• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1007-1014
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• 2015

Plectasin, the first defensin extracted from a fungus (the saprophytic ascomycete Pseudoplectania nigrella), is attractive as a prospective antimicrobial agent. The purpose of this study was to establish a bacterium-based production system and evaluate the antimicrobial activity of the resulting plectasin. A gene encoding plectasin, with the codon preference of Escherichia coli, was optimized based on its amino acid sequence, synthesized using genesplicing with overlap extension PCR, and inserted into the expression vector pGEX-4T-1. The fusion protein was expressed in the soluble fraction of E. coli and purified using glutathione Stransferase affinity chromatography. Plectasin was cleaved from the fusion protein with thrombin and purified by ultrafiltration. The purified plectasin showed strong, concentrationdependent antimicrobial activity against gram-positive bacteria, including antibiotic-resistant bacteria, especially penicillin-resistant Enterococcus faecium. This antimicrobial activity was equal to chemically synthesized plectasin and was maintained over a wide range of pH and temperatures. This soluble recombinant expression system in E. coli is effective for producing plectasin at a relatively lower cost, and higher purity and efficiency than prior systems, and might provide a foundation for developing a large-scale production system. Overall, plectasin shows potential as a novel, high-performance, and safe antibiotic for the treatment of refractory diseases caused by drug-resistant bacterial strains.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.596-602
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• 2016

Staphylococcus aureus, like other gram-positive pathogens, has evolved a large repertoire of virulence factors as a powerful weapon to subvert the host immune system, among which alpha-hemolysin (Hla), a secreted pore-forming cytotoxin, plays a preeminent role. We observed a concentration-dependent reduction in Hla production by S. aureus in the presence of sub-inhibitory concentrations of isorhamnetin, a flavonoid from the fruits of Hippophae rhamnoides L., which has little antibacterial activity. We further evaluate the effect of isorhamnetin on the transcription of the Hla-encoding gene hla and RNAIII, an effector molecule in the agr system. Isorhamnetin significantly down-regulated RNAIII expression and subsequently inhibited hla transcription. In a co-culture of S. aureus and lung cells, topical isorhamnetin treatment protected against S. aureus-induced cell injury. Isorhamnetin may represent a leading compound for the development of anti-virulence drugs against S. aureus infections.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.189-193
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• 2008

A Gram-positive and endospore-forming strain, $JH8^T$, was isolated from deep-sea sediment and identified as a member of the genus Paenibacillus on the basis of 16S rRNA gene sequence and phenotypic analyses. According to a phylogenetic analysis, the most closely related species was Paenibacillus wynnii LMG $22176^T$ (96.9%). Strain $JH8^T$ was also facultatively anaerobic and grew optimally at $20-25^{\circ}C$. The major cellular fatty acid was anteiso-$C_{15:0}$, and the DNA G+C content was 53.1mol%. The DNA-DNA relatedness between the isolate and Paenibacillus wynnii LMG $22176^T$ was 7.6%, indicating that strain $JH8^T$ and P. wynnii belong to different species. Based on the phylogenetic, phenotypic, and chemotaxonomic characteristics, strain $JH8^T$ would appear to belong to a novel species, for which the name Paenibacillus donghaensis sp. novo is proposed (type strain=KCTC $13049^T=LMG\;237S0^T$).

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.932-945
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• 2009

