• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.2
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• pp.137-147
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• 2017

The human skin is an ecosystem that provides habitat to various microorganisms. These comprise the skin microbiome and provide numerous benefits in addition to maintaining a symbiotic relation with the host. Various metabolites generated by the skin microbiome exert beneficial effects such as strengthening the skin barrier, and anti-aging and anti-inflammatory functions. In this study, we isolated a novel bacterium, designated Sporichthyacae strain K-07, from the human skin. Analysis of 16S rRNA gene sequences showed that the newly found bacterium shares 93.4% homology with the genus Sporichthya, thus corroborating the discovery of a novel genus. We further analyzed the effect of the novel strain in vitro, by treating HaCaT cells with bacterial metabolite products. Treatment resulted in changes in the mRNA expression levels of filaggrin, claudin1, claudin4, SMase, CERS3, HAS3, aquaporin3, IL-6, TNF-${\alpha}$, TSLP, and TARC. Specifically, the levels of filaggrin, claudin1, claudin4, SMase, CERS3, HAS3, and aquaporin3 were higher in strain K-07 metabolite product-treated cells than in control cells. These results showed that metabolite products of the novel strain K-07 enhanced the skin barrier and exert anti-inflammatory effects. Therefore, these metabolite products could be potentially used for treatment of skin conditions.

• Journal of Life Science
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• v.25 no.7
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• pp.833-838
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• 2015

Multidrug-resistant super bacterial, fungal, viral, and parasitic infections are major health threaten pathogens. However, to overcome the present healthcare situation, among the leading alternatives to current drugs are antimicrobial peptides (AMPs), which are abundantly produced via various species in nature. AMPs, small host defense proteins, are in charge of the innate immunity for the protection of multicellular organisms such as fish, amphibian, reptile, plants and animals from infection. The number of AMPs identified per year has increased steadily since the 1980s. Over 2,000 natural AMPs from bacteria, protozoa, fungi, plants, and animals have been listed into the antimicrobial peptide database (APD). The majority of these AMPs (>86%) possess 11–50 amino acids with a net charge from 0 to +7 and hydrophobic percentages between 31–70%. This report classified AMP into several categories including biological source, biological functions, peptide properties, covalent bonding pattern, and 3D structure. AMP functions not only antimicrobial activity but facilitates cell biological activity such as chemotatic activity. In addition, fibroblastic reticular cell (FRC) originated from mouse lymph node stroma induced the expression of AMP in inflammatory condition. AMP induced from FRC contained whey acidic protein (WAP) domain. It suggests that the classification of AMP will be done by protein domain.

• Journal of Life Science
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• v.24 no.3
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• pp.219-226
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• 2014

The immune system protects the body from bacterial infection and disease, as well as cancer that develops following the mutation of cells. Aging exerts adverse effects on the immune system, such as chronic inflammation, resulting in rheumatoid arthritis. The purpose of this study was to investigate the anti-inflammatory effectiveness of Scutellaria baicalensis, which contains baicalin. HPLC analysis showed that S. baicalensis hot water extracts (SBWE) contained 42.2 mg/g of baicalin. To evaluate the cytotoxicity of SBWE, an MTT assay was carried out in Raw264.7 cells. No cytotoxicity was observed below 160 ${\mu}g/ml$ of SBWE. SBWE at 40 ${\mu}g/ml$ reduced the amount of nitric oxide produced by macrophages stimulated with lipopolysaccharide by 40%. In addition, SBWE inhibited phagocytosis stimulated with zymosan. Furthermore, the content of tumor necrosis factor-alpha ($TNF-{\alpha}$) produced by the macrophages was decreased in the presence of SBWE in a dose-dependent manner. SBWE also inhibited the production of interleukin-1 beta ($IL-1{\beta}$) in a time course experiment. Moreover, treatment with 20 ${\mu}g/ml$ of SBWE remarkably decreased the expression level of cyclooxynase-2. The results provide evidence that SBWE may exert an anti-inflammatory effect through modulation of the immune system.

• The Korean Journal of Pesticide Science
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• v.15 no.4
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• pp.485-489
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• 2011

In previous reports, the treatment of Bacillus amyloliquefaciens strain EXTN-1 showed a broad diseasecontrolling spectrum to the plant diseases caused by viral, bacterial, and fungal pathogens as well as the promotion of plant growth. In mechanisms of EXTN-1, treatment of EXTN-1 increased oxidative burst in early stage and induced the expression of resistance genes, PR-1a, PDF1.2. Mechanism involved in induced systemic resistance by EXTN-1 was revealed as simultaneous activation of SA and JA or ethylene metabolic pathways. The purpose of this study was to determine whether B. amyloliquefaciens EXTN-1 has a similar effect on rice plant against Rice stripe tenuivirus (RSV) under greenhouse conditions. When rice seeds were soaked in B. amyloliquefaciens strain EXTN-1, rice plants showed significant systemic resistance against RSV as well as promoted growth. In the case of plant growth, in 30-day old plants treated with B. amyloliquefaciens EXTN-1, the heights, weights, and lengths of roots increased by 12.6%, 9.8%, and 16.0%, respectively confirming the effects of PGPR. When the induced systemic resistance to RSV was examined, in 20-day old plants were treated with B. amyloliquefaciens EXTN-1, the heights, weights, and lengths of roots increased by 8.4%, 10.9%, and 4.8%, respectively compared to the control. Induced systemic resistance was more prominent in susceptible cultivars - Chucheong and Ilpum compared to the resistant cultivar, Nakdong.

