• 제목/요약/키워드: Bactericidal activity

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Antibiotic Production of Pseudomonas otitidis PS and Mode of Action (Pseudomonas otitidis PS 균주의 항생물질 생산과 작용 기작)

  • Ahn, Kyung-Joon
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.40-44
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    • 2018
  • An isolate capable of inhibiting the growth of gram-positive bacteria was obtained from the soil of Mushim stream, Cheongju. The isolate was identified as Pseudomonas otitidis PS by 16S rRNA gene sequence analysis. P. otitidis PS produced antibiotics as a secondary metabolite when cultured in 1% soybean meal with 0.5% glucose. The maximum yield was about 0.1%. The antibiotic substance of P. otitidis PS extracted using ethyl acetate displayed a minimum inhibitory concentration of $2{\mu}g/ml$ for Staphylococcus aureus KCTC 1261. The antibiotic substance produced an orange halo on chrome azurol S agar due to siderophore activity. Growth inhibition was decreased when the iron was depleted. Since the antibiotic activity was lost upon the addition of the reducing agent ascorbic acid or during anaerobic culture, it was considered that antibiotic of P. otitidis PS strain exerts its bactericidal effect by the generation of reactive oxygen species.

Screening of Lactobacilli Derived from Fermented Foods and Partial Characterization of Lactobacillus casei OSY-LB6A for Its Antibacterial Activity against Foodborne Pathogens

  • Chung, Hyun-Jung;Yousef, Ahmed E.
    • Preventive Nutrition and Food Science
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    • v.14 no.2
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    • pp.162-167
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    • 2009
  • Various fermented foods were screened in search of food-grade bacteria that produce bacteriocins active against Gram-negative pathogens. An isolate from a mold-ripened cheese presented antibacterial activity against Gram-positive and Gram-negative bacteria. The most active isolate was identified as Lactobacillus casei by a biochemical method, ribotyping, and membrane lipid analysis, and was designated as OSY-LB6A. The cell extracts of the isolate showed inhibition against Escherichia coli p220, E. coli O157, Salmonella enerica serovar Enteritidis, Salmonella Typhimurium, and Listeria monocytogenes. The antibacterial nature of the cell extract from the isolate was confirmed by eliminating the inhibitory effects of acid, hydrogen peroxide, and lytic bacteriophages. The culture supernatant and cell extract retained antibacterial activity after heating at $60{\sim}100^{\circ}C$ for $10{\sim}20$ min. The activity of the cell extract from Lb. casei was eliminated by pronase and lipase. Finally, the cell extract showed a bactericidal mode of action against E. coli in phosphate buffer solution, but it was bacteriostatic in broth medium and food extracts.

Identification and Partial Characterization of Lacticin SA72, a Bacteriocin Produced by Lactococcus lactis SA72 Isolated from Jeot-gal

  • Koo, Kyoung-Mo;Lee, Na-Kyoung;Hwang, Young-Il;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.488-495
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    • 2000
  • Strain SA72 was isolated from Jeot-gal and identified as producer of a bacteriocin, which showed some bactericidal activity against Lactobacillus delbrueckii ATCC 4797. Strain SA72 was tentatively identified as Lactococcus lactis according to the AOI test. Lactococcus lactis SA72 showed a broad spectrum of microorganisms, tested by the modified deferred method. The activity of lacticion SA72, named tentatively as a bacteriocin produced by Lactococcus lactis SA72, was detected during the mid-lon growth phase, reached a maximum during the early stationary phase, and then declined after the late stationary phase. Lacticin SA72 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms when assessed by the spot-on-lawn method. Its anitimicrobial activity on sensitive indicator cells disappeared completely by protease XIV treatment. The inhibitory activity of lacticin SA72 remained after treatment for 15 min at $121^{\circ}C$, 문 was stable in a pH range of 2.0 to 9.0 and all organic solvents examined. It demonstrated a typical bactericidal mode of inhibition against Lactobacillus delbrueckii ATCC 4797. The apparent molecular mass of lacticin SA72 was in the region of 3-3.5 kDa, determined by SDS-PAGE.

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Autophagy in neutrophils

  • Shrestha, Sanjeeb;Lee, Jae Man;Hong, Chang-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.1
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    • pp.1-10
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    • 2020
  • Autophagy is a highly conserved intracellular degradation and energy-recycling mechanism that contributes to the maintenance of cellular homeostasis. Extensive researches over the past decades have defined the role of autophagy innate immune cells. In this review, we describe the current state of knowledge regarding the role of autophagy in neutrophil biology and a picture of molecular mechanism underlying autophagy in neutrophils. Neutrophils are professional phagocytes that comprise the first line of defense against pathogen. Autophagy machineries are highly conserved in neutrophils. Autophagy is not only involved in generalized function of neutrophils such as differentiation in bone marrow but also plays crucial role effector functions of neutrophils such as granule formation, degranulation, neutrophil extracellular traps release, cytokine production, bactericidal activity and controlling inflammation. This review outlines the current understanding of autophagy in neutrophils and provides insight towards identification of novel therapeutics targeting autophagy in neutrophils.