Several species of Gram-positive bacteria have cell wall peptidoglycan (syn. murein) in which not all of the sugar moieties are N-acetylated. This has recently been shown to be a secondary effect, caused by the action of a peptidoglycan N-acetylglucosamine deacetylase. We have found that the opportunistic pathogen Listeria monocytogenes is unusual in having three enzymes with such activity, two of which remain in the cytoplasm. Here, we examine the enzyme (PgdA) that crosses the cytoplasmic membrane and is localized in the cell wall. We purified a hexa-His-tagged form of PgdA to study its activity and constructed a mutant devoid of functional Lmo0415 (PgdA) protein. L. monocytogenes PgdA protein exhibited peptidoglycan N-acetylglucosamine deacetylase activity with natural substrates (peptidoglycan) from both L. monocytogenes and Escherichia coli as well as the peptidoglycan sugar chain component N-acetylglucosamine, but not with N-acetylmuramic acid. As was reported recently [6], inactivation of the structural gene was not lethal for L. monocytogenes nor did it affect growth rate or morphology of the cells. However, the pgdA mutant was more prone to autolysis induced by such agents as Triton X-100 and EDTA, and is more susceptible to the cationic antimicrobial peptides (CAMP) lysozyme and mutanolysin, using either peptidoglycan muramidases or autolysis-inducing agents. The pgdA mutant was also slightly more susceptible than the wild-type strain to the action of certain beta-lactam antibiotics. Our results indicate that protein PgdA plays a protective physiological role for listerial cells.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1841-1847
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• 2007

We previously identified the origin of replication of p703/5, a small cryptic plasmid from the KBL703 strain of Enterococcus faecalis. The origin of replication contains putative regulatory cis-elements required for replication and a replication initiator (RepA) gene. The replicon of p703/5 is similar in its structural organization to theta-type plasmids, and RepA is homologous to a family of Rep proteins identified in several plasmids from Gram-positive bacteria. Here, we report molecular interactions between RepA and the replication origin of p703/5. DNase I footprinting using recombinant RepA together with electrophoretic mobility shift assays confirmed the binding of RepA to the replication origin of p703/5 via iterons and an inverted repeat. We also demonstrated the formation of RepA dimers and the different binding of RepA to the iteron and the inverted repeat using gel filtration chromatographic analysis, a chemical crosslinking assay, and electrophoretic mobility shift assays in the presence of guanidine hydrochloride. Our results suggest that RepA plays a regulatory role in the replication of the enterococcal plasmid p703/5 via mechanisms similar to those of typical iteroncarrying theta-type plasmids.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1198-1205
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• 2016

Lactic acid bacteria (LAB) have beneficial effects on intestinal health and skin diseases. Lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria, is known to induce the production of several cytokines such as TNF-α, IL-1β, and IL-8 and affect the intestinal microflora, anti-aging, sepsis, and cholesterol level. In this study, Weissella cibaria was isolated from Indian dairy products, and we examined its immune-enhancing effects. Live and heat-killed W. cibaria did not induce the secretion of immune-related cytokines, whereas LTA isolated from W. cibaria (cLTA) significantly increased the secretion of TNF-α, IL-1β, and IL-6 in a dose-dependent manner. cLTA increased the phosphorylation of nuclear factor kappa-light-chain-enhancer of activated B cells, p38 mitogen-activated protein kinases, and c-Jun N-terminal kinases in THP-1 cells. The secretion of TNF-α and IL-6 was also increased in the cLTA-treated mouse splenocytes. These results suggest that cLTA, but not W. cibaria whole cells, has immune-boosting potential and can be used to treat immunosuppression diseases.

• Journal of Microbiology and Biotechnology
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• v.23 no.10
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• pp.1357-1364
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• 2013

Human skin is continuously exposed to ultraviolet (UV)-induced photoaging. UVA increases the activity of MMP-1 in dermal fibroblasts through mitogen-activated protein kinase (MAPK), p38, signaling. The irradiation of keratinocytes by UVA results in the secretion of the inflammatory cytokine, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and the stimulation of MMP-1 in normal human dermal fibroblasts (NHDFs). Lipoteichoic acid (LTA) is a component of the cell wall of gram-positive Lactobacillus spp. of bacteria. LTA is well known as an anti-inflammation molecule. LTA of the bacterium Lactobacillus plantarum has an anti-photoaging effect, but the potential anti-photoaging effect of the other bacteria has not been examined to date. The current study showed that L. sakei LTA (sLTA) has an immune modulating effect in human monocyte cells. Our object was whether inhibitory effects of sLTA on MMP-1 are caused from reducing the MAPK signal in NHDFs. It inhibits MMP-1 and MAPK signaling induced by UVA in NHDFs. We also confirmed effects of sLTA suppressing TNF-${\alpha}$ inducing MMP-1 in NHDFs.

