• Title/Summary/Keyword: surface-displayed

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Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets

  • Kim, Juhyung;Kim, Hyung Kyu;Jang, Hye Jeong;Kim, Eunkyung;Kim, Moon Kyu
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.547-553
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    • 2015
  • The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.

Comparison of the Organophosphorus Hydrolase Surface Display Using InaVN and Lpp-OmpA Systems in Escherichia coli

  • Karami, Ali;Latifi, Ali Mohamad;Khodi, Samaneh
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.379-385
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    • 2014
  • The purpose of this study was to compare the ability of an engineered Escherichia coli to degrade chlorpyrifos (Cp) using an organophosphorus hydrolase enzyme, encoded in both Flavobacterium sp. ATCC 27551 or Pseudomonas diminuta, by employing the Lpp-OmpA chimera and the N-terminal domain of the ice nucleation protein as anchoring motifs. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by both anchors on the outer membrane. This is the first report on the presentation of OPH on the cell surface by Lpp-OmpA under the control of the T7 promoter. The results showed cell growth in the presence of Cp as the sole source of energy, without growth inhibition, and with higher whole-cell activity for both cells harboring plasmids pENVO and pELMO, at approximately 10,342.85 and 10,857.14 U/mg, respectively. Noticeably, the protein displayed by pELMO was lower than the protein displayed by pENVO. It can be concluded that Lpp-OmpA can display less protein, but more functional OPH protein. These results highlight the high potential, of both engineered bacteria, for use in the bioremediation of pesticide-contaminated sources in the environment.

Effect of Al and Cr contents on the High Temperature Oxidation- and Sulfidation-resistance of Fe Alloys (Fe합금의 내 산화성과 황화성에 미치는 Al과 Cr 함량의 영향)

  • Kim, Seul-Ki;Lee, Dong-Bok
    • Journal of the Korean institute of surface engineering
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    • v.45 no.2
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    • pp.61-69
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    • 2012
  • Alloys of Fe-(5, 10, 15)Al and Fe-(10, 20, 30, 40)Cr were corroded at 700 and $800^{\circ}C$ for 70 hr in either atmospheric air or 1 atm of Ar+$1%SO_2$ gases. In these atmospheres, Fe-5Al and Fe-10Cr alloys displayed poor corrosion resistance. In atmospheric air, Fe-5Al alloys formed oxide nodules, while Fe-10Cr alloys formed thick scales and internal oxides. In Ar+$1%SO_2$ gases, Fe-5Al and Fe-10Cr alloys formed thick, nonadherent bi-layered scales, which grew primarily by the outward diffusion of Fe ions and inward diffusion of oxygen and sulfur ions. By contrast, in atmospheric air and Ar+$1%SO_2$ gases, Fe-(10, 15)Al and Fe-(20, 30, 40)Cr alloys displayed good corrosion resistance by forming $Al_2O_3$ and $Cr_2O_3$ layers on the surface, respectively.

Characterization of Agarose Produced by Yeast Cell Surface Displayed-Arylsulfatase (효모 표층 Arylsulfatase에 의해 제조된 Agarose의 특성)

  • Cho, Eun-Soo;Kim, Jeong-Hwan;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.428-433
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    • 2010
  • Enzymatic hydrolysis of sulfate groups in agaropectin or agar simplifies the production process of high-quality or low sulfate-content agarose. This study was investigated that cell surface displayed arylsulfatase can be applied to desulfatation of agar for production of agarose. Sulfate content of agarose prepared by treatment of yeast surface-displayed arylsulfatase was decreased in a enzyme dosedependent manner. Especially, 35 unit/mL of yeast surface arylsulfatase attenuated sulfate content of agarose up to 0.2%. In the 0.6% agar(Junsei), 35 unit/mL enzyme treated at $40^{\circ}C$ for 3 h showed the lowest content of sulfate. Therefore, this result was determined to be the optimal condition to desulfatation of agar for production of agarose. In addition, the gel strength of yeast surface arylsulfatase treated agar and commercial agarose were compared. Agarose prepared by treatment of yeast surface arylsulfatase showed $559.8{\pm}0.12$ of gel strength, and it is a similar compared to the commercial agarose.

