Critical Enhancement of Photothermal Effect by Integrated Nanocomposites of Gold Nanorods and Iron Oxide on Graphene Oxide

  • Yun, Kum-Hee;Seo, Sun-Hwa;Kim, Bo-Mi;Joe, Ara;Han, Hyo-Won;Kim, Jong-Young;Jang, Eue-Soon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2795-2799
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    • 2013
  • Irradiation of gold nanorods (GNRs) with laser light corresponding to the longitudinal surface plasmon oscillation results in rapid conversion of electromagnetic energy into heat, a phenomenon commonly known as the photothermal effect of GNRs. Herein, we propose a facile strategy for increasing the photothermal conversion efficiency of GNRs by integration to form graphene oxide (GO) nanocomposites. Moreover, conjugation of iron oxide (IO) with the GO-GNR nanohybrid allowed magnetic enrichment at a specific target site and the separated GO-IO-GNR assembly was rapidly heated by laser irradiation. The present GO-IO-GNR nanocomposites hold great promise for application in various biomedical fields, including surface enhanced Raman spectroscopy imaging, photoacoustic tomography imaging, magnetic resonance imaging, and photothermal cancer therapy.