• Title/Summary/Keyword: Bacterial Activity

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Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtilis ZA400 in Kimchi Reveals Its Pertinence to Thrombosis Treatment

  • Ahn, Min-Ju;Ku, Hye-Jin;Lee, Se-Hui;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2090-2099
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    • 2015
  • Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn2+, Ca2+, K2+, and Mg2+), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.

Effects of C-Terminal Residues of 12-Mer Peptides on Antibacterial Efficacy and Mechanism

  • Son, Kkabi;Kim, Jieun;Jang, Mihee;Chauhan, Anil Kumar;Kim, Yangmee
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1707-1716
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    • 2019
  • The development of new antimicrobial agents is essential for the effective treatment of diseases such as sepsis. We previously developed a new short peptide, Pap12-6, using the 12 N-terminal residues of papiliocin, which showed potent and effective antimicrobial activity against multidrug-resistant Gram-negative bacteria. Here, we investigated the antimicrobial mechanism of Pap12-6 and a newly designed peptide, Pap12-7, in which the 12th Trp residue of Pap12-6 was replaced with Val to develop a potent peptide with high bacterial selectivity and a different antibacterial mechanism. Both peptides showed high antimicrobial activity against Gram-negative bacteria, including multidrug-resistant Gram-negative bacteria. In addition, the two peptides showed similar anti-inflammatory activity against lipopolysaccharide-stimulated RAW 264.7 cells, but Pap12-7 showed very low toxicities against sheep red blood cells and mammalian cells compared to that showed by Pap12-6. A calcein dye leakage assay, membrane depolarization, and confocal microscopy observations revealed that the two peptides with one single amino acid change have different mechanisms of antibacterial action: Pap12-6 directly targets the bacterial cell membrane, whereas Pap12-7 appears to penetrate the bacterial cell membrane and exert its activities in the cell. The therapeutic efficacy of Pap12-7 was further examined in a mouse model of sepsis, which increased the survival rate of septic mice. For the first time, we showed that both peptides showed anti-septic activity by reducing the infiltration of neutrophils and the production of inflammatory factors. Overall, these results indicate Pap12-7 as a novel non-toxic peptide with potent antibacterial and anti-septic activities via penetrating the cell membrane.

Isolation and Characterization of an Antibacterial Substance from Rheum palmatum for Treatment of Bacterial Vaginosis (대황으로부터 세균성 질염 치료를 위한 항균성 물질의 분리 및 특성)

  • Jang, Jieun;Kang, Dong-Hee;Yoon, Jaewoo;Kim, Hyun-Soo
    • KSBB Journal
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    • v.32 no.2
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    • pp.133-139
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    • 2017
  • Rheum palmatum has traditionally been used as a preventive agent and medication against fever and infection. The aim of this study was to isolate and characterize an antibacterial substance from R. palmatum that is effective against bacterial vaginosis. A methanol extract from R. palmatum showed antibacterial activity against Lactobacillus vaginalis KC TC 3515, Chryseobacterium gleum KCTC 2904, and Sphingomonas paucimobilis KCTC 2834, which cause bacterial vaginosis. After extraction and pH control of the methanol extract from R. palmatum, we found that acidic and alkaline extracts did not show antibacterial activity. A neutral extract (50 mg/mL) displayed an inhibitory zone of 18 mm on a nutrient agar plate with C. gleum KCTC 2904. Fractions No. 11 and 12 among 41 fractions obtained by silica gel column chromatography produced inhibitory zones of 10 mm on nutrient agar plates with C. gleum KCTC 2904. $R_f0.15$ and $R_f0.17$ spots produced by TLC of fraction No. 11 showed antibacterial activity against C. gleum KCTC 2904. Isolation and purification of the peak at a retention time (Rt) of 9.427 min was achieved by HPLC of $R_f0.29spots$. The peak at Rt 9.427 min showed antibacterial activity against C. gleum KCTC 2904.

Isolation, Characterization, and Use for Plant Growth Promotion Under Salt Stress, of ACC Deaminase-Producing Halotolerant Bacteria Derived from Coastal Soil

  • Siddikee, M.A.;Chauhan, P.S.;Anandham, R.;Han, Gwang-Hyun;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1577-1584
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    • 2010
  • In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

Exploring the Potentiality of Novel Rhizospheric Bacterial Strains against the Rice Blast Fungus Magnaporthe oryzae

  • Amruta, Narayanappa;Kumar, M.K. Prasanna;Puneeth, M.E.;Sarika, Gowdiperu;Kandikattu, Hemanth Kumar;Vishwanath, K.;Narayanaswamy, Sonnappa
    • The Plant Pathology Journal
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    • v.34 no.2
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    • pp.126-138
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    • 2018
  • Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

Anti-bacterial effects of lavender and peppermint oils on Streptococcus mutans (Streptococcus mutans 대한 Lavender와 Peppermint Oil의 항균효과)

