Hae-Yeong Kim

Kyung Hee University, Sŏul, Seoul, South Korea

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Publications (60)118.25 Total impact

  • [show abstract] [hide abstract]
    ABSTRACT: A simple, inexpensive, and universal method to quantify the recombinant proteins in Escherichia coli cell lysate using differential scanning fluorimetry (DSF) is reported. This method is based on the precise correlation between Δ(fluorescence intensity) determined by DSF and the amount of protein in solution. We first demonstrated the effectiveness of the DSF method using two commercially available enzymes, α-amylase and cellobiase, and then confirmed its utility with two recombinant proteins (amylosucrase and maltogenic amylase) expressed in E. coli. The Δ(fluorescence intensity) in DSF analysis accurately correlated with the concentration of the purified enzymes as well as the recombinant proteins in E. coli cell lysates. The main advantage of this method over other techniques such as Western blotting, enzyme linked immunosorbent assay (ELISA), and green fluorescence protein (GFP) fusion proteins is that intact recombinant protein can be quantified without the requirement for additional chemicals or modifications of the recombinant protein. This DSF assay can be performed using widely available equipment such as a RT-PCR instrument, microplates or microtubes and fluorescent dye. This simple but powerful method can be easily applied in a wide range of research areas that require quantification of expressed recombinant proteins.
    Analytical Biochemistry 10/2013; · 2.58 Impact Factor
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    ABSTRACT: Listeriosis is a foodborne disease caused by the pathogenic Listeria monocytogenes and is considered as a serious health problem due to the severity of symptoms and its high mortality rate. Listeria genus is divided into six species and especially L. monocytogenes is an important foodborne pathogen in humans and livestock. Recently, other Listeria species are reported as pathogenic strains in decayed foods and environments as well. High mortality rate of listeriosis demands for rapid methods to detect the potential presence of the food pathogens in the food industry. We have developed a multiplex PCR for rapid and simultaneous detection of six Listeria species including Listeria grayi, Listeria innocua, Listeria ivanovii, L. monocytogenes, Listeria seeligeri and Listeria welshimeri to identify specific Listeria species in processed foods. The optimized multiplex PCR in this study utilized one Listeria genus specific and each Listeria species-specific primer pairs. Each primer pair yields the products of 370-bp for Listeria genus-specific, 201-bp for L. grayi-specific, 749-bp for L. innocua-specific, 463-bp for L. ivanovii-specific, 509-bp for L. monocytogenes-specific, 673-bp for L. seeligeri-specific and 281-bp for L. welshimeri-specific. We have successfully applied multiplex PCR strategy to 93 Listeria isolates from processed meat products to determine specific Listeria species and out of which 81 strains of L. monocytogenes, 10 strains of L. innocua and 2 strains of L. welshimeri were identified. This established multiplex PCR provides rapid and reliable results and will be useful for the detection of Listeria species in contaminated food products and clinical samples.
    Food Control. 08/2013; 32(2):659–664.
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    ABSTRACT: BACKGROUND: The lactic acid bacteria (LAB) in salted Chinese cabbage, the main ingredient of kimchi, were analyzed by culture-dependent SDS-PAGE followed by sequencing of the 16S rRNA gene and by culture-independent PCR-DGGE followed by sequencing of the V3 region of the 16S rRNA gene. The results were compared to those of LAB that had previously been found in kimchi. RESULTS: The two identification methods produced distinct overall LAB profiles. The PCR-DGGE method detected a more diverse microflora, including non-LAB strains. The culture-dependent method uniquely detected Weissella sp. and was able to provide the quantitative distribution of LAB in samples. However, Leuconostoc (Lc) mesenteroides, Lactobacillus curvatus, and Lc carnosum, which had also been reported as the dominant LAB in kimchi in the previous studies, were identified by both methods. CONCLUSION: The two identification methods gave different bacterial profiles while both methods were sufficient to identify the most prevalent LAB in salted Chinese cabbage samples. The quantitative feature of the culture-dependent identification method would make it preferable for studying and monitoring LAB viability in kimchi at each fermentation stage. The availability of the culture-independent identification method to identify a broader bacterial profile, including non-LAB, would make it a more effective tool for controlling contamination of undesirable bacteria during kimchi fermentation.
