• Title/Summary/Keyword: Bio-analysis

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Application of Denaturing Gradient Gel Electrophoresis to Estimate the Diversity of Commensal Thermophiles

  • Bae, Jin-Woo;Kim, Joong-Jae;Jeon, Che-Ok;Kim, Kwang;Song, Jae-Jun;Lee, Seung-Goo;Poo, Har-Young;Jung, Chang-Min;Park, Yong-Ha;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1008-1012
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    • 2003
  • Symbiobacterium toebii has been reported as a thermophile exhibiting a commensal interaction with Geobacillus toebii. The distribution of the commensal thermophiles in various soils was investigated using a denaturing gradient gel electrophoresis (DGGE). Based on the DGGE analysis, the enrichment condition for the growth of Symbiobacterium sp. was found to also enrich populations of several other microbial spp. as well as Symbiobacterium sp. In the enrichment experiment, several different 16S rDNA sequences of commensal thermophiles were detected in all of the soil samples tested, indicating that commensal thermophiles are widely distributed in various soils.

Identification of Catalytic Amino Acid Residues by Chemical Modification in Dextranase

  • Ko, Jin-A;Nam, Seung-Hee;Kim, Doman;Lee, Jun-Ho;Kim, Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.837-845
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    • 2016
  • A novel endodextranase isolated from Paenibacillus sp. was found to produce isomaltotetraose and small amounts of cycloisomaltooligosaccharides with a degree of polymerization of 7-14 from dextran. To determine the active site, the enzyme was modified with 1-ethyl-3-[3-(dimethylamino)-propyl]-carbodiimide (EDC) and α-epoxyalkyl α-glucosides (EAGs), an affinity labeling reagent. The inactivation followed pseudo first-order kinetics. Kinetic analysis and chemical modification using EDC and EAGs indicated that carboxyl groups are essential for the enzymatic activity. Three Asp and one Glu residues were identified as candidate catalytic amino acids, since these residues are completely conserved across the GH family of 66 enzymes. Replacement of Asp189, Asp340, or Glu412 completely abolished the enzyme activity, indicating that these residues are essential for catalytic activity.

Spore Display Using Bacillus thuringiensis Exosporium Protein InhA

  • Park, Tae-Jung;Choi, Soo-Keun;Jung, Heung-Chae;Lee, Sang-Yup;Pan, Jae-Gu
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.495-501
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    • 2009
  • A new spore display method is presented that enables recombinant proteins to be displayed on the surface of Bacillus spores via fusion with InhA, an exosporium component of Bacillus thuringiensis. The green fluorescent protein and $\beta$-galactosidase as model proteins were fused to the C-terminal region of InhA, respectively. The surface expression of the proteins on the spores was confirmed by flow cytometry, confocal laser scanning microscopy, measurement of the enzyme activity, and an immunogold electron microscopy analysis. InhA-mediated anchoring of foreign proteins in the exosporium of Bacillus spores can provide a new method of microbial display, thereby broadening the potential for novel applications of microbial display.

Correlation Model between Questionnaires of Oriental Medicine and Bio-signal (한방 설문과 생체신호 간 연관분석 모델)

  • Park, Sang Chan;Lee, Sang Chul;Park, Young Bae;Im, Kwang Hyuk
    • The Journal of the Korea Contents Association
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    • v.13 no.3
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    • pp.299-306
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    • 2013
  • The purpose of this research is to analyze the questionnaire of oriental medicine and bio-signal data and to identify the correlation between symptoms of oriental medicine and bio-signal in order to develop the customization model for individual patients. First, this research analyzes the questionnaire data using the developed questionnaire for oriental medicine symptoms and selects the optimized questionnaires for identifying the symptoms. Using data mining technologies with the selected questionnaires, this research divides the normal group and the group with symptoms. Finally, this research identifies the correlation between the questionnaire for oriental medicine symptoms and electrogastrogram signal.

Estimation of Effective Moisture Diffusivity of Rapeseed (Brassica napus L.) (유채 종자의 수본확산계수에 관한 연구)

  • Duc, Le Ahn;Hong, Sang-Jin;Han, Jae-Woong;Keum, Dong-Hyuk
    • Journal of Biosystems Engineering
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    • v.33 no.5
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    • pp.296-302
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    • 2008
  • The effective moisture diffusivity and its dependence on drying temperature during drying of rapeseed were experimentally investigated. The data were recorded from thin layer drying experiments at nine different combinations of drying air temperatures of 40, 50, and $60^{\circ}C$ and the relative humidities of 30, 45, and 60%. The moisture diffusion equation was analyzed using stepwise multiple regression analysis. Effective moisture diffusivities were calculated based on the moisture diffusion equation for a spherical shape using Fick's second law. The effective diffusivities during the drying of rapeseed were $l.72{\times}10^{-11}$, $2.41{\times}10^{-11}$ and $3.31{\times}10^{-11}\;m^2{\cdot}s^{-1}$ at 40, 50 and $60^{\circ}C$, respectively. The activation energy for moisture diffusion during drying was $28.47\;kJ{\cdot}mol^{-1}$. The dependence of moisture diffusivity on temperature was described by an Arrhenius-type equation. Drying occurred in the falling rate period and the internal moisture diffusion phenomenon is the governing physical mechanism of the moisture movement in the particles.

