• Title/Summary/Keyword: Combinatorial library

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Identification of Novel Bioactive Hexapeptides Against Phytopathogenic Bacteria Through Rapid Screening of a Synthetic Combinatorial Library

  • Choi, Jae-Hyuk;Moon, Eun-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.792-802
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    • 2009
  • Antimicrobial peptides (AMPs) are considered to be a promising alternative to conventional antibiotics for future generations. We identified four novel hexapeptides with antimicrobial activity: KCM11 (TWWRWW-$NH_2$), KCM12 (KWRWlW-$NH_2$), KCM21 (KWWWRW-$NH_2$), and KRS22 (WRWFIH-$NH_2$), through positional scanning of a synthetic peptide combinatorial library (PS-SCL). The ability of these peptides to inhibit the growth of a variety of bacteria and unicellular fungi was evaluated. KCM11 and KRS22 preferentially inhibited the normal growth of fungal strains, whereas KCM12 and KCM21 were more active against bacterial strains. Bactericidal activity was addressed in a clear zone assay against phytopathogenic bacteria, including Pectobacterium spp., Xanthomonas spp., Pseudomonas spp., etc. KCM21 showed the highest activity and was effective against a wide range of target organisms. Application of KCM21 with inoculation of Pectobacterium carotovorum subsp. carotovorum on detached cabbage leaves resulted in an immune phenotype or a significant reduction in symptom development, depending on the peptide concentration. Cytotoxicity of the four hexapeptides was evaluated in mouse and human epithelial cell lines using an MTT test. The results revealed a lack of cytotoxic effects.

Identification of Antimicrobial Peptide Hexamers against Oral Pathogens through Rapid Screening of a Synthetic Combinatorial Peptide Library

  • Song, Je-Seon;Cho, Kyung Joo;Kim, Joungmok;Kim, Jeong Hee
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.169-176
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    • 2014
  • A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format $O_{(1-6)}XXXXX-NH_2$, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-$NH_2$ (pep-1) and WXXXXN-$NH_2$ (pep-2), and one positioned peptide, RRRWRN-$NH_2$ (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic ${\alpha}$-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

Combinatorial Synthesis and Screening of the Tb-activated Phosphors in the System CaO-Y2O3-Al2O3 (조합화학을 이용한 CaO-Y2O3-Al2O3계의 Tb활성 형광체의 합성 및 검색)

  • Yoon, Ho-Shin;Kim, Chang-Hae;Kang, Yun-Chan;Ryu, Seung-Kon;Park, Hee-Dong
    • Korean Journal of Materials Research
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    • v.13 no.12
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    • pp.785-790
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    • 2003
  • We have synthesized some phosphors in the system $CaO-Y_2$$O_3$-$Al_2$$O_3$by combinatorial polymerized-complex method. Composition and synthetic temperature of phosphors in the library was screened from the emission intensities of individual samples under VUV excitation. In $Tb^{ 3+}$-activated $CaO-Y_2$$O_3$-$Al_2$$O_3$, green phosphors showing good intensity were found to be X$O_3$$O_{7}$, CaYA1O$_4$, YA1O$_3$, $Y_3$$Al_{5}$$O_{12}$, $Y_4$$A1_2$$O_{9}$ .

Antibody Engineering

  • Hong, Hyo-Jeong;Kim, Sun-Taek
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.3
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    • pp.150-154
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    • 2002
  • Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. An indication of the emerging significance of antibody-based therapeutics is that over a third of the proteins currently undergoing clinical trials in the United States are antibodies. Until the late 1980's, antibody technology relied primarily on animal immunization and the expression of engineered antibodies. However, the development of methods for the expression of antibody fragments in bacteria and powerful techniques for screening combinatorial libraries, together with the accumulating structure-function data base of antibodies, have opened unlimited opportunities for the engineering of antibodies with tailor-made properties for specific applications. Antibodies of low immunogenicity, suitable for human therapy and in vivo diagnosis, can now be developed with relative ease. Here, antibody structure-function and antibody engineering technologies are described.

Screening for bioactive compounds from natural products by ELISA assay

  • Iwanami, Naoko
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.34-37
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    • 1998
  • Combinatorial chemistry is one of the most interested topics in the area of drug discovery. One of the most important points is how to find a lead compound that gives the seed structure for designing of a combinatorial library. Natural products is suitable for searching a new bioactive compound with new structure. We have carried out systematic screening works to find natural products possessing the effects on inter-and intra-cellular signaling. Two hundreds extracts of medical plants and two thousands microbial culture broth samples have been tested for the induction and inhibition of IL-2 or IL-6 production (Fig. 1). ELISA is an efficient method for screenings from such a large number of samples. Now, we apply this method to search prion- binding agents.

