• Journal of Life Science
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• v.22 no.6
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• pp.788-793
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• 2012

High concentration of antibiotics has been used to treat the outbreak of edwardsiellosis caused by Edwardsiella tarda in aquaculture. However, not all of the bacteria have been killed with high concentrations of antibiotics treatment by the formation of persister cells with a dormant state. The main objective of this study was to kill persister cell using antibiotics with the addition of natural plant compounds. Antibiotics used in this study consist of 100 mg/ml florfenicol and 100 mg/ml amoxicillin. Ten natural plant compounds with persister cell inhibitor activity to E. coli were obtained from Protein Engineering and Systems Biology Lab. of Sungkyunkwan University. The persister cell inhibition activities of those natural plant compounds were evaluated in test tube. Concentrations of the antibiotics were in the ranges of 25~200 ${\mu}g/ml$. The persister cell formation was observed after 16 hours of culture. Persister cells were killed by antibiotics with natural plant compounds. Among ten natural plant compounds, Gynostemma pentaphyllum, Mallotus japonicus, and Orixa japonica showed persister cell formation inhibition activities. The optimal concentrations of G. pentaphyllum, M. japonicus, and O. japonica for the inhibitor of persister cell formation were 100 ${\mu}g/ml$, 100 ${\mu}g/ml$, and 200 ${\mu}g/ml$, respectively. In vivo study was carried out to evaluate the effect of the antibiotics with natural plant compounds using aquacultural fish, olive flounder, as test animals. G. pentaphyllum, M. japonicus, and O. japonica of 30 ${\mu}g/ml$, 10 ${\mu}g/ml$, and 10 ${\mu}g/ml$ with antibiotics reduced cumulative mortalities, showing the effectiveness of persister cell inhibition.

• Journal of the Korean Wood Science and Technology
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• v.52 no.2
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• pp.145-156
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• 2024

Unlike resistant cells, persister cells resist antibiotics due to a decreased cellular metabolic rate and can transition back to normal susceptible cells when the antibiotic is removed. These persister cells contribute to the chronic symptoms of infectious diseases and promote the emergence of resistant strains with continuous antibiotic exposure. Therefore, eliminating persister cells represents a promising approach to significantly enhance antibiotic efficacy. Here, we found that Coicis Semen extract reduced Staphylococcus aureus persister cells at a concentration of 0.5 g/L. Linoleic acid and oleic acid, the major components of Coicis Semen extract, exhibited a comparable reduction in persister cells when combined with three antibiotics: ciprofloxacin, oxacillin, and tobramycin. Conversely, these effects were nullified in the presence of the surfactant Tween 80 (1%), suggesting that the hydrophobic characteristics of linoleic acid and oleic acids play a pivotal role in reducing the number of S. aureus persister cells. Considering the concentration-dependent effects of linoleic acid and oleic acid, the persister-reducing activity of Coicis Semen extract was primarily attributed to these fatty acids. Moreover, Coicis Semen extract, linoleic acid, and oleic acid increased the cell membrane permeability of S. aureus. Interestingly, this effect was counteracted by 1% Tween 80, indicating a close association between the reduction of persister cells and the increase in cell membrane permeability. The identified compounds could thus be used to eliminate persister cells, thereby enhancing therapeutic efficacy and shortening treatment duration. When used in conjunction with antibiotics, they may also mitigate chronic symptoms and significantly reduce the emergence of antibiotic-resistant bacteria.

• Journal of Microbiology and Biotechnology
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• v.23 no.9
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• pp.1293-1303
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• 2013

Antibiotic resistance of soilborne Acinetobacter species has been poorly explored. In this study, norfloxacin resistance of a soil bacterium, Acinetobacter oleivorans DR1, was investigated. The frequencies of mutant appearance of all tested non-pathogenic Acinetobacter strains were lower than those of pathogenic strains under minimum inhibitory concentration (MIC). When the quinolone-resistance-determining region of the gyrA gene was examined, only one mutant (His78Asn) out of 10 resistant variants had a mutation. Whole transcriptome analysis using a RNA-Seq demonstrated that genes involved in SOS response and DNA repair were significantly up-regulated by norfloxacin. Determining the MICs of survival cells after norfloxacin treatment confirmed some of those cells were indeed persister cells. Ten colonies, randomly selected from among those that survived in the presence of norfloxacin, did not exhibit increased MIC. Thus, both the low mutation frequency of the target gene and SOS response under norfloxacin suggested that persister formation might contribute to the resistance of DR1 against norfloxacin. The persister frequency increased without a change in MIC when stationary phase cells, low growth rates conditions, and growth-deficient dnaJ mutant were used. Taken together, our comprehensive approach, which included mutational analysis of the target gene, persister formation assays, and RNA sequencing, indicated that DR1 survival when exposed to norfloxacin is related not only to target gene mutation but also to persister formation, possibly through up-regulation of the SOS response and DNA repair genes.

