• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.18 no.1
• /
• pp.295-301
• /
• 2017

Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

• Microbiology and Biotechnology Letters
• /
• v.35 no.4
• /
• pp.339-346
• /
• 2007

The MJP1 bacterial strain, which possesses antifungal activity, was isolated from meju and identified as Bacillus subtilis based on its morphological and biochemical properties, as well as its 16S rRNA sequence. Antimicrobial activity was found against various species of Gram-positive bacteria, yeasts, and molds, including food-spoilage microorganisms. The antifungal activity was found to be stable after heat and proteolytic enzyme treatment, and in the pH range of $6.0{\sim}10.0$. The antibacterial activity was stable in the pH range of $6.0{\sim}10.0$, but about 50% of the activity was lost after 24 hr at $30^{\circ}C$. The antibacterial compound was also inactivated by proteolytic enzyme treatment, indicating its proteinaceous nature. The apparent molecular masses of the partially purified antifungal and antibacterial compounds, as indicated by using the direct detection method in Tricine-SDS-PAGE, were approximately 2.4 kDa and 4.5 kDa, respectively. These studies suggest that B. subtilis MJP1 produces two bacteriocin-like substances with antifungal and antibacterial activities.

• Microbiology and Biotechnology Letters
• /
• v.39 no.2
• /
• pp.104-110
• /
• 2011

Actinomycetes, Gram positive soil bacteria, are valuable microorganisms which produce useful secondary metabolites including antibiotics, antiparasitic substances, anti-cancer drugs, and immunosuppressants. Although a major family of actinomycetes, known as streptomycetes, has been intensively investigated at the molecular level for several decades, a potentially valuable and only recently isolated non-streptomycetes rare actinomycetes (NSRA) family has been poorly characterized due to lack of proper genetic manipulation systems. Here we report that a PCR-based genome screening strategy was performed with approximately 180 independently isolated actinomycetes strains to isolate potentially valuable NSRA strains. Thanks to this simple PCR-based genome screening strategy we were able to identify only seven NSRA strains, followed by 16S rRNA sequencing for confirmation. Through further bioassays, one potentially valuable NSRA strain (tentatively named Nocardiopsis species MMBL010) was identified which possessed both antifungal and antibacterial activities, along with the presence of polyketide synthase and non-ribosomal peptide synthase genes. Moreover, Nocardiopsis species MMBL010, which was intrinsically recalcitrant to genetic manipulation, was successfully transformed via E. coli-driven conjugation. These results suggest that PCR-based genome screening, followed by the establishment of an E. coli-driven conjugation system, is an efficient strategy to maximize potentially valuable compounds and their biosynthetic genes from NSRA strains isolated from various environments.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.1
• /
• pp.14-18
• /
• 2009

This study was to determine the inhibitory effect against food borne pathogens of ethanol and water extracts from leaf, stem and root of Orostachys japonicus. On the paper disc assay, no detectable bactericidal activity in the water extracts from leaf, stem and root of Orostachys japonicus and ethanol extracts form stem and root of Orostachys japonicus was shown. However, ethanol extract of Orostachys japonicus leaf showed the highest antimicrobial activity. Minimum inhibitory concentration (MIC) of ethanol extracts was determined to range from 0.05 to 0.1% in leaf of Orostachys japonicus against gram positive bacteria and yeast. Antimicrobial activity of ethanol extracts was stable by heating at $121^{\circ}C$ for 15 min, and not affected by pH $2{\sim}10$ except for B. subtilis. These findings suggest ethanol extract from leaf of Orostachys japonicus may be useful as natural preservative.

• Journal of Life Science
• /
• v.22 no.5
• /
• pp.605-612
• /
• 2012

Spore-forming bacteria are being used as probiotic supplements for human and animal use, due to their low pH stability and ability to survive the gastric barrier. In this study, the BCNU 9028 strain was screened from meju, a Korean fermented soybean food starter. Biochemical and physiological characteristics, as well as 16S rDNA sequence analyses, indicate that this strain belongs to the genus $Bacillus$. $Bacillus$ sp. BCNU 9028 showed a 92% survivability at pH 2.5 and could also withstand 0.3% ox bile. Furthermore, it was postulated that $Bacillus$ sp. BCNU 9028 could prevent biofilm formation and adherence of food-borne pathogens such as $Listeria$ $monocytogenes$, $S.$ $aureus$ and $E.$ $coli$ on the basis of its autoaggregation and coaggregation capacity with food-borne pathogens. It was shown that BCNU 9028 has good abilities to adhere to the intestinal tract from its hydrophobic character (63.3%). The $Bacillus$ sp. BCNU 9028 strain especially elicited antibacterial activity against both Gram-positive and -negative pathogens. These findings suggested that the $Bacillus$ sp. BCNU 9028 strain could be used as a potential probiotic.

• Research in Plant Disease
• /
• v.12 no.3
• /
• pp.189-196
• /
• 2006

Crown gall on lower stem of weeping fig(Ficus benjamina Roxb.) was first observed at Daejon in 2003. Tumors were about 15 cm in size and semi-round with rough surface texture of dark brown color. Two virulent isolates among ten bacteria isolated from the tumor tissues were characterized. Their colonies were convex, glistening, circular with an entire edge, and white or tannish cream in color on potato dextrose agar supplemented with 0.5% $CaCO_3$. They were rod shape with peritrichous flagellae, gram-negative, aerobic growth, oxidase-positive, and grew on D1M agar. The isolates were identified as Agrobacterium larrymoorei and A. tumefaciens based on biochemical and physiological characteristics, fatty acid profiles and substrate utilization patterns. Seedlings of some host plants excepting grapevine produced typical galls two to three weeks after inoculation with cell suspensions of the virulent strains. This is the first report on crown gall of weeping fig in Korea.

