• Title/Summary/Keyword: Representative Strain

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The Overexpression of Subtilisin Enzyme Using Mutations on Transition State Regulatory Proteins of AprE Promoter and Development of Bacillus subtilis Host System (AprE Promoter전이상태 조절인자 변이주를 이용한 공업적 효소의 과발현과 고초균 숙주계의 개발)

  • 류성호;박승환김병기
    • KSBB Journal
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    • v.11 no.1
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    • pp.8-14
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    • 1996
  • Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

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Two-dimensional gel Electrophoresis of Helicobacter pylori for Proteomic Analysis

  • Jung, Tae-Sung;Kang, Seung-Chul;Choi, Yeo-Jeong;Jeon, Beong-Sam;Park, Jeong-Won;Jung, Sun-Ae;Song, Jae-Young;Choi, Sang-Haeng;Park, Seong-Gyu;Choe, Mi-Young;Lee, Byung-Sang;Byun, Eun-Young;Baik, Seung-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.97-108
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    • 2000
  • Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.

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Continuum Based Plasticity Models for Cubic Symmetry Lattice Materials Under Multi-Surface Loading (다중면 하중하에 정방향 대층구조를 가진 격자재료의 연속적인 소성모델)

  • Seon, Woo-Hyun;Hu, Jong-Wan
    • Journal of the Korean Society for Advanced Composite Structures
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    • v.2 no.3
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    • pp.1-11
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    • 2011
  • The typical truss-lattice material successively packed by repeated cubic symmetric unit cells consists of sub-elements (SE) proposed in this study. The representative continuum model for this truss-lattice material such as the effective strain and stress relationship can be formulated by the homogenization procedure based on the notation of averaged mechanical properties. The volume fractions of micro-scale struts have a significant influence on the effective strength as well as the relative density in the lattice plate with replicable unit cell structures. Most of the strength contribution in the lattice material is induced by axial stiffness under uniform stretching or compression responses. Therefore, continuum based constitutive models composed of homogenized member stiffness include these mechanical characteristics with respect to strength, internal stress state, material density based on the volume fraction and even failure modes. It can be also recognized that the stress state of micro-scale struts is directly associated with the continuum constitutive model. The plastic flow at the micro-scale stress can extend the envelope of the analytical stress function on the surface of macro-scale stress derived from homogenized constitutive equations. The main focus of this study is to investigate the basic topology of unit cell structures with the cubic symmetric system and to formulate the plastic models to predict pressure dependent macro-scale stress surface functions.

Efficient Adaptive Finite Element Mesh Generation for Dynamics (동적 문제에 효율적인 적응적 유한요소망)

  • Yoon, Chongyul
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.26 no.5
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    • pp.385-392
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    • 2013
  • The finite element method has become the most widely used method of structural analysis and recently, the method has often been applied to complex dynamic and nonlinear structural analyses problems. Even for these complex problems, where the responses are hard to predict, finite element analyses yield reliable results if appropriate element types and meshes are used. However, the dynamic and nonlinear behaviors of a structure often include large deformations in various portions of the structure and if the same mesh is used throughout the analysis, some elements may deform to shapes beyond the reliable limits; thus dynamically adapting finite element meshes are needed in order for the finite element analyses to be accurate. In addition, to satisfy the users requirement of quick real run time of finite element programs, the algorithms must be computationally efficient. This paper presents an adaptive finite element mesh generation scheme for dynamic analyses of structures that may adapt at each time step. Representative strain values are used for error estimates and combinations of the h-method(node movement) and the r-method(element division) are used for mesh refinements. A coefficient that depends on the shape of an element is used to limit overly distorted elements. A simple frame example shows the accuracy and computational efficiency of the scheme. The aim of the study is to outline the adaptive scheme and to demonstrate the potential use in general finite element analyses of dynamic and nonlinear structural problems commonly encountered.

Transcriptional Analysis Responding to Propanol Stress in Escherichia coli (대장균에서 프로판올 스트레스에 관한 전사분석)

  • Park, Hye-Jin;Lee, Jin-Ho
    • Journal of Life Science
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    • v.22 no.3
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    • pp.417-427
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    • 2012
  • We compared the transcriptome in response to propanol stress in wild-type and propanol-resistant mutant Escherichia coli using the DNA microarray technique. The correlation value of RNA expression between the propanol-treated wild type and the untreated-one was about 0.949, and 50 genes were differentially expressed by more than twofold in both samples. The correlation value of RNA expression between the propanol-treated mutant and the untreated one was about 0.951, and 71 genes in two samples showed differential expression patterns. However, the values between the wild type and mutant, regardless of propanol addition, were 0.974-0.992 and only 1-2 genes were differentially expressed in the two strains. The representative characteristics among differentially expressed genes in W3110 or P19 treated with propanol compared to untreated samples were up-regulation of hest shock response genes and down-regulation of genes relating to ribosome biosynthesis. In addition, many genes were regulated by transcription regulation factors such as ArcA, CRP, FNR, H-NS, GatR, or PurR and overexpressed by sigma factor RpoH. We confirmed that RpoH mediated an important host defense function in propanol stress in E. coli W3110 and P19 by comparison of cell growth rate among the wild type, rpoH disruptant mutant, and rpoH-complemented strain.

