• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.759-764
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• 2015

Antimicrobial peptides (AMPs) are one of the critical components in host innate immune responses to imbalanced and invading microbial pathogens. Although the antimicrobial activity and mechanism of action have been thoroughly investigated for decades, the exact biological properties of AMPs are still elusive. Most AMPs generally exert the antimicrobial effect by targeting the microbial membrane, such as barrel stave, toroidal, and carpet mechanisms. Thus, the mode of action in model membranes and the discrimination of AMPs to discrepant lipid compositions between mammalian cells and microbial pathogens (cell selectivity) have been studied intensively. However, the latest reports suggest that not only AMPs recently isolated but also well-known membrane-disruptive AMPs play a role in intracellular killing, such as apoptosis induction. In this mini-review, we will review some representative AMPs and their antimicrobial mechanisms and provide new insights into the dual mechanism of AMPs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.5
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• pp.745-751
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• 2003

This study was carried out to investigate whether peptide produced from wheat protein by enzyme hydrolysis can be used as a natural antimicrobial agent. Antimicrobial peptide was obtained from wheat protein hydrolyzed by 7 of pretense. The produced antimicrobial peptide was purified through ultrafiltration, membrane filtration and HPLC and molecular weight and amino acid sequence of the purified antimicrobial peptide were determined. Among hydrolysate produced from wheat protein by 7 of protease, antimicrobial activity was observed for the peptide obtained from Asp. saito protease. The Asp. saito protease did produce antimicrobial hydrolysate showing the highest antimicrobial activity at reaction condition of 37$^{\circ}C$ and pH 6.0, but not at reaction condition above 5$0^{\circ}C$. Wheat protein hydrolysate was fractionated by membrane filtration and showed antimicrobial activity between molecular weight 1,000~3,000. The antimicrobial activity fraction obtained by membrane filtration was separated through HPLC and showed antimicrobial activity in the peak of retention time 31.1~31.8 min. We could convince this hydrolysate as heat-stable peptide since antimicrobial activity was maintained after treated with heat for 15 min at 121$^{\circ}C$. Molecular weight of antimicrobial peptide identified by MALDI-mass was 1,633. Amino acid sequence of antimicrobial peptide was cysteine, glycine, prolin, prolin, prolin, valine, valine, alanine, alanine and arginine.

• Korean Journal of Agricultural Science
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• v.29 no.2
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• pp.78-90
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• 2002

This study was carried out to investigate whether peptide produced from wheat protein by enzyme hydrolysis can be used as a natural antimicrobial agent. Antimicrobial peptide was obtained from wheat protein by protease of 7 species. The produced antimicrobial peptide was purified through ultrafiltration, membrane filtration and HPLC, and molecular weight and amino acid sequence of the purified antimicrobial peptide were determined. Among hydrolysate produced from wheat protein by protease of 7 species, antimicrobial activity was observed for the peptide obtained from Asp. saito protease. The Asp. saito protease did production antimicrobial hydrolysate showing the highest antimicrobial activity at reaction condition of $37^{\circ}C$ and pH 6.0, but not at reaction condition above $50^{\circ}C$. Wheat protein hydrolysate was fractionated by membrane filtration and showed antimicrobial activity between molecular weight 1,000 - 3,000. The antimicrobial activity fraction obtained by membrane filtration was separated through HPLC and showed antimicrobial activity in the peak of retention time 31.1 - 31.8 min. Since after wheat protein protease hydrolysate was heated during 15 min at $121^{\circ}C$, antimicrobial activity was maintained, we could be conviction as heat-stable peptide. Molecular weight of antimicrobial peptide identified by MALDI-mass was 1,633. Amino acid sequence of antimicrobial peptide was cysteine, glycine, prolin, prolin, prolin, valine, valine, alanine, alanine and arginine.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.1-10
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• 2019