  • Park, Chung Mu;Yoon, Hyun Seo
    • Journal of Korean Academy of Oral Health
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    • v.42 no.4
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    • pp.210-215
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    • 2018
  • Objectives: The main objectives of this study were to verify the antibacterial activity of two essential oils, lavender and peppermint, against dental caries and to review their synergistic effect when used in combination. Our results provide basic data for the evaluation of the use of these two substances towards the prevention and cure of dental caries. Methods: The sample solutions of lavender and peppermint oils were prepared in three different concentrations (30%, 50%, and 70% (v/v)) by diluting them with third-distilled water and Tween 20. Streptococcus mutans was selected as the bacterial species for testing. The disk diffusion method was used to measure the antibacterial activity of the sample solutions. For generating growth curves and measuring the number of clusters of the bacterial, the liquid medium-dilution method was used; the absorbance of the medium was measured at 600 nm after 3, 6, 12 and 24 hours. Results: When the antibacterial activity of the oils was tested via the disk diffusion method, the activity improved with increasing concentrations of all the sample solutions of peppermint, lavender, and the blend, but there was no significant difference between them with respect to the type of oil. In the growth curves of S. mutans, growth inhibition was observed after 12 hours. The inhibitory effect of 30% lavender oil on growth was 64.9% and 80.1% after 12 and 24 hours of treatment, respectively whereas that of peppermint oil was 71.3% and 80.1% after 12 and 24 hours of treatment, respectively. The inhibitory effect of the blended oil was 71.9% and 81.0% after 12 and 24 hours of treatment, respectively. Conclusions: Further research is still required in order to determine the efficacy of lavender and peppermint oils, as well as other essential oils, for wider use in preventing dental caries.

Immunosuppressive Activity of an Entomopathogenic Bacteria, Xenorhabdus ehlersii KSY, and Its Application to Enhance Insecticidal Activity of Bacillus thuringiensis (곤충병원세균(Xenorhabdus ehlersii KSY)의 곤충면역 억제 능력과 이를 이용한 Bacillus thuringiensis 의 살충력 증가 효과)

  • Kim, Hyoil;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.58 no.2
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    • pp.101-109
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    • 2019
  • An entomopathogenic bacterium, Xenorhabdus ehlersii KSY, is symbiotic to a nematode, Steinernema longicaudum, and exhibits high entomopathogenic virulence against lepidopteran insects. This study showed that the bacterial pathogenicity is induced by its inhibitory activity against eicosanoid biosynthesis of target insects, resulting in immunosuppression. To be applied for insect pest control, the bacteria should be infected to insect hemocoel. To deliver X. ehlersii to inset hemocoel, Bacillus thuringiensis (Bt) was mixed with the bacteria to breakdown the physical barrier (= midgut epithelium) from midgut lumen to hemocoel. The bacterial mixture significantly enhanced insecticidal activity of Bt only against larvae of Plutella xylostella and Maruca vitrata. For formulation, X. ehlersii cells were freeze-dried and mixed with sporulated Bt cells. The formulated bacterial mixture was applied to semi-field cultivating cabbage crop infested by P. xylostella. The bacterial mixture treatment showed over 95% control efficacy, while Bt alone gave 80% control efficacy. These results suggest that X. ehlersii can be applied to develop a novel insect control agent.

Effects of Dietary Fiber on the Bacterial Enzymes and Putrefactive Metabolite in Aged Rats (주요 식이섬유질원이 첨가된 식이가 노화 흰쥐의 장내효소 및 유해산물에 미치는 영향)

  • 강어진;이상선;양차범;신현경
    • The Korean Journal of Food And Nutrition
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    • v.11 no.5
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    • pp.488-492
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    • 1998
  • This study was performed to investigate the influcence of various dietary fiber sources in Korea for activities of bacterial enzymes (${\beta}$-glucosidase, ${\beta}$-glucuronidase) and amounts of putrefactive product (indole) in aged rats. ${\beta}$-Glucosidase activity in the intestinal content was significantly lower in the seamustard 15% group than in other groups whereas the activity of ${\beta}$-glucuronidase was higher in the mugwort 15% group than other experimental groups. The amount of indole and pH in the intestinal content of aged rats were significantly lower in mugwort groups than in other groups.

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Genomic Features and Lytic Activity of the Bacteriophage PPPL-1 Effective against Pseudomonas syringae pv. actinidiae, a Cause of Bacterial Canker in Kiwifruit

  • Park, JungKum;Lim, Jeong-A;Yu, Ji-Gang;Oh, Chang-Sik
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1542-1546
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    • 2018
  • Bacterial canker in kiwifruit is caused by Pseudomonas syringae pv. actinidiae (Psa). In this study, the bacteriophage PPPL-1 effective against Psa was characterized. Belonging to the Podoviridae family, PPPL-1 was effective against most Psa strains as well as most Pseudomonas syringae pathovars. PPPL-1 carries a 41,149-bp genome with 49 protein coding sequences and is homologous to the previously reported phiPSA2 bacteriophage. The lytic activity of PPPL-1 was stable up to $40^{\circ}C$, within a range of pH 3-11 and under 365 nm UV light. These results indicate that the bacteriophage PPPL-1 might be useful to control Psa in the kiwifruit field.

Production of Phagocyte Activating Supernatants by Olive Flounder (Paralichthys olivaceus) Leucocytes Stimulated with Genomic DNA of Escherichia coli

  • Lee Chan Hwei;Kim Dong Soo;Kim Ki Hong
    • Fisheries and Aquatic Sciences
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    • v.5 no.4
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    • pp.258-262
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    • 2002
  • Effects of Escherichia coli genomic DNA on the production of phagocyte activating supernatants by the head kidney leucocytes isolated from olive flounder (Paralichthys olivaceus) were investigated. Phagocyte activating activity of the supernatants was estimated by. measuring reactive oxygen species (ROS) production in target head kidney phagocytes. All supernatants from olive flounder head kidney leucocytes-stimulated with E. coli DNA induced significantly (P<0.01) higher ROS production from target pagocytes than the unstimulated control supernatant. Maximum enhancement of chemiluminescent response was observed $5.0-10.0{\mu}g\;mL^{-1}$ of bacterial DNA while the increment ability was decreased significantly (P<0.01) at the concentration of $20.0{\mu}mL^{-1}$. The results demonstrate that olive flounder head-kidney leucocytes stimulated with bacterial DNA release a soluble phagocyte activating cytokines capable of enhancing the respiratory burst activity from target phagocytes.