Expression of Bacillus macerans Cyclodextrin Glucanotransferase in Bacillus subtilis

  • Kim, Chang-Sup;Han, Nam-Soo;Kweon, Dae-Hyuk;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.230-233
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    • 1999
  • A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for $\alpha$-amylase from B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.

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Highly Time-Resolved Metabolic Reprogramming toward Differential Levels of Phosphate in Chlamydomonas reinhardtii

  • Jang, Cheol-Ho;Lee, Gayeon;Park, Yong-Cheol;Kim, Kyoung Heon;Lee, Do Yup
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1150-1156
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    • 2017
  • Understanding phosphorus metabolism in photosynthetic organisms is important as it is closely associated with enhanced crop productivity and pollution management for natural ecosystems (e.g., algal blooming). Accordingly, we exploited highly time-resolved metabolic responses to different levels of phosphate deprivation in Chlamydomonas reinhardtii, a photosynthetic model organism. We conducted non-targeted primary metabolite profiling using gas-chromatography time-of-flight mass spectrometric analysis. Primarily, we systematically identified main contributors to degree-wise responses corresponding to the levels of phosphate deprivation. Additionally, we systematically characterized the metabolite sets specific to different phosphate conditions and their interactions with culture time. Among them were various types of fatty acids that were most dynamically modulated by the phosphate availability and culture time in addition to phosphorylated compounds.

Prediction of Mechanical Behaviors of Bio-mechanical Materials (생체공학용 척추경 나사의 기계적 거동 예측)

  • Park, Joon-Sik;Choi, Jin Hwa;Cho, Myeong-Woo;Choi, Gil-Woon
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.3 no.1
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    • pp.72-78
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    • 2004
  • In this paper, mechanical behaviors of developed pedicle screw system, made of bio-mechanical materials(Ti-6Al-4V, Grade 5), ale predicted using FEM analysis. As a first step, morphologic construction of normal Korean spines and surgical operation convenience are considered to design optimum pedicle screw system. In this step, various design variables are considered as design parameters to develop optimized models. As a next step, tension and bending tests are performed to improve the structural performance of the developed system using finite element method. In this step, required Static compression and bending test specifications by ASTM F-04 25 04 01 are applied to understand the bio-mechanical behaviors of the designed spinal implant system under various load types. As the results of this research, it is possible to develop efficient pedicle screw system, having enough rigidity and fixation to stand any spinal damage under allowable stress conditions.

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Higher Extraction of Phytochemical Compounds from Tartary Buckwheat Seeds by the Application of Surfactant Formulation

  • Park, Cheol Ho;Azad, Md Obyedul Kalam;Heo, Jeong Won;Kang, Wie Soo
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.686-692
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    • 2017
  • The aim of this study was to determine the suitability of surfactant to extract higher phenolic compound, flavonoid and antioxidant activity from Tartary buckwheat and evaluate the potentiality of surfactant as a screening agent for breeding purpose. Primarily, we employed two types of surfactant (Hydrophilic: Tween 20 and Lipophilic: Span 80) to select the suitable surfactant agent for the extraction of optimum bioactive compounds. Between two surfactants, Tween 20 showed highest efficiency at 4 mM concentration to extract total phenolic content (TP), total flavonoid (TF) and antioxidant activity (AA). Tween 20 at 4 mM concentration was fixed for further analysis along with hot water ($90^{\circ}C$) treatment as a control. In our findings, highest TP (118 mg/g), TF (38 mg/g) and AA (76%) was achieved in KW21 and KW22 among the fifteen accessions of Tartary buckwheat. In other way, TP, TF and AA was 200%, 120% and 110% higher in surfactant formulation compared with control treatment, respectively.

Differential expression of soybean SLTI100 gene encoding translation elongation factor 1A by abiotic stresses

  • Chung, Eun-Sook;Cho, Chang-Woo;So, Hyun-A;Yun, Bo-Hyun;Lee, Jai-Heon
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.255-260
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    • 2009
  • The translation elongation factor 1A, eEF1A, catalyzes the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. By subtractive suppression hybridization technique, we have isolated a soybean low-temperature inducible gene, SLTI100 encoding translation elongation factor 1A. Multiple sequence alignments and phylogenic analysis showed that SLTI100 and other eEF1As originated from diverse organisms are highly conserved. RNA expression of SLTI100 was specifically induced by low temperature, high salt, ABA, or drought stress. Based on the subcellular localization of the corresponding gene product fused to GFP, we were able to confirm that SLTI100-GFP was restricted to the nucleus and cytoplasm. We propose that soybean eEF1A may play an important role in translational regulation during abiotic stress responses in plants.