Removal of Chromium (VI) by Escherichia coli Cells Expressing Cytoplasmic or Surface-Displayed ChrB: a Comparative Study

  • Zhou, Xiaofeng;Li, Jianghui;Wang, Weilong;Yang, Fan;Fan, Bingqian;Zhang, Chenlu;Ren, Xiaojun;Liang, Feng;Cheng, Rong;Jiang, Fengying;Zhou, Huaibin;Yang, Juanjuan;Tan, Guoqiang;Lyu, Jianxin;Wang, Wu
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.996-1004
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    • 2020
  • Various genetically engineered microorganisms have been developed for the removal of heavy metal contaminants. Metal biosorption by whole-cell biosorbents can be enhanced by overproduction of metal-binding proteins/peptides in the cytoplasm or on the cell surface. However, few studies have compared the biosorption capacity of whole cells expressing intracellular or surface-displayed metal-adsorbing proteins. In this study, several constructs were prepared for expressing intracellular and surface-displayed Ochrobactrum tritici 5bvl1 ChrB in Escherichia coli BL21(DE3) cells. E. coli cells expressing surface-displayed ChrB removed more Cr(VI) from aqueous solutions than cells with cytoplasmic ChrB under the same conditions. However, intracellular ChrB was less susceptible to variation in extracellular conditions (pH and ionic strength), and more effectively removed Cr(VI) from industrial wastewater than the surface-displayed ChrB at low pH (<3). An adsorption-desorption experiment demonstrated that compared with intracellular accumulation, cell-surface adsorption is reversible, which allows easy desorption of the adsorbed metal ions and regeneration of the bioadsorbent. In addition, an intrinsic ChrB protein fluorescence assay suggested that pH and salinity may influence the Cr(VI) adsorption capacity of ChrB-expressing E. coli cells by modulating the ChrB protein conformation. Although the characteristics of ChrB may not be universal for all metal-binding proteins, our study provides new insights into different engineering strategies for whole-cell biosorbents for removing heavy metals from industrial effluents.

A Study on the Surface Roughness in Grinding Process by Frequency Analysis (평면연삭(平面硏削)에서 주파수(周波數) 분석(分析)에 의한 표면(表面) 거칠기 해석(解釋))

  • Kim, G.N.;Cheong, C.C.;Kim, K.S.
    • Journal of the Korean Society for Nondestructive Testing
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    • v.11 no.2
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    • pp.32-41
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    • 1991
  • This paper is to analyze the surface roughness in wet and dry surface grinding in which working conditions are table feed, cross feed and down feed. FFT analyzer is used to identify the surface roughness and the data of surface roughness are stored and displayed using an interfaced computer. In results, components affecting surface roughness exist within 200Hz with respect the conditions and methods of grinding operation. Also, with increase of table feed, cross feed and down feed, the value of surface roughness increases. Of them, cross feed has first influence on surface roughness. In dry, preciser surface can be obtained than one in wet.

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Corrosion and Surface Resistance of Ni-C Composite by Electrodeposition (전해도금에 의한 Ni-C 복합층의 내식성 및 표면 전기저항)

  • Park, Je-Sik;Lee, Sung-Hyung;Jeong, Goo-Jin;Lee, Churl-Kyoung
    • Korean Journal of Materials Research
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    • v.21 no.5
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    • pp.288-294
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    • 2011
  • Simultaneous Ni and C codeposition by electrolysis was investigated with the aim of obtaining better corrosion resistivity and surface conductivity of a metallic bipolar plate for application in fuel cells and redox flow batteries. The carbon content in the Ni-C composite plate fell in a range of 9.2~26.2 at.% as the amount of carbon in the Ni Watt bath and the roughness of the composite were increased. The Ni-C composite with more than 21.6 at.% C content did not show uniformly dispersed carbon. It also displayed micro-sized defects such as cracks and crevices, which result in pitting or crevice corrosion. The corrosion resistance of the Ni-C composite in sulfuric acid is similar with that of pure Ni. Electrochemical test results such as passivation were not satisfactory; however, the Ni-C composite still displayed less than $10^{-4}$ $A/cm^2$ passivation current density. Passivation by an anodizing technique could yield better corrosion resistance in the Ni-C composite, approaching that of pure Ni plating. Surface resistivity of pure Ni after passivation was increased by about 8% compared to pure Ni. On the other hand, the surface resistivity of the Ni-C composite with 13 at.% C content was increased by only 1%. It can be confirmed that the metal plate electrodeposited Ni-C composite can be applied as a bipolar plate for fuel cells and redox flow batteries.

A Study of Geometric Modeling for Ship Hull Forms Using Open Uniform B-spline Surface (Open 균일 B-spline 곡면을 이용한 선체 곡면 표현에 관한 연구)

  • H.K. Shin;K.W. Park
    • Journal of the Society of Naval Architects of Korea
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    • v.28 no.2
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    • pp.21-27
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    • 1991
  • This paper outlines the method of formulating the bi-cubic B-spline surface of ship hull, employing the open uniform knot vector as well as the periodic uniform knot vector. An appropriate set of B-spline control vertices to generate the B-spline surface is determined by obtaining the pseudoinverse matrix of basis functions. The comparison between the given offsets and the resulting coordinates from the generated ship hull surface shows a good agreement. To check the fairness of the surface Gaussian curvature is calculated on many small subpatches and displayed on the black-and-white plot of the isoparametric net of the surface.

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