Proteolytic Yeasts Isolated from Mackerel (Scomber japonicus) (고등어에서 분리된 부패성 효모)

  • OH Eun-Gyong;PARK Mi-Yeon;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.471-476
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    • 1998
  • Microbiological spoilage of marine fish is complex process occurring by bacteria, yeasts and molds. There have been rare study for saprophytic yeasts although having enormous numbers of bacteriological studies on the spoilage of marine fish. The 14 genera of yeasts isolated from mackerel (Scomber japonicus) with high frequency of occurrence were Candida sp., Rhodotorula sp., Torulopsis sp., Cryptotoccus sp. and Tricosporon sp. Among these ones Candida lipolytica was identified as the strongest proteolytic yeast, then named Candida lipolytica FM5 (C. lipolytica FM5). C. lipolytica FM5 showed optimum growth at $25^{\circ}C$, pH 7.0 and could grow at $5^{\circ}C$ and in medium containing $10\%$ sodium chloride, To evaluate the saprophytic activity of the selected strain, C, lipolytica FM5 and Pseudomonas fluorescens ATCC 17571 which is one of representative spoilage bacteria were individually inoculated into the sterilized fish muscle homogenates, and then pH changes and volatile basic nitrogen (VBN) values were checked during the storage at various temperatures. According to the experimental results, the productions of VBN by C. lipolytica FM5 in the fish muscle homogenates were 50 mg-N/100 g at $5^{\circ}C$, 152 mg-N/100 g at $15^{\circ}C$ and 379 mg-N/100 g at $25^{\circ}C$ for 1 week storage, respectively. Above results were nearly same as in case of Ps. fluorescens ATCC 17571 inoculation. It suggest that sapyophytic yeasts also have important role in spoilage of marine fish.

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Deduction of Correlations between Shear Wave Velocity and Geotechnical In-situ Penetration Test Data (전단파속도와 지반공학적 현장 관입시험 자료의 상관관계 도출)

  • Sun, Chang-Guk;Kim, Hong-Jong;Chung, Choong-Ki
    • Journal of the Earthquake Engineering Society of Korea
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    • v.12 no.4
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    • pp.1-10
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    • 2008
  • Shear wave velocity($V_S$), which can be obtained using various seismic tests, has been emphasized as representative geotechnical dynamic characteristic mainly for seismic design and seismic performance evaluation in the engineering field. For the application of conventional geotechnical site investigation techniques to geotechnical earthquake engineering, standard penetration tests(SPT) and piezocone penetration tests(CPTu) together with a variety of borehole seismic tests were performed at many sites in Korea. Through statistical modeling of the in-situ testing data, in this study, the correlations between $V_S$ and geotechnical in-situ penetrating data such as blow counts(N value) from SPT and piezocone penetrating data such as tip resistance ($q_t$), sleevefriction($f_s$), and pore pressure ratio($B_q$) were deduced and were suggested as an empirical method to determine $V_S$. Despite the incompatible strain levels of the conventional geotechnical penetration tests and the borehole seismic tests, it is shown that the suggested correlations in this study are applicable to the preliminary estimation of $V_S$ for Korean soil layers.

Lactobacillus plantarum APsulloc 331261 Fermented Products as Potential Skin Microbial Modulation Cosmetic Ingredients (Lactobacillus plantarum APsulloc 331261 발효 용해물의 피부 미생물 조절 효과)

