We prepare ZnO nanoparticles by environmentally friendly synthesis using Cyathea nilgiriensis leaf extract. Various phytochemical constituents are identified through the assessment of ethanolic extract of plant Cyathea nilgiriensis holttum by GC-MS analysis. The formation of ZnO nanoparticles is confirmed by FT-IR, XRD, SEM-EDX, TEM, SAED and PSA analysis. TEM observation reveals that the biosynthesized ZnO nanopowder has a hexagonal structure. The calculated average crystallite size from the high intense plane of (1 0 1) is 29.11 nm. The particle size, determined by TEM analysis, is in good agreement with that obtained by XRD analysis. We confirm the formation of biomolecules in plant extract by FT-IR analysis and propose a possible formation mechanism of ZnO nanoparticles. Disc diffusion method is used for the analyses of antimicrobial activity of ZnO nanoparticles. The synthesized ZnO nanoparticles exhibit antimicrobial effect in disc diffusion experiments. The biosynthesized ZnO nanoparticles display good antibacterial performance against B. subtilis (Gram-positive bacteria) and K. pneumonia (Gram-negative bacteria). Bio-synthesized nanoparticles using green method are found to possess good antimicrobial performance.

Polyurethane (PU) nanofibers containing graphene oxide (GO) and Ag doped functionalized reduced graphene oxide (Ag-RGO) were successfully prepared via the electrospinning technique. The uniform distribution of GO sheets along with Ag nanoparticle in the nanofibers was investigated by scanning electron microscopy and the elemental mapping technique. X-ray diffraction and thermal gravimetric analysis verified the presence of GO and Ag in the bicomposite nanofibrous mats. Antibacterial tests against Escherichia coli demonstrated that the addition of GO and Ag-RGO to the PU nanofiber greatly enhanced bactericidal efficiency. Overall, these features of the synthesized nanofibers make them a promising candidate material in the biomedical field for applications such as tissue engineering, wound healing, and drug delivery systems.

Lysozyme was the first humoral antibacterial factor to be studied in insects and it was considered the main immune factor in the haemolymph. Lysozymes are enzymes characterized by their ability to break down bacterial cell walls. We identified four lysozymes(c1, c2, c3, and I type)-encoding cDNAs from P. americana. It was deduced protein sequences are basic in nature, contain 138 amino acids(c1), 139 amino acids(c2), 141 amino acids(c3), and 143 amino acids(I) including conserved cysteine residues. Transcriptional profiles indicated that the predominant form is constitutively expressed and up-regulated upon immune challenge by heat stress. When injected lipoplysaccharide (LPS), lysozymes (I, c1, and c2) was up-regulated in the body. In addition, the expression levels of lysozymes(I, c1, and c2) in the P. americana significantly increased after two hour by injection of LPS. We were cloning and analysis of chitinase from B. germanica. The chitinase was deduced protein sequences are contain 539 amino acid residues long. The chitinase were up-regulated by injection of LPS in B. germanica. the expression levels of chitinase in the B. germanica significantly increased after four hour by injection of LPS. The cDNA encoding the chitinase of B. germanica was expressed as a 67-kDa band in the baculovirus-infected insect cells and the extracts of the recombinant baculovirus-infected cells showed anti-bacterial activity(0.0125㎍/㎖: 2.5±0.3, 0.00125㎍/㎖: 0.6±0.4mm) to C. albican on the plate.

목적: 본 연구는 2-fluoro-4-pyridinecarboxylic acid 및 6-fluoropyridine-3-carboxylic acid를 사용하여 친수성 하이드로젤 콘택트렌즈 재질을 중합하고 제조된 친수성 콘택트렌즈의 물리적, 광학적 특성을 분석하여 이들 물질들의 콘택트렌즈 재료로서의 활용도를 알아보았다. 방법: 본 연구는 카르복시산 및 불소로 치환한 피리딘을 2가지 이상 첨가제로 사용하여 친수성 콘택트렌즈의 기본적인 재질인 HEMA(2-hydroxyethyl methacrylate), AA(acrylic acid), MMA(methylme-thacrylate), 개시제인 AIBN(azobisisobutyronitrile), 그리고 교차결합제인 EGDMA(ethylene glycol dimethacrylate)를 사용하여 공중합하였다. 결과: 생성된 고분자의 물리적 특성을 측정한 결과, 굴절률 1.4348-1.4300, 함수율 33.69~35.84%, 가시광선 투과율 89.6~90.8%, 인장강도 0.2292-0.3344kgf 그리고 접촉각의 경우 76.05°~50.95°의 분포를 나타내었다. 또한 황색포도상구균에 대한 항균실험 결과 2-fluoro-4-pyridinecarboxylic acid 및 6- fluoropyridine-3-carboxylic acid를 5% 첨가했을 때 항균효과가 다소 있는 것으로 나타났다. 결 론: 본 실험 결과, 2-fluoro-4-pyridinecarboxylic acid 및 6-fluoropyridine-3-carboxylic acid를 첨가하여 제조된 친수성 콘택트렌즈는 기본적인 콘택트렌즈 물성을 만족하였으며 우수한 습윤성과 항균성 및 자외선 차단 콘택트렌즈로 활용될 수 있을 것으로 판단된다.

Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of the innate immune system that recognize peptidoglycan, a unique cell wall component of bacteria. Here we cloned and characterized PGRP-S from the bumblebee Bombus ignitus (BiPGRP-S). The BiPGRP-S gene consists of four exons encoding 194 amino acid residues. Comparative analysis indicates that the predicted amino acid sequence of BiPGRP-S shares high identity with enzymatically active PGRP-S proteins and contains the amino acids required for amidase activity. BiPGRP-S in B. ignitus worker bees is constitutively expressed in boththe fat body and epidermis, and it is secreted into the hemolymph. Quantitative real-time PCR assays revealed that in both the fat body and epidermis, the BiPGRP-S gene is highly induced by an injection of Bacillus thuringiensis. In addition, recombinant BiPGRP-S expressed as a 19-kDa protein in baculovirus-infected insect cells can bind to B. megaterium and B. thuringiensis but not to Staphylococcus aureus, Escherichia coli or Beauveria bassiana. Consistent with these data, BiPGRP-S shows antibacterial activity against B. megaterium and B. thuringiensis. These results indicate that BiPGRP-S is an inducible protein that may be involved in the immune response against bacterial infection of the genus Bacillus as an amidase-type PGRP-S.

Attacin is a well-studied glycine-rich antibacterial protein in insect immune response, which has limitary antibacterial effect to some Gram-negative bacteria. A cDNA encoding the attacin gene was screened and isolated from the immunized larvae of the swallowtail butterfly, Papilio xuthus. The complete P. xuthus attacin cDNA is 949 nucleotides encoding a 250 amino acid precursor that contains a putative 18-residue signal peptide, a common 42-residue propeptide sequence and a presumed 190-residue mature protein with a theoretical mass of 19904.01 and a pI of 9.13. The putative mature protein of P. xuthus attacin showed 48%~52% and 24%~30% identity in amino acid sequences with that of lepidopteran and dipteran insects, respectively. The attacin transcript was induced at significant level after injection with bacterial lipopolysaccharide (LPS). Recombinant attacin was highly expressed in E. coli BL21 (DE3) cells by fusing with an N-terminal S-tag/thrombin cleavage site configuration protein to avoid the cell death during induction. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC). After desalting and cleavage with thrombin, the recombinant attacin was released and showed considerably antibacterial activity against Gram-negative bacteria, E. coli ML 35. Our results proved that this protein family with a potent antibacterial activity may play a role in the immune response of butterflies.

To find some antibacterial peptides responsible for bacterial resistance, we performed differential hybridization with total cDNA probes which synthesized from normal and immunized larvae. Thirteen individual cDNA transcripts were expressed differentially in a total 1,862 random cDNA clones. One of upregulated genes is a novel member of the insect defensin-like peptide(Coprisin), a family of antibacterial peptide. Northern blot analysis showed that Coprisin was up-regulated at 4h and reached the highest point level at 16h after injection of E.coli. The deduced amino acid sequence of Coprisin was composed of 80 amino acids with predicted molecular weight of 8.6 kDa and PI of 8.72. Comparison of the deduced amino acid mature portion of Coprisin with defensin-like peptide of other insect indicated that it has 79.1% and 67.4% identity with Anomala cuprea and Allomyrina dichotoma, respectively. To find antibacterial active region of Coprisin, we synthesized four peptides corresponding to amino acid residues 1V-43N-NH2(CopN1), 5-16(CopN2), 19-30(CopN3) and 31-43(CopN4) of coprisin having amidated amino acid residues at their Cterminal. A 12-mer amidated at its C-terminus, ACALHCIALRKK-NH2 (Ala19-Lys30-NH2) was synthesized based on the deduced amino acid sequence, assumed to be an active site sequence. This peptides showed antibacterial activity against E.coli, Staphylococcus aureus, MRSA, Psedomonas syringae, and Pectobacterium carotovorium. Modified 9-mer peptide, LRCIALRKK-NH2, showed strong antibacterial activity than mellitin peptide used as a positive control against gram-negative and gram-positive bacteria. This peptide showed no haemolytic activity and quite stable at 100℃ for several hours of incubation and in a wide pH range(pH2-12). Therefore, this peptide may be a good candidate for the development of new drug with potent antibacterial activity without cytotoxicity.

Drug delivery system(DDS) have been actively studied for the past twenty years. Dual action agents are unique chemical entities comprised of two different types of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. In spite of the advent of the antibacterial agent the sulfa agents are the most widely used antibacterial agent today. In this study, new antibacterials derivative was synthesized using glutaraldehyde such as crosslinking agent for the purpose of dual-action as DDS study. Antibacterial activity of these new synthetic derivative between their structures and activities were examined by disc diffusion method. As a result, new synthetic derivative exhibited the broad antibacterial activities against Gram(+) and Gram(-) bacilli. Especially, the antibacterial effect of new synthetic derivative against Gram negative(Esherichia. coli) was much stronger than that against Gram positive.

대두 발효식품으로부터 분리 동정한 B. subtilis HS 25가 생산한 항균물질의 특성을 검토하였다. 열안정성을 검토한 결과, 에서 10분 처리에서는 항균활성이 전혀 떨어지지 않았지만 15분 이상의 처리에서는 항균활성이 약간 떨어지는 경향을 보였다. pH 안정성은 pH 의 산성측보다 pH 까지의 알칼리 범위에서 안정된 항균활성을 유지하였다. 특히, E. coli 및 P. mirabilis에 대한 항균활성은 pH 12.0에서도 대조구와 거의 대

전통발효식품의 기능성 및 저장성을 증진시킬 목적으로 전통된장으로부터 효소활성 및 항균활성이 우수한 CH-14 균주를 분리하여 그 특성을 검토하였다. 분리한 CH-14 균주는 그람양성, catalase 양성, oxidase 음성, 내생포자 및 편모를 가지는 간균으로 세포의 크기는 이었다. 최종 분리균을 Bergey's Manual of Systematic Bacteriology 및 Bergey's Manual of Determinative Bact