Gram-negative pathogens, such as Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii, pose a serious threat to public health worldwide, due to high rates of antibiotic resistance and the lack of development of novel antimicrobial agents targeting Gram-negative bacteria. The outer membrane (OM) of Gram-negative bacteria is a unique architecture that acts as a potent permeability barrier against toxic molecules, such as antibiotics. The OM is composed of phospholipids, lipopolysaccharide (LPS), outer membrane ${\beta}-barrel$ proteins (OMP), and lipoproteins. These components are synthesized in the cytoplasm or in the inner membrane, and are then selectively transported to the OM by the specific transport machines, including the Lol, BAM, and Lpt pathways. In this review, we summarize recent studies on the assembly systems of OM components and analyze studies for the development of inhibitors that target these systems. These analyses show that OM assembly machines have the potential to be a novel attractive drug target of Gram-negative bacteria.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.245-253
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• 2014

Antimicrobial peptides (AMPs) are small but effective cationic peptides with variable length. In previous study, four hexapeptides were identified that showed antimicrobial activities against various phytopathogenic bacteria. KCM21, the most effective antimicrobial peptide, was selected for further analysis to understand its modes of action by monitoring inhibitory effects of various cations, time-dependent antimicrobial kinetics, and observing cell disruption by electron microscopy. The effects of KCM21 on Gram-negative strain, Pseudomonas syringae pv. tomato DC3000 and Gram-positive strain, Clavibacter michiganensis subsp. michiganensis were compared. Treatment with divalent cations such as $Ca^{2+}$ and $Mg^{2+}$ inhibited the bactericidal activities of KCM21 significantly against P. syringae pv. tomato DC3000. The bactericidal kinetic study showed that KCM21 killed both bacteria rapidly and the process was faster against C. michiganensis subsp. michiganensis. The electron microscopic analysis revealed that KCM21 induced the formation of micelles and blebs on the surface of P. syringae pv. tomato DC3000 cells, while it caused cell rupture against C. michiganensis subsp. michiganensis cells. The outer membrane alteration and higher sensitivity to $Ca^{2+}$ suggest that KCM21 interact with the outer membrane of P. syringae pv. tomato DC3000 cells during the process of killing, but not with C. michiganensis subsp. michiganensis cells that lack outer membrane. Considering that both strains had similar sensitivity to KCM21 in LB medium, outer membrane could not be the main target of KCM21, instead common compartments such as cytoplasmic membrane or internal macromolecules might be a possible target(s) of KCM21.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.429-438
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• 2020

The emergence of multi-drug resistant, pathogenic methicillin-resistant Staphylococcus aureus (MRSA) is a threat to global health and has created a need for novel functional therapeutic agents. In this study, we evaluated the underlying mechanisms of the anti-MRSA effect of an azaphilone pigment, sclerotiorin (SCL) from Penicillium sclerotiorum. The antimicrobial activity of SCL was evaluated using agar disc diffusion, broth microdilution, time-kill assays and biophysical studies. SCL exhibits selective activity against Gram positive bacteria including MRSA (range, MIC = 128-1028 ㎍/ml) and exhibited rapid bactericidal action against MRSA with a > 4 log reduction in colony forming units within three hours of administration. Biophysical studies, using fluorescent probes and laser or electron microscopy, demonstrated a SCL dose-dependent alternation in membrane potential (62.6 ± 5.0.4% inhibition) and integrity (> 95 ± 2.3%), and the release of UV260 absorbing materials within 60 min (up to 3.2 fold increase, p < 0.01) of exposure. Further, SCL localized to the cytoplasm and hydrolyzed plasmid DNA. While in vitro checkerboard studies revealed that SCL potentiated the antimicrobial activity of topical antimicrobials such as polymixin, neomycin, and bacitracin (Fractional Inhibitory Concentration Index range, 0.26-0.37). Taken together these results suggest that SCL targets the membrane and DNA of MRSA to facilitate its anti-MRSA antimicrobial effect.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.43-48
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• 2017

In a previous study, we analyzed the transcriptome of Scolopendra subspinipes mutilans using next-generation sequencing technology and identified several antimicrobial peptide candidates. One of the peptides, scolopendrasin V, was selected based on the physicochemical properties of antimicrobial peptides using a bioinformatics strategy. In this study, we assessed the antimicrobial activities of scolopendrasin V using the radial diffusion assay and colony count assay. We also investigated the mode of action of scolopendrasin V using flow cytometry. We found that scolopendrasin V's mechanism of action involved binding to the surface of microorganisms via a specific interaction with lipopolysaccharides, lipoteichoic acid, and peptidoglycans, which are components of the bacterial membrane. These results provide a basis for developing peptide antibiotics.