    Journal of the Science of Food and Agriculture 06/2013; · 1.76 Impact Factor
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    ABSTRACT: 2,3-Butanediol (2,3-BDO) has immense industrial applications. Recently, microbial fermentation has emerged as an alternative way to produce this industrially important chemical. Although (2,3-BDO) is produced by several microorganisms, the Klebsiella genera has an excellent production of 2,3-BDO compared to other 2,3-BDO-producing microorganisms. In order to produce 2,3-BDO on a large scale, the challenges of removing pathogenic factors from Klebsiella pneumoniae need to be addressed. K. pneumoniae produces a number of virulence factors that contribute to its pathogenesis, including lipopolysaccharides, capsules, fimbrial adhesins etc. Removal of these pathogenic factors from 2,3-BDO-producing Klebsiella strains will result in avirulent strains for the safe, economic, and efficient production of 2,3-BDO. In this review, we summarize the current trends in 2,3-BDO production using K. pneumoniae and insights into the removal of its virulence factors for industrial applications.
    Journal of Microbiology and Biotechnology 06/2013; · 1.40 Impact Factor
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    ABSTRACT: Genetically modified (GM) soybean and derived products make up a large part of the biotech-derived food and feed market. As more GM soybean varieties have been approved for commercialization, labeling requirement by South Korea and other countries needs the technical testing methods. This paper reports the development of a multiplex PCR method for identifying six commercialized GM soybean events using the event-specific fragment. Event specific primers targeting Roundup Ready Soybean (RRS, GTS40-3-2), A2704-12, DP356043-5, MON89788, A5547-127, and DP305423-1 were designed, and a multiplex PCR assay consisting of six event-specific fragments and one endogenous lectin fragment was developed. The specificity of the event-specific PCR method was confirmed using 20 GM events of maize, soybean, cotton, and canola. The limit of detection (LOD) for each event in the multiplex PCR is approximately 0.05%. Intra-lab validation by two different operators confirmed the specificity and LOD of this multiplex PCR method. The method was used to test 30 soybean-derived foods from South Korean and US markets, and results revealed three varieties of GM soybean (RRS, A2704-12, and MON89788) in 19 of the 30 food samples tested. This work provides an efficient and cost-effective approach for event-specific analysis of six commercialized GM soybean varieties and related processed foods in Korea.
    Food Control. 06/2013; 31(2):366–371.
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    ABSTRACT: We introduce a nanoporous membrane based impedimetric immunosensor for the label-free detection of bacterial pathogens in whole milk. A simple and rapid method to modify a commercially available alumina nanoporous membrane with hyaluronic acid (HA) effectively reduced the non-specific binding of biomolecules and other cells, and permitted successful immobilization of antibodies. Escherichia coli O157:H7, one of the most harmful food-borne pathogenic bacteria, was tested as a model pathogen in this study. The ionic impedance of electrolytes through nanopores, due to antibody-pathogen interactions, was monitored by impedance spectra and analyzed by normalized impedance change (NIC). The regression equation for the NIC at 1kHz versus concentration of E. coli O157:H7 (10-10(5)cfu/ml) was obtained, and the detection limit found to be as low as 10cfu/ml. In addition, the proposed immunosensor was successfully used for the detection of E. coli O157:H7 in whole milk samples with the detection limit as low as 83.7cfu/ml with 95% probability. The specificity of the immunosensor was also demonstrated using non-target bacteria, including Staphylococcus aureus, Bacillus cereus, and non pathogenic E. coli DH5α. This study shows that a HA-functionalized nanoporous membrane-based impedimetric sensor is capable of detecting pathogenic bacteria in whole milk without any pretreatment. This is a significant step for evaluating the safety of food and environmental samples and other medical diagnostics.