Conserved Virulence Factors of Pseudomonas aeruginosa are Required for Killing Bacillus subtilis

  • Park Shin-Young;Heo Yun-Jeong;Choi Young-Seok;Deziel Eric;Cho You-Hee
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.443-450
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    • 2005
  • The multi-host pathogen, Pseudomonas aeruginosa, possesses an extraordinary versatility which makes it capable of surviving the adverse conditions provided by environmental, host, and, presumably, competing microbial factors in its natural habitats. Here, we investigated the P. aeruginosa-Bacillus subtilis interaction in laboratory conditions and found that some P. aeruginosa strains can outcompete B. subtilis in mixed planktonic cultures. This is accompanied by the loss of B. subtilis viability. The bactericidal activity of P. aeruginosa is measured on B. subtilis plate cultures. The bactericidal activity is attenuated in pqsA, mvfR, lasR, pilB, gacA, dsbA, rpoS, and phnAB mutants. These results suggest that P. aeruginosa utilizes a subset of conserved virulence pathways in order to survive the conditions provided by its bacterial neighbors.

Lysophosphatidylcholine induces azurophil granule translocation via Rho/Rho kinase/F-actin polymerization in human neutrophils

  • Ham, Hwa-Yong;Kang, Shin-Hae;Song, Dong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.3
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    • pp.175-182
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    • 2022
  • Translocation of azurophil granules is pivotal for bactericidal activity of neutrophils, the first-line defense cells against pathogens. Previously, we reported that lysophosphatidylcholine (LPC), an endogenous lipid, enhances bactericidal activity of human neutrophils via increasing translocation of azurophil granules. However, the precise mechanism of LPC-induced azurophil granule translocation was not fully understood. Treatment of neutrophil with LPC significantly increased CD63 (an azurophil granule marker) surface expression. Interestingly, cytochalasin B, an inhibitor of action polymerization, blocked LPC-induced CD63 surface expression. LPC increased F-actin polymerization. LPC-induced CD63 surface expression was inhibited by both a Rho specific inhibitor, Tat-C3 exoenzyme, and a Rho kinase (ROCK) inhibitor, Y27632 which also inhibited LPC-induced F-actin polymerization. LPC induced Rho-GTP activation. NSC23766, a Rac inhibitor, however, did not affect LPC-induced CD63 surface expression. Theses results suggest a novel regulatory mechanism for azurophil granule translocation where LPC induces translocation of azurophil granules via Rho/ROCK/F-actin polymerization pathway.

Production of pediocin by Chemical Synthesis and Bactericidal Mode of Action

  • Koo, Min-Seon;Kim, Wang-June;Kwon, Dea-Young;Min, Kyung-Hee
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.149-153
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    • 2001
  • To investigate the mode of bactericidal action for antimicrobial peptide, pediocin, synthetic and mutant pediocins were prepared by direct chemical synthesis. Native pediocin was purified from Pedio-coccus acidilactici M and its conformational structure and bactericidal functions were analyzed and compared to synthetic pediocin. Schematic mode of pediocin actions, how pediocin binds on the target cell membrane, penetrates and makes tunnel are proposed. For these purposes, primary and secondary structures of pediocin was analyzed and disulfide bond assignment was also done. The pediocin purified from P. acidilactici M had high effective bactericidal ability against gram positive bacteria, especially Listeria monocytogenes and was very stable at extreme pHs and even at high temperatures such as autoclaving temperature (121$^{\circ}C$). Pediocin was consisted of 44 amino acids with four cysteines. Novel synthetic peptides were achieved by solid phase peptide synthesis(SPPS) method. To explain the function of cysteine in C-terminal region, mutant pediocin, Ped[C24A+C44A], was synthesized and their structural and biological functions were analyzed. Second mutant pediocin, Ped[KllE], was prepared to explain the function of lysine at 11 of N-terminal part of pediocin, especially loop of $\beta$-sheet, and to predict the initial binding site of pediocin. The native and synthetic pediocins was showed random coil conformation by spectropolarimetry in moderate conditions. This conformation was observed in extreme conditions such as high temperature and low and high pHs, also. Circular dichroism(CD) data also showed the existence of $\beta$-turn structure in N-terminal part both native and synthetic pediocins. A structural model for pediocin predicts that 18 amino acids in the N-terminal part of the peptide assume a three-strand $\beta$-sheet conformation. This random coil in C-terminal part of pediocin was converted to folding structure, helix structure, in nonpolar solvents such as alcohol and TFE. The disulfide bond between $^{9}$ Cys and $^{14}$ Cys was concrete and inevitable, however, evidences of disulfide bond between $^{24}$ Cys and $^{44}$ Cys was not. Data of Ped[C24A+C44A], pediocin mutant showed that $^{44}$ Cys was required during killing the target cells but not inevitable, since Ped[C24A+C44A] still have bactericidal activity but much less than native pediocin. Another pediocin mutant, Ped[KllE], had still bactericidal activity, was controversial to propose that positive charge like as $^{11}$ Lys in loop or hinge in bacteriocin bound or helped to binding to microorganism with electrostatic interaction between cell membrane especially teichoic acid and positive amino acid nonspecifically. The conformation of pediocin among native, synthetic and mutant pediocins did not show big difference. The conformations between oxidized and reduced pediocin were almost similar regardless of native or synthetic.