• Journal of Dairy Science and Biotechnology
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• v.39 no.4
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• pp.145-156
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• 2021

Antibiotics are used in many sectors, including the dairy industry, to prevent bacterial infections in humans, animals, and plants. When bacterial cells are exposed to stressors, such as antibiotic exposure, a subpopulation of the cells becomes dormant. This helps the pathogen to revive and reconstitute its pathogenicity. Thus, eradicating the dormant cells may be an effective strategy to reduce the development of antibiotic resistance in bacteria caused by the abuse of antibiotics. In recent years, a large number of indole-related compounds have been reported to eradicate persister cells. In this review, we provide a summary of the mechanisms of persister cell formation and resuscitation, and the ability of indole and substituted indoles to eradicate persister cells.

• Journal of Dairy Science and Biotechnology
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• v.41 no.4
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• pp.157-162
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• 2023

In the field of microbiology, numerous types of bacteria live dormant to survive stresses such as pasteurization and antibiotics. Some bacteria become 'persisters' by inactivating their ribosomes, allowing them to 'sleep' through stress and revive when the stress has been removed. Under stress, some cells morph into hollow, lifeless structures known as 'cell shells.' In microbiology, these cells have been confused with viable cells in the 'viable but non-culturable cells' phenomenon. Therefore, this review addressed the concept that when revival occurs, the always-viable persister cells revive, instead of the dead cell husks.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.130-136
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• 2021

Persister cell formation and biofilms of pathogens are extensively involved in the development of chronic infectious diseases. Eradicating persister cells is challenging, owing to their tolerance to conventional antibiotics, which cannot kill cells in a metabolically dormant state. A high frequency of persisters in biofilms makes inactivating biofilm cells more difficult, because the biofilm matrix inhibits antibiotic penetration. Fatty acids may be promising candidates as antipersister or antibiofilm agents, because some fatty acids exhibit antimicrobial effects. We previously reported that fatty acid ethyl esters effectively inhibit Escherichia coli persister formation by regulating an antitoxin. In this study, we screened a fatty acid library consisting of 65 different fatty acid molecules for altered persister formation. We found that undecanoic acid, lauric acid, and N-tridecanoic acid inhibited E. coli BW25113 persister cell formation by 25-, 58-, and 44-fold, respectively. Similarly, these fatty acids repressed persisters of enterohemorrhagic E. coli EDL933. These fatty acids were all medium-chain saturated forms. Furthermore, the fatty acids repressed Enterohemorrhagic E. coli (EHEC) biofilm formation (for example, by 8-fold for lauric acid) without having antimicrobial activity. This study demonstrates that medium-chain saturated fatty acids can serve as antipersister and antibiofilm agents that may be applied to treat bacterial infections.

• BMB Reports
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• v.53 no.12
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• pp.611-621
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• 2020

Bacterial endoribonuclease toxins belong to a protein family that inhibits bacterial growth by degrading mRNA or rRNA sequences. The toxin genes are organized in pairs with its cognate antitoxins in the chromosome and thus the activities of the toxins are antagonized by antitoxin proteins or RNAs during active translation. In response to a variety of cellular stresses, the endoribonuclease toxins appear to be released from antitoxin molecules via proteolytic cleavage of antitoxin proteins or preferential degradation of antitoxin RNAs and cleave a diverse range of mRNA or rRNA sequences in a sequence-specific or codon-specific manner, resulting in various biological phenomena such as antibiotic tolerance and persister cell formation. Given that substrate specificity of each endoribonuclease toxin is determined by its structure and the composition of active site residues, we summarize the biology, structure, and substrate specificity of the updated bacterial endoribonuclease toxins.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.115-122
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• 2021