    Biosensors & bioelectronics 01/2013; 44C:210-215. · 5.43 Impact Factor
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    ABSTRACT: Drug delivery technology is emerging as an interdisciplinary science aimed at improving human health. The controlled delivery of pharmacologically active agents to the specific site of action at the therapeutically optimal rate and dose regimen has been a major goal in designing drug delivery systems. Over the past few decades, there has been considerable interest in developing biodegradable drug carriers as effective drug delivery systems. Polymeric materials from natural sources play an important role in controlled release of drug at a particular site. Polyhydroxyalkanoates, due to their origin from natural sources, are given attention as candidates for drug delivery materials. Biodegradable and biocompatible polyhydroxyalkanoates are linear polyesters produced by microorganisms under unbalanced growth conditions, which have emerged as potential polymers for use as biomedical materials for drug delivery due to their unique physiochemical and mechanical properties. This review summarizes many of the key findings in the applications of polyhydroxyalkanoates and polyhydroxyalkanoate nanoparticles for drug delivery system.
    BioMed research international. 01/2013; 2013:581684.
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    ABSTRACT: A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique followed by sequencing of the 16S rDNA fragments eluted from the bands of interest on denaturing gradient gels was used to monitor changes in the bacterial microflora of two commercial kimchi, salted cabbage, and ingredient mix samples during 30 days of fermentation at 4 degrees C and 10 degrees C. Leuconostoc (Lc.) was the dominant lactic acid bacteria (LAB) over Lactobacillus (Lb.) species at 4 degrees C. Weissella confusa was detected in the ingredient mix and also in kimchi samples throughout fermentation in both samples at 4 degrees C and 10 degrees C. Lc. gelidum was detected as the dominant LAB at 4 degrees C in both samples. The temperature affected the LAB profile of kimchi by varing the pH, which was primarily caused by the temperature-dependent competition among different LAB species in kimchi. At 4 degrees C, the sample variations in pH and titratable acidity were more conspicuous owing to the delayed growth of LAB. Temperature affected only initial decreases in pH and initial increases in viable cell counts, but affected both the initial increases and final values of titratable acidity. The initial microflora in the kimchi sample was probably determined by the microflora of the ingredient mix, not by that of the salted cabbage. The microbial distributions in the samples used in this study resembled across the different kimchi samples and the different fermentation temperatures as the numbers of LAB increased and titratable acidity decreased.
    Journal of Microbiology and Biotechnology 01/2013; 23(1):76-84. · 1.40 Impact Factor
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    ABSTRACT: Outbreaks of foodborne diseases associated with Vibrio species such as V. parahaemolyticus, V. vulnificus, and V. cholerae frequently occur in countries having a dietary habit of raw seafood consumption. For rapid identification of different Vibrio species involved in foodborne diseases, whole-cell protein pattern analysis for 13 type strains of 12 Vibrio species was performed using SDS-PAGE analysis. Pathogenic Vibrio species such as V. parahaemolyticus, V. vulnificus, V. cholerae, V. alginolyticus, V. fluvialis, and V. mimicus were included in the 12 Vibrio species used in this study. Each of the 12 Vibrio species showed clearly specific band patterns of its own. Two different strains of V. parahaemolyticus showed two different SDS-PAGE wholecell protein patterns, giving the possibility of categorizing isolated strains in the same V. parahaemolyticus species into two subgroups. The 36 Vibrio isolates collected from sushi restaurants in Busan were all identified as V. parahaemolyticus by comparing their protein patterns with those of Vibrio type strains. The identified isolates were categorized into two different subgroups of V. parahaemolyticus. The whole-cell protein pattern analysis by SDS-PAGE can be used as a specific, rapid, and simple identification method for Vibrio spp. involved in foodborne diseases at the subspecies level.
    Journal of Microbiology and Biotechnology 08/2012; 22(8):1107-12. · 1.40 Impact Factor
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    ABSTRACT: Pyrococcus sp. strain ST04 is a hyperthermophilic, anaerobic, and heterotrophic archaeon isolated from a deep-sea hydrothermal sulfide chimney on the Endeavour Segment of the Juan de Fuca Ridge in the northeastern Pacific Ocean. To further understand the distinct characteristics of this archaeon at the genome level (polysaccharide utilization at high temperature and ATP generation by a Na(+) gradient), the genome of strain ST04 was completely sequenced and analyzed. Here, we present the complete genome sequence analysis results of Pyrococcus sp. ST04 and report the major findings from the genome annotation, with a focus on its saccharolytic and metabolite production potential.