• Korean journal of food and cookery science
• /
• v.17 no.3
• /
• pp.218-223
• /
• 2001

This study was conducted to evaluate the functional properties of onion, rosemary and thyme extracts which are the ingredients of onion Kimchi. Onion extract showed a significant difference in antioxidative effect based on peroxide value and thiocyanate method. Antioxidative activity of rosemary extract was similar to those of BHA and BHT. but thyme extract did not show any antioxidative effect.. Onion, rosemary, and thyme extracts showed antimicrobial activities against gram positive bacteria but onion and thyme extracts did not against E. coli, Candida, and molds. Rosemary extract demonstrated a strong activity against L. plantarum which is a major lactobacillus in Kimchi fermentation, and Micrococcus luteus. Onion, rosemary, and thyme extracts showed an effect or hangover relief effect by lowering the alcohol concentration of blood in rat. Blood pressure of the male spontaneous hypertensive rat was suppressed by onion extract after 3 days of feeding, but rosemary and thyme extracts were not effective for lowering blood pressure.

• Fisheries and Aquatic Sciences
• /
• v.23 no.8
• /
• pp.20.1-20.18
• /
• 2020

Background: Olive flounder, Paralichthys olivaceus, is an economically important aquaculture species in Korea. Olive flounders have been heavily damaged by streptococcal infections every year and are treated with antibiotics. However, antibiotic abuse is causing the emergence of resistant strains, and to overcome this, research has shown that new antibiotics must be applied. Tylosin is a relatively safe antibiotic and has good activity against Gram-positive bacteria and mycoplasma. We studied the therapeutic effects and side effects of tylosin on Streptococcus parauberis-infected olive flounder. Methods: After artificial infection of olive flounder with S. parauberis SPOF18J3, an appropriate dose of tylosin was confirmed by intramuscular injection (I.M.) at 2.5, 5, 10, and 15 mg/kg, and oral administration at 10 and 20 mg/kg. After I.M. and oral administration dosing of tylosin, side effects were confirmed by serological analysis, histopathological analysis, and median lethal dose (LD₅₀) analysis at both an appropriate concentration and a high concentration. Statistical analysis was performed using one-way analysis of variance (ANOVA) and Tukey's test (p < 0.05). Results: The appropriate I.M. and oral administration concentration of tylosin administered to olive flounder infected with S. parauberis SPOF18J3 was found to be 10 mg/kg. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were showed not significantly different between the control group and the experimental groups. The histopathologic results showed mild inflammatory responses in muscle and tubular vacuolization and tubular atrophy appeared, but there were no significant differences between the groups. The LD₅₀ was confirmed to be 461 mg/kg. Conclusion: In this study, an effective treatment method was provided by verifying the treatment effects and side effects of tylosin in olive flounder infected with S. parauberis, which can be applied directly to aquaculture sites. In addition, these results may be used as a reference for evaluation required upon request to obtain approval for tylosin antibiotics as fishery antibiotics in Korea. After approval, it is possible that a fishery disease manager will be able to prescribe and sell the antibiotic tylosin.

• The Korean Journal of Mycology
• /
• v.40 no.2
• /
• pp.118-123
• /
• 2012

In order to develop environment friendly fungicide for the control of citrus green mold (Penicillium digitatum) using endophytic bacteria, the 21 bacterial isolates were isolated from citrus leaves in seven different orchards in Jeju Province. Among the 21 bacterial isolates, 5 bacterial isolates presented antifungal activity against green mold pathogen P. digitatum. The CB3 isolate, which showed the most strong antagonistic effect, was selected through opposite culture against the pathogen. The rod-shaped, gram-positive bacterium CB3 was identified as Bacillus velezensis based on morphological, physiological characteristics, 16S rDNA, and gyr A gene sequence analysis. The isolate CB3 showed strong antifungal activity against two citrus postharvest pathogen P. digitatum. Citrus fruits were treated by wound inoculation with P. digitatum pathogen, and the control efficacy of CB3 culture broth was 66.7% ($1{\times}10^8$ cfu/ml). In conclusion, The stability of CB3 and its strong antifungal activity also lead us to believe that it has potential for application as an environment friendly biological control agent.

• Korean Journal of Microbiology
• /
• v.50 no.3
• /
• pp.210-215
• /
• 2014

Pseudomonas nitroreducens TX1 was isolated from a rice field drainage in Taiwan. The bacterium is of special interest because of its capability to use a group of nonionic surfactants such as alkylphenol polyethoxylates even at high concentrations as a sole carbon source. In this study, a small cryptic circular plasmid, pTX1, was characterized from P. nitroreducens TX1. It is 2,286 bp in length with a GC content of 63.3% and harbors three open reading frames, $Rep_{pTX1}$ and functionally unidentified ORF1 and ORF2. The predicted $rep_{pTX1}$ gene product is homologous to Rep proteins of plasmids belonging to the pC194/pUB110 family, which is predominantly found in Gram-positive bacteria and is known to replicate by the rolling-circle mechanism. The copy number of pTX1 was estimated to be about 150 in each cell. Based on the genetic fingerprints and comparison with other plasmids, it is concluded that pTX1 replicates by a rolling circle mechanism which is rarely found for Pseudomonas plasmids.