Phenol-soluble modulins (PSMs) are responsible for regulating biofilm formation, persister cell formation, pmtR expression, host cell lysis, and anti-bacterial effects. To determine the effect of psm deletion on methicillin-resistant Staphylococcus aureus, we investigated psm deletion mutants including Δpsmα, Δpsmβ, and Δpsmαβ. These mutants exhibited increased β-lactam antibiotic resistance to ampicillin and oxacillin that was shown to be caused by increased N-acetylmannosamine kinase (nanK) mRNA expression, which regulates persister cell formation, leading to changes in the pattern of phospholipid fatty acids resulting in increased anteiso-C15:0, and increased membrane hydrophobicity with the deletion of PSMs. When synthetic PSMs were applied to Δpsmα and Δpsmβ mutants, treatment of Δpsmα with PSMα1-4 and Δpsmβ with PSMβ1-2 restored the sensitivity to oxacillin and slightly reduced the biofilm formation. Addition of a single fragment showed that α1, α2, α3, and β2 had an inhibiting effect on biofilms in Δpsmα; however, β1 showed an enhancing effect on biofilms in Δpsmβ. This study demonstrates a possible reason for the increased antibiotic resistance in psm mutants and the effect of PSMs on biofilm formation.

• Journal of Microbiology and Biotechnology
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• v.33 no.6
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• pp.715-723
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• 2023

Microbial biofilms are resilient, immune-evasive, often antibiotic-resistant health challenges, and increasingly the target for research into novel therapeutic strategies. We evaluated the effects of a nutraceutical enzyme and botanical blend (NEBB) on established biofilm. Five microbial strains with known implications in chronic human illnesses were tested: Candida albicans, Staphylococcus aureus, Staphylococcus simulans (coagulase-negative, penicillin-resistant), Borrelia burgdorferi, and Pseudomonas aeruginosa. The strains were allowed to form biofilm in vitro. Biofilm cultures were treated with NEBB containing enzymes targeted at lipids, proteins, and sugars, also containing the mucolytic compound N-acetyl cysteine, along with antimicrobial extracts from cranberry, berberine, rosemary, and peppermint. The post-treatment biofilm mass was evaluated by crystal-violet staining, and metabolic activity was measured using the MTT assay. Average biofilm mass and metabolic activity for NEBB-treated biofilms were compared to the average of untreated control cultures. Treatment of established biofilm with NEBB resulted in biofilm-disruption, involving significant reductions in biofilm mass and metabolic activity for Candida and both Staphylococcus species. For B. burgdorferi, we observed reduced biofilm mass, but the remaining residual biofilm showed a mild increase in metabolic activity, suggesting a shift from metabolically quiescent, treatment-resistant persister forms of B. burgdorferi to a more active form, potentially more recognizable by the host immune system. For P. aeruginosa, low doses of NEBB significantly reduced biofilm mass and metabolic activity while higher doses of NEBB increased biofilm mass and metabolic activity. The results suggest that targeted nutraceutical support may help disrupt biofilm communities, offering new facets for integrative combinational treatment strategies.

• Journal of Life Science
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• v.26 no.2
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• pp.265-274
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• 2016

Toxin-antitoxin (TA) systems are ubiquitous genetic modules that are evolutionally conserved in bacteria and archaea. TA systems composed of an intracellular toxin and its antidote (antitoxin) are currently classified into five types. Commonly, activation of toxins under stress conditions inhibits diverse cellular processes and consequently induces cell death or reversible growth inhibition. These effects of toxins play various physiological roles in such as regulation of gene expression, growth control (stress response), programmed cell arrest, persister cells, programmed cell death, phage protection, stabilization of mobile genetic elements or postsegregational killing of plasmid-free cells. Accordingly, bacterial TA systems are commonly considered as stress-responsive genetic modules. However, molecule screening for activation of toxin in TA system is available as development of antimicrobial agents. In addition, cytotoxic effect induced by toxin is used as effective cloning method with antitoxic effect of antitoxin; consequently cells containing cloning vector inserted a target gene can survive and false-positive transformants are removed. Also, TA system is applicable to efficient single protein production in biotechnology industry because toxins that are site-specific ribonuclease inhibit protein synthesis except for target protein. Furthermore, some TA systems that induce apoptosis in eukaryotic cells such as cancer cells or virus-infected cells would have a wide range of applications in eukaryotes, and it will lead to new ways of treating human disease. In this review, we summarize the current knowledge on bacterial TA systems and their applications.