    Journal of bacteriology 08/2012; 194(16):4434-5. · 3.94 Impact Factor
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    ABSTRACT: A new approach to functionalize the surface of hydrophobic nanocarrier through enzymatic polymerization was demonstrated. The effective coupling between the hydrophobic surface of PHB nanoparticle and PHB chain grown from the enzyme fused with a specific ligand provided a simple way of functionalizing nanoparticles with active protein layers in aqueous environment. PHB nanoparticles loaded with model drug molecule, Nile red, were prepared through oil-in-water emulsion solvent evaporation method and the surface of nanoparticles were functionalized with tumor-specific ligand, RGD4C, fused with PHA synthase that drove the coupling reaction. The functionalized PHB nanoparticles showed a specific affinity to MDA-MB 231 breast cancer cells indicating that the tumor-specific ligand, RGD4C, was effectively displayed on the surface of PHB nanoparticles through enzymatic modification and confers targeting capability on the drug carrier.
    Bioorganic & medicinal chemistry letters 05/2011; 21(10):2941-4. · 2.65 Impact Factor
  • Hyungjae Lee, Hae-Yeong Kim
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    ABSTRACT: Antimicrobial peptides exhibit high levels of antimicrobial activity against a broad range of spoilage and pathogenic microorganisms. Compared with bacteriocins produced by lactic acid bacteria, antimicrobial peptides from the genus Bacillus have been relatively less recognized despite their broad antimicrobial spectra. These peptides can be classified into two different groups based on whether they are ribosomally (bacteriocins) or nonribosomally (polymyxins and iturins) synthesized. Because of their broad spectra and high activity, antimicrobial peptides from Bacillus spp. may have great potential for applications in the food, agricultural, and pharmaceutical industries to prevent or control spoilage and pathogenic microorganisms. In this review, we introduce ribosomally synthesized antimicrobial peptides, the lantibiotic bacteriocins produced by members of Bacillus. In addition, the biosynthesis, genetic organization, mode of action, and regulation of subtilin, a well-investigated lantibiotic from Bacillus subtilis, are discussed.
    Journal of Microbiology and Biotechnology 03/2011; 21(3):229-35. · 1.40 Impact Factor
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    ABSTRACT: A simple and sensitive approach for the detection of marker protein, phosphinothricin acetyltransferase, from genetically modified crops was developed based on the colorimetric transition of polydiacetylene (PDA) vesicles in combination with silica microbeads. PDAs have attracted a great deal of interests as a transducing material due to their special features that allow colorimetric response to sensory signals, as well as their inherent simplicity. However, most PDA-based biosensors require additional analytical equipment such as a fluorescence microscope or UV-Vis spectrometer. In this study, we report a new approach to increase the degree of color transition by coupling antibody-conjugated PDA vesicles with silica microbeads in an effort to monitor the results with the unaided eye or simple RGB analysis. By immobilizing PDA vesicles on silica microbeads, we were able to overcome the disadvantages of colloidal PDA-based sensors and increase the degree of colorimetric changes in response to target molecules to a concentration as low as 20 nM. The additional stresses were given to PDA vesicles by antigen-antibody bridging of PDA vesicles coupled with microbeads, resulting in enhanced blue-red color transition. All the results showed that PDA vesicles in conjunction with silica microbeads will be a promising transducing material for the detection of target proteins in diagnostic and biosensing applications.