  • Kim, Hanbyul;Myoung, Kilsun;Lee, Hyun Gee;Choi, Eun-Jeong;Park, Taehun;An, Susun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.1
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    • pp.23-29
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    • 2020
  • The skin is colonized by a large number of microorganisms with a stable composition of species. However, disease states of skin such as acne vulgaris, psoriasis, and atopic dermatitis have specific microbiome compositions that are different from those of healthy skin. The target modulation of the skin microbiome can be a potential treatment for these skin diseases. Quorum sensing (QS), a bacterial cell-cell communication system, can control the survival of bacteria and increase cell density. Also, QS affects the pathogenicity of bacteria such as biofilm formation and protease production. In this study, we confirmed anti-QS activity of Amorepacific patented ingredients, which are Lactobacillus ferment lysate (using Lactobacillus plantarum APsulloc 331261, KCCM 11179P) through bio-reporter bacterial strain Chromobacterium violaceum. The purple pigment production of C. violaceum controlled by QS was reduced 27.3% by adding 10 ㎍/mL of Lactobacillus ferment lysate (freeze dried). In addition, the Lactobacillus ferment lysate increased growth of Staphylococcus epidermidis 12% and decreased growth of Pseudomonas aeruginosa 38.5% and its biofilm formation 17.7% at a concentration of 10 ㎍/mL compared to the untreated control group. Moreover, S. epidermidis was co-cultured with the representative dermatological bacterium Staphylococcus aureus in the same genus, the growth of S. epidermidis was increased 134 % and the growth of S. aureus was decreased 13%. These results suggest that fermented lysate using Lactobacillus plantarum APsulloc 331261 may be useful as a cosmetic ingredient that can control the balance of skin microbiome.

Effect of 850 nm near-infrared light emitting diode irradiation on the production of 5-aminolevulinic acid in Rhodobacter sphaeroides (Rhodobacter sphaeroides에서 5-aminolevulinic acid 생산에 대한 850 nm 근적외선 발광다이오드 조사 효과)

  • Mo, SangJoon
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.217-223
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    • 2021
  • 5-aminolevulinic acid (ALA) is a representative photosensitizer used in numerous fields including cancer diagnosis and treatment. In this study, experiments were conducted to optimize the growth of Rhodobacter sphaeroides and production of ALA through LED irradiation of various wavelengths, addition of organic acid precursors of ALA, and changes in glucose concentration. After 72 h cultivation, the 850 nm wavelength LED irradiated at the same light intensity as the incandescent lamp increased the growth of R. sphaeroides and the production of ALA about 1.5- and 1.8-fold as compared with the control, respectively (p <0.0001 and p <0.0001). As a result of culturing R. sphaeroides by irradiating an LED with a wavelength of 850 nm after adding organic acid to the final concentration of 5 mM in culture medium, the production of ALA was increased about 2.8-fold in medium supplemented with pyruvic acid compared with the control (p <0.0001). In addition, the growth of the strain and the production of ALA were increased about 2.9- and 3.4-fold in medium supplemented with 40 mM glucose compared to the control which added only 5 mM pyruvic acid, respectively (p <0.0001 and p <0.0001). The yield of ALA per cell dry mass was about 1.4 folds higher than that of the control in 20 and 40 mM glucose, respectively (p <0.001). In conclusion, the growth of R. sphaeroides and production of ALA were increased by 850 nm wavelength LED irradiation. It also optimized the growth of R. sphaeroides and production of ALA through organic acid addition and glucose concentration changes.

Characterization of broad bean wilt virus 2 isolated from Perilla frutescens in Korea (국내 잎들깨에서 발생한 잠두위조바이러스2의 특성 구명)

  • Hyun-Sun Kim;Hee-Seong Byun;You-Ji Choi;Hyun-Yong Choi;Jang-Kyun Seo;Hong-Soo Choi;Bong-Choon Lee;Mikyeong Kim;Hae-Ryun Kwak
    • Korean Journal of Environmental Biology
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    • v.41 no.1
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    • pp.1-13
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    • 2023
  • Broad bean wilt virus 2 (BBWV2) is a species in the genus Fabavirus and family Secoviridae, which is transmitted by aphids and has a wide host range. The BBWV2 genome is composed of two single-stranded, positive-sense RNAs, RNA-1 and RNA-2. The representative symptoms of BBWV2 are mosaic, mottle, vein clearing, wilt, and stunting on leaves, and these symptoms cause economic damage to various crops. In 2019, Perilla fructescens leaves with mosaic and yellowing symptoms were found in Geumsan, South Korea. Reverse-transcription polymerase chain reaction (RT-PCR) was performed with specific primers for 10 reported viruses, including BBWV2, to identify the causal virus, and the results were positive for BBWV2. To characterize a BBWV2 isolate (BBWV2-GS-PF) from symptomatic P. fructescens, genetic analysis and pathogenicity tests were performed. The complete genomic sequences of RNA-1 and RNA-2 of BBWV2-GS-PF were phylogenetically distant to the previously reported BBWV2 isolates, with relatively low nucleotide sequence similarities of 76-80%. In the pathogenicity test, unlike most BBWV2 isolates with mild mosaic or mosaic symptoms in peppers, the BBWV2-GS-PF isolate showed typical ring spot symptoms. Considering these results, the BBWV2-GS-PF isolate from P. fructescens could be classified as a new strain of BBWV2.