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Anti-septic activity of α-cubebenoate isolated from Schisandra chinensis

  • Kook, Minsoo;Lee, Sung Kyun;Kim, Sang Doo;Lee, Ha Young;Hwang, Jae Sam;Choi, Young Whan;Bae, Yoe-Sik
    • BMB Reports
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    • v.48 no.6
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    • pp.336-341
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    • 2015
  • Sepsis is a life-threatening, infectious, systemic inflammatory disease. In this study, we investigated the therapeutic effect of α-cubebenoate, a novel compound isolated from Schisandra chinensis against polymicrobial sepsis in a cecal ligation and puncture (CLP) experimental model. Administration of α-cubebenoate strongly enhanced survival in the CLP model. α-cubebenoate administration also markedly blocked CLP-induced lung inflammation and increased bactericidal activity by enhancing phagocytic activity and hydrogen peroxide generation in mouse bone marrow-derived macrophages and neutrophils. Expression of two important inflammatory cytokines, IL-1 and IL-6, was strongly increased in the CLP model, and this was dramatically blocked by α-cubebenoate. Lymphocyte apoptosis and caspase-3 activation, which are associated with immune paralysis during sepsis, were markedly attenuated by α-cubebenoate. Taken together, our findings indicate that α-cubebenoate, a natural compound isolated from Schisandra chinensis, is a powerful potential anti-septic agent. [BMB Reports 2015; 48(6): 336-341]

Antimicrobial Activity of Continentalic Acid from Aralia cordata Against Enterococcus Strains

  • Jeong, Seung-Il;Yun, Yeon-Hee;Kim, Shin-Moo;Yoon, Kui-Hyun;Kim, Kang-Ju
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.213-216
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    • 2008
  • Continentalic acid (CA, (-)-pimara-8(14), 15-diene-19-oic acid) was isolated from the roots of Aralia cordata (Araliaceae) using bioassay-guided fractionation of a crude chloroform extract. The antibacterial activity of CA against Enterococcus faecalis and Enterococcus gallinarium was estimated by determining minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). CA exhibited potent activity against standard vancomycin-resistant enterococci (VRE) and vancomycin-susceptible enterococci (VSE), with MICs and MBCs values between 4 and $8{\mu}g/mL$ and 4 and $16{\mu}g/mL$, respectively. This compound exhibited potent activity against strains of VRE, which are highly resistant to clinically useful antibiotics. These findings suggest that continentalic acid may be useful in controlling enterococcal infection.

Characterization and Purification of Acidocin 1B, a Bacteriocin Produced by Lactobacillus acidophilus GP1B

  • Han, Kyoung-Sik;Kim, Young-Hoon;Kim, Sae-Hun;Oh, Se-Jong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.774-783
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    • 2007
  • In the present study, acidocin 1B, a bacteriocin produced by Lactobacillus acidophilus GP 1B, exhibited profound inhibitory activity against a variety of LAB and pathogens, including Gram-negative bacteria, and its mode of action was to destabilize the cell wall, thereby resulting in bactericidal lysis. Acidocin 1B was found to be heat stable, because it lost no activity when it was heated up to $95^{\circ}C$ for 60 min. It retained approximately 67% of the initial activity after storage for 30 days at $4^{\circ}C$, and 50% of its initial activity after 30 days at $25^{\circ}C$ and $37^{\circ}C$. The molecular mass of acidocin 1B was estimated to be 4,214.65 Da by mass spectrometry. Plasmid curing results indicated that a plasmid, designated as pLA1B, seemed to be responsible for both acidocin 1B production and host immunity, and that the pLA1B could be transformed into competent cells of L. acidophilus ATCC 43121 by electroporation. Our findings indicate that the acidocin 1B and its producer strain may have potential value as a biopreservative in food systems.