    Analytical and Bioanalytical Chemistry 03/2011; 400(3):777-85. · 3.66 Impact Factor
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    ABSTRACT: Sucrose phosphorylase, a hexosyltransferase, that is an important enzyme in starch and sucrose metabolisms, reversibly catalyzes the conversion of sucrose and orthophosphate to fructose and α-d-glucose-1-phosphate. A simple assay method for sucrose phosphorylase using 3,5-dinitrosalicylic acid (DNS) was developed. Its effectiveness was compared with that of a previously used NAD method. The results establish that the DNS method is comparable to the NAD method for the assay of sucrose phosphorylase. In particular, analysis of the enzyme activity level of sucrose phosphorylase (SPase) from Bifidobacterium longum SJ32 revealed that the DNS method is not only simple and accurate, but it also is a time-saving method for assaying sucrose phosphorylase activity. Most importantly, the DNS method is stable in broad pH ranges (pH 4–10), whereas the NAD method showed inaccurate profiles in the alkaline pH ranges (pH 8–10). Kinetic studies on SPase from B. longum SJ32 were performed using the simple DNS method developed in this study.
    Food science and biotechnology 01/2011; 20(2):513-518. · 0.70 Impact Factor
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    ABSTRACT: Microfluidic devices for on-chip amplification of DNA from various biological and environmental samples have gained extensive attention over the past decades with many applications including molecular diagnostics of disease, food safety and biological warfare testing. But the integration of sample preparation functions into the chip remains a major hurdle for practical application of the chip-based diagnostic system. We present a PCR-based molecular diagnostic device comprised of a microfabricated chip and a centrifugal force assisted liquid handling tube (CLHT) that is designed to carry out concentration and purification of DNA and subsequent amplification of the target gene in a single chip. The reaction chamber of the chip contains an array of pillar structures to increase the surface area for capturing DNA from a raw sample of macro volume in the presence of kosmotropic agents. The CLHT was designed to provide an effective interface between sample preparation and the microfluidic PCR chip. We have characterized the effect of various fluidic parameters including DNA capture, amplification efficiency and centrifugal pressure generated upon varying sample volume. We also evaluated the performance of this system for quantitative detection of E. coli O157:H7. From the samples containing 10(1) to 10(4) cells per mL, the C(T) value linearly increased from 25.1 to 34.8 with an R(2) value greater than 0.98. With the effectiveness and simplicity of operation, this system will provide an effective interface between macro and micro systems and bridge chip-based molecular diagnosis with practical applications.
    Lab on a Chip 10/2010; 11(2):259-65. · 5.70 Impact Factor
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    ABSTRACT: Many bacteria are involved in fermentation of doenjang and Bacillus species are known to perform significant roles. Although the SDS-PAGE technique has been frequently used for classification and identification of bacteria in various samples, there has been no investigation of the microbial diversity in doenjang. This study aims to investigate the identification and distribution of dominant Bacillus species in doenjang using SDS-PAGE profiles of whole cell proteins and 16S rDNA sequencing. SDS-PAGE of whole cell proteins of the reference Bacillus strains yielded differential banding patterns that could be considered to be highly specific fingerprints. Bacterial strains isolated from doenjang samples were grouped using whole cell protein patterns, which were confirmed by the analysis of 16S rDNA sequencing. B. subtilis was found to be the most dominant strain in most of the samples, and B. licheniformis and B. amyloliquefaciens were less frequently detected. The results obtained in this study showed that a combined identification method, SDS-PAGE patterns of whole cell proteins and subsequent 16S rDNA sequence analysis, could successfully identify Bacillus species isolated from doenjang.
    Journal of Microbiology and Biotechnology 08/2010; 20(8):1210-4. · 1.40 Impact Factor
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    ABSTRACT: We developed an event-specific DNA microarray system to identify 19 genetically modified organisms (GMOs), including two GM soybeans (GTS-40-3-2 and A2704-12), thirteen GM maizes (Bt176, Bt11, MON810, MON863, NK603, GA21, T25, TC1507, Bt10, DAS59122-7, TC6275, MIR604, and LY038), three GM canolas (GT73, MS8xRF3, and T45), and one GM cotton (LLcotton25). The microarray included 27 oligonucleotide probes optimized to identify endogenous reference targets, event-specific targets, screening targets (35S promoter and nos terminator), and an internal target (18S rRNA gene). Thirty-seven maize-containing food products purchased from South Korean and US markets were tested for the presence of GM maize using this microarray system. Thirteen GM maize events were simultaneously detected using multiplex PCR coupled with microarray on a single chip, at a limit of detection of approximately 0.5%. Using the system described here, we detected GM maize in 11 of the 37 food samples tested. These results suggest that an event-specific DNA microarray system can reliably detect GMOs in processed foods.