• Food Science and Preservation
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• v.7 no.1
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• pp.1-7
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• 2000

To develop natural antimicrobial agents for keeping qualities of postharvested greenhouse produce the antimiocrobial actions of Polygonum cuspidatum Sieb. et Zucc. extract , which showed remarkable antimicrobial effects against microorganism causing the postharvest decay of greenhouse produce, were investigate. In the inhibitory experiment of enzymes related to energy production metabolism hexokinase activities decreased to 73% and 68% by treating with Polygonum cuspidatum Sieb. et.Zucc. extract and Eugenia caryophyllata Thumnberg extract in comparison with control, respectively. Direct visualization of microbial cells by using both transmission electron microscope and scanning electron microscope showed that microbial cell membrane was destroyed by treating with the dilute extract solution. this change of celluloar membrane permeability could be identified in the experiment that 0-nitrophenyl-${\beta}$-D-galactopyrano-side(ONPG), the artificial substrate of ${\beta}$-galactosidase, was hydrolyzed in the presence of the extract, indicating that the membrane was perturbed. The separation and identification of the most antimicrobialo substances isolated from Polygonum cuspidatum Sieb et. Zucc. extract and Eugenia caryophyllata Thunberg extract were carried out by using gas chromatography and mass spectrometry 9GC/MSD), which were identified as eugenol. As a result, the functionality of Polygonum cuspidatum Sieb. et Zucc. extract and Eugenia caryophyllata Thunberg extract as antimicrobial agents for keeping qualities of postharvested greenhouse produce may be recommended.

• Membrane and Water Treatment
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• v.9 no.6
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• pp.481-487
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• 2018

Membrane clogging or fouling of the membrane caused by organic, inorganic, and biological on the surface is one of the main obstacles to achieve high flux over a long period of the membrane filtration process. So researchers have been many attempts to reduce membrane fouling and found that there is a close relationship between membrane surface hydrophilicity and membrane fouling, such that the same conditions, a greater hydrophilicity were less prone to fouling. Nanotechnology in the past decade is provided numerous opportunities to examine the effects of metal nanoparticles on the both hydrophilic and antibacterial properties of the membrane. In the present study the improvement of hydrophilic and antimicrobial properties of the membrane was evaluated by adding nanoparticles of titanium dioxide and copper oxide. For this purpose, 4% copper oxide and titanium dioxide nanoparticles with a ratio of 0, 30, 50, and 70% of copper oxide added to the polymeric membrane and compare to the pure polymeric membrane. Comparison experiments were performed on E. coli PTCC1998 in two ways disc and tube and also to evaluate membrane hydrophilic by measuring the contact angle and diameter of pores and analysis point SEM has been made. The results show that the membrane-containing nanoparticle has antibacterial properties and its impact by increasing the percentage of copper oxide nanoparticles increases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.3
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• pp.280-286
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• 2021

This study was performed to screen the antimicrobial activities and proteolytic enzyme stability of the acidified extract of the Blue mussel Mytilus edulis. The acidified extract showed potent antimicrobial activities against Gram-positive bacteria, Bacillus subtilis, and Gram-negative bacteria, Escherichia coli D31, but had no activity against Candida albicans. Treatment of extract with trypsin completely abolished all or significant antibacterial activity against the tested bacteria, but slightly decreased antimicrobial activity against B. subtilis, and treatment of extract with chymotrypsin retained almost antibacterial activity against the tested bacteria except for E. coli D31. To confirm the additional enzyme stability of the extract, antimicrobial activity of the extract was tested after treated with several enzymes. Enzymes treated extract showed potent antimicrobial activity against B. subtilis and its activity was also retained for 5 h after trypsin treatments. Non-proteinaceous materials in the acidified extract also showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. All our results indicate that mussel extract might contain the proteinaceous or non-proteinaceous antibacterial materials target not bacterial membrane but intracellular components. These results could be used to develop mussel extract as an additive for the improvement of stability or antimicrobial activity of antibiotics against specific bacteria.