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A Novel Screening Strategy for Salt-resistant Alpha-helical Antimicrobial Peptides from a Phage Display Library (Phage Display Library를 이용한 Salt-Resistant Alpha-Helical 항균 펩타이드의 새로운 탐색방법)

  • Park, Ju-Hee;Han, Ok-Kyung;Lee, Baek-Rak;Kim, Jeong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.278-284
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    • 2007
  • A novel screening strategy for salt-resistant antimicrobial peptides from a M13 peptide library was developed. Fusion of MSI-344, a magainin derivative and indolicidin to pIII coat proteins did not significantly affect viability of the recombinant phages, which indicated that the pIII could neutralize toxicity of the antimicrobial peptides and therefore it is possible to construct antimicrobial peptide library in Escherichia coli. On the basis of the conserved sequence of ${\alpha}$-helical antimicrobial peptides, a semi-combinatorial peptide library was constructed in which the peptides were displayed by pIII. To remove hemolytic activity from the library, the phages bound to red blood cells were removed, and the subtracted phage library was screened for binding to target bacteria Pseudomonas aeruginosa and Staphylococcus aureus under high salt concentrations. The screened peptides showed relatively low antimicrobial activity against the target bacteria. However, antimicrobial activities of the screened peptides P06 and S18 were not affected by the cation concentrations of 150 mM $Na^+$, 2 mM $Mg^{2+}$ and 2 mM $Ca^{2+}$ without significant hemolytic activity. This screening strategy that is based on binding capacity to target cells provides new potential to develop salt-tolerant antimicrobial peptides.

Screening of SrO-B2O3-P2O5 Ternary System by Combinatorial Chemistry and QSAR (조합화학과 QSAR를 이용한 SrO-B2O3-P2O5 3원계 청색형광체 개발)

  • Yoo, Jeong-Gon;Back, Jong-Ho;Cho, Sang-Ho;Sohn, Kee-Sun
    • Journal of the Korean Ceramic Society
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    • v.42 no.6 s.277
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    • pp.391-398
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    • 2005
  • It is known that $BaMgAl_{10}O_{17}:Eu^{2+}(BAM)$ phosphors currently used have a serious thermal degradation problem. We screened $SrO-B_2O_3-P_2O_5$ system by a solution combinatorial chemistry technique in an attempt to search for a thermally stable blue phosphor for PDPs. A Quantitative Structure Activity Relationship (QSAR) was also obtained using an artificial neural network trained by the result fiom the combinatorial screening. As a result, we proposed a promising composition range in the $SrO-B_2O_3-P_2O_5$ ternary library. These compositions crystallized into a single major phase, $Sr_6BP_5O_{20}:Eu^{2+}$. The structure of $Sr_6BP_5O_{20}:Eu^{2+}$ was clearly determined by ab initio calculation. The luminescent efficiency of $Sr_6BP_5O_{20}:Eu^{2+}$ was 2.8 times of BAM at Vacuum Ultra Violet (VUV) excitation. The thermal stability was also good but the CIE color chromaticity was slightly poor.

Optimization of VUV Characteristics of M3MgSi2O8:Eu2+ (M=Ca, Sr, Ba) Phosphor by Spray Pyrolysis (분무열분해법을 이용하여 M3MgSi2O8:Eu2+ (M=Ca, Sr, Ba) 형광체 분말의 VUV 특성 최적화)

  • Jung, You-Ri;Jung, Kyeong-Youl
    • Journal of Powder Materials
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    • v.15 no.5
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    • pp.399-404
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    • 2008
  • Spray pyrolysis was applied to prepare $M_{3}MgSi_{2}O_{8}:Eu^{2+}$ (M=Ca, Sr, Ba) blue phosphor powder. The library of a Ca-Sr-Ba ternary system was obtained by a combinatorial method combined with the spray pyrolysis in order to optimize the luminescent property under vacuum ultraviolet (VUV) excitation. 10 potential compositions were chosen from the first screening. The emission shifted to longer wavelength as Ca became a dominant element and the emission intensity was greatly reduced in the composition region at which Ba is dominant element. On the base of the first screening result, the second fine tuning was carried out in order to optimize the luminescence intensity under VUV excitation. The optimal composition for the highest luminescence intensity was $(Ca_{1.7},\;Sr_{0.3},\;Ba_{1.0})Si_{2}O_{8}:Eu^{2+}$ which had the color coordinate of (0.152, 0.072) and about 64% emission intensity of $BaMgAl_{10}O_{17}$ (BAM) phosphor.

Development of New Dihydropyran Linker for Solid-Phase Reaction

  • Ryu, Joon-Hyung;Jeong, Jin-Hyun
    • Archives of Pharmacal Research
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    • v.22 no.6
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    • pp.585-591
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    • 1999
  • The linker which plays a role in connecting a polymer with a scaffold has become an important part n solid-phase reaction. To develop a new linker for alcohols and carbohydrates, dihydropyran moiety was selected in this study. New linker, 1-($4^{l},5^{l}$-dihydro-5H-pyranyl)-7-hydroxyheptan-3-one (5) was synthesized via four steps from $\delta$-valerolactone. This can be called as DDHP-linked Wang resin due to double dihy-dropyran rings. To the one pyran ring of new linker 5 was added Wang resin and other alcohols and carbohydrates as scaffolds were then added successfully to the another pyran ring. Carbohydrate and hydroxyl resins were connected via new linker in a 70% loading yield. The detachment of glucose moiety in the presence of PPTS (2 equiv.) in 1:1 n-buteanol/1,2-dichloroethane at $60^{\circ}C$ for 12 h was carried out quantitatively. When certain combinatorial chemical works are carried out using this dihydropyran linker, Wang resin itself can be recovered. Its fact is particularly very important in industry, because recovered resins can be recycled.

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