    Journal of Agricultural and Food Chemistry 05/2010; 58(10):6018-26. · 2.91 Impact Factor
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    ABSTRACT: Rosa damascena has been manufactured as various food products, including tea, in Korea. A new flavonoid glycoside, kaempferol-3-O-beta-D-glucopyranosyl(1-->4)-beta-D-xylopyranoside, named roxyloside A was isolated from the buds of this plant, along with four known compounds, isoquercitrin, afzelin, cyanidin-3-O-beta-glucoside, and quercetin gentiobioside. The chemical structures of these compounds were determined by spectroscopic analyses, including FAB-MS, UV, IR, (1)H and (13)C NMR, DEPT, and 2D NMR (COSY, HSQC, and HMBC). All the isolated compounds except cyanidin-3-O-beta-glucoside exhibited high levels of inhibitory activity against 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase with IC(50) values ranging from 47.1 to 80.6 microM. Cyanidin-3-O-beta-glucoside significantly suppressed angiotensin I-converting enzyme (ACE) activity, with an IC(50) value of 138.8 microM, while the other four compounds were ineffective. These results indicate that R. damascena and its flavonoids may be effective to improve the cardiovascular system.
    Journal of Agricultural and Food Chemistry 01/2010; 58(2):882-6. · 2.91 Impact Factor
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    ABSTRACT: A multiplex polymerase chain reaction (PCR) method was developed to identify and distinguish 3 kinds of stacked genetically modified (GM) maize (MON810× MON863, NK603×MON863, and NK603×MON810× MON863). Four primer pairs, SSIIb JHF/JHR, C3b 5′/TAP1–3′, HS01/cry-CR01, and HS01/CTP164-3′ yielded 101, 129, 194, and 314 bp amplicons, respectively, Using the genomic DNA of the 3 stacked GM maize as templates, 3 or 4 corresponding PCR amplicons were amplified with similar band intensities by the multiplex PCR. The limit of detection (LOD) was approximately 0.5% for 3 kinds of stacked GM maize, using the multiplex PCR. The detection system using multiplex PCR developed in this study may be applicable to monitoring, identifying, and distinguishing not only the stacked GM maizes but also other stacked genetically modified organisms (GMOs). Keywordsgenetically modified (GM) maize-stacked GM event-detection-multiplex polymerase chain reaction-limit of detection
    Food science and biotechnology 01/2010; 19(4):1029-1033. · 0.70 Impact Factor
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    Han-Nah Kim, Jin Lee, Hae-Yeong Kim, Young-Rok Kim
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    ABSTRACT: A new approach to prepare functional polymeric micelles in a one step reaction using the unique catalytic property of PHA synthase is described. A biopolymer-based nanocarrier with cancer targeting capability was successfully synthesized from genetically engineered PHA synthase fused with RGD peptide.
    Chemical Communications 12/2009; · 6.38 Impact Factor

Publication Stats

323 Citations
220 Downloads
4k Views
118.25 Total Impact Points

Institutions

  • 2002–2013
    • Kyung Hee University
      • • Department of Food Science and Technology
      • • College of Life Sciences
      • • Graduate School of Biotechnology
      • • Graduate School of Biotechnology and Plant Metabolism Research Center
      Sŏul, Seoul, South Korea
  • 2012
    • Ewha Womans University
      Sŏul, Seoul, South Korea
  • 2007–2012
    • University of Massachusetts Amherst
      • • Department of Microbiology
      • • Department of Food Science
      Amherst Center, Massachusetts, United States
  • 2010
    • Gachon University
      • College of BioNano Technology
      Seongnam, Gyeonggi, South Korea
  • 2003–2007
    • University of Suwon
      Suigen, Gyeonggi Province, South Korea
    • Seoul National University
      • College of Medicine
      Seoul, Seoul, South Korea
  • 2002–2004
    • Seoul National University Hospital
      • Department of Internal Medicine
      Seoul, Seoul, South Korea