• Journal of Fisheries and Marine Sciences Education
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• v.27 no.1
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• pp.13-17
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• 2015

Oxytetracycline (OTC) has been widely used in aquaculture field as a therapeutic and prophylactic agent because of its broad-spectrum activity against gram-positive and negative bacteria. Residual oxytetracycline (OTC) was studied after spray treatment of cultured abalone, Haliotis discus hannai. Muscle concentration of OTC was determined after spray treatment ( 4,000, 7,000, 10,000 ppm) in the abalone. Muscle samples were taken at 0, 1, 3, 7, 14, 21 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330 and 360 day post-dose. OTC analyses were carried out by high performance liquid chromatography (HPLC). In 4000, 7000 and 1000 ppm treatment for small size abalones, OTC levels at 1 day post-dose, have been dramatically decreased by 8.34, 3.35 and 4.47 ug/g, respectively. For medium size abalones, concentration were measured as 7.58, 15.62 and 7.8 ug/g, respectively. Those of large size abalones also were observed as 11.31, 12.38 and 15.28 ug/g, respectively, at 1 day post-dose. No significant differences in residual OTC depletion in muscles were observed between the size of abalones. More than 0.2 mg/kg of OTC was detected in muscle tissues and the residues were found over 60 days after treatment. It is expected that these results would contribute to improve recommended withdrawl periods of OTC for a safer seafood supply.

• The Korean Journal of Mycology
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• v.12 no.3
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• pp.85-92
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• 1984

To isolate antibiotic-producing microorganisms from Korean soils, microbes were isolated from the soil samples and screened for antibacterial activity. A strain which was isolated from the soil sample collected in Choong Chung Book Do had a high antibacterial activity against gram-positive bacteria. The examination of morphological and physiological characteristics of that strain according to the International Streptomyces Project methods showed that it was one of Streptomyces species. After the antibacterial constituent of the strain was produced in submerged culture method, it was isolated and purified by XAD-2 and CM-Sephadex column chromatography. And it was found to be one of quinone type antibiotics.

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.265-273
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• 2009

A cross-sectional survey was undertaken to identify risk factors and clinical signs associated with parasitic helminth infections of cattle in Mymensignh district of Bangladesh. A nonrandom convenience sampling method was used to select 138 animals from 40 farmers/herds. The eggs per gram of faeces (epg) for nematodes and trematodes were determined by McMaster and Stoll's methods respectively. Animal-level and herd-level data were recorded by means of a questionnaire. Multi-collinearity amongst explanatory variables were assessed using $2{\times}2{\times}\;X^2$ test and one variable in a pair was dropped if $P{\leq}0.05$ formultiple logistic regression models. Association study between outcome and explanatory variables was conducted using classification tree, random forests and multiple logistic regression. A positive epg was considered as infected. Analyses were performed using $STATA^{(R)}$, version 8.0/Intercooled and $R^{(R)}$, Version 2.3.0. Seventy eight percent of the cattle were found to be infected with at least one type of helminth. Twenty four pairs of combinations of explanatory variables showed significant associations. Male animals (OR=3.3, P=.006, 95% CI=1.4, 7.7) were associated with significantly increased prevalence of nematode infection. Female cattle of the study area are mostly cross-breed, kept indoor, fed relatively good diet and not used for draught purpose. Males are used for draught purpose thereby more exposed to nematode infective stage and provided with relatively poor diet. So stressed male cattle may become more susceptible to nematode infection. All of the three statistical techniques selected gender and lumen motility as most important variables in association with nematode infection in cattle. The result of this survey can only be extrapolated to the periurban cattle population of traditional management system.