• Journal of Korean Society of Forest Science
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• v.79 no.3
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• pp.278-284
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• 1990

We have demonstrated expression of bacterial genes transferred into cells of Populus nigra ${\times}$ P. maximowiczii by A. tumefaciens strain 6044 (pGA 472). We determined the optimum concentration of kanamycin sulfate for effective selection of punctured leaf transformed using Agrobacterium binary vector pGA 472 containing a neomycine phosphotransferase gene (NPT-II) which confers kanamycin resistance. The combination of cefotaxime (200mg/l) and carbenicillin (300mg/l) showed good performance of discarding Agrobacterium from inoculated punctured leaf. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited callus and shoots induction from punctured leaf. Number of shoots regenerated from co-cultured punctured leaf was 3.0 on MS basal medium supplemented with 10 mg/l kanamycin sulfate, while that of not co-cultured punctured leaf was none. The regeneration rate was 10% from the punctured leaf co-cultured on MS medium with 10 mg/l kanamycin. Regenerated shoots are developing from micropropagation for Southern blot analysis and inheritance of the kanamycin resistance trait (NPT-II).

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.291-299
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• 2009

To elucidate HtrA3's functional roles in the HtrA3 mediated cellular processes, it is necessary to investigate its biochemical characteristics. In the present study, we constructed the plasmids encoding putative mature HtrA3 proteins (M1-HtrA3 and M2-HtrA3) based on the putative maturation sites of highly homologous HtrA1 and mouse HtrA3. We used the pGEX bacterial expression system to develop a simple and rapid purification for the recombinant HtrA3 protein. Although yields of the mature HtrA3 proteins were slightly low as 10~50 ${\mu}g$/L, the amounts and purity of M1- and M2-HtrA3 were enough to investigate their proteolytic activities. The putative mature HtrA3 proteins have proteolytic activity which could cleave $\beta$-casein as an exogenous substrate. We compared the proteolytic activity between the HtrA family, HtrA1, HtrA2, and HtrA3. The cleavage activity of HtrA3 and HtrA2 were 2 folds higher than that of HtrA1, respectively. Our study provides a method for generating useful reagents to identify natural substrates of HtrA3 in the further studies.

• Microbiology and Biotechnology Letters
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• v.40 no.3
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• pp.274-277
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• 2012

Cytokinesis is the final stage of cell division, dividing one mother cell into two daughter cells. For the cutting of a plasma membrane during bacterial cytokinesis, a tubulin homolog FtsZ protein is recruited from the cytoplasm to the division site. FtsZ protein polymerizes in a GTP-dependent manner and its N-terminal domain has a GTPase activity. In this study, we have begun to characterize FtsZ from Staphylococcus aureus (SA). Full-length SA FtsZ was cloned into pRSFDuet-1 vector and the clone was transformed into a BL21 (DE3) star cell. The recombinant SA FtsZ protein was purified using Ni-NTA affinity chromatography and dialysis. Using a spectrofluorometer, we showed that SA FtsZ undergoes a GTP-dependant polymerization in vitro. The polymer of the SA FtsZ protein disappeared after a few minutes, suggesting that the polymer is degraded as the GTP is consumed. This assay system may well be applied for inhibitor screening targeting S. aureus FtsZ.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.3
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• pp.323-328
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• 2019

Tuberculosis, a chronic bacterial infection caused by Mycobacterium tuberculosis (MTB), differs in its status latency and activity because of the characteristics of MTB, immune status of the host, and genetic susceptibility. The host defense mechanism against MTB is caused mainly by interactions between macrophages, T cells, and dendritic cells. CD44 is expressed in activated T cells when infected with MTB and regulates lymphocyte migration. In addition, CD44 mediates leukocyte adhesion to the ECM and plays a role in attracting macrophages and $CD4^+$ T cells to the lungs. Therefore, genetic polymorphism of the CD44 gene will inhibit the host cell immune mechanisms against MTB. This study examined whether the genetic polymorphism of the CD44 gene affects the susceptibility of tuberculosis. A total of 237 SNPs corresponding to the CD44 genes were analyzed using the genotype data of 443 tuberculosis cases and 3,228 healthy controls from the Korean Association Resource (KARE). Of these, 17 SNPs showed a significant association with the tuberculosis case. The most significant SNP was rs75137824 (OR=0.231, CI: 1.51~3.56, $P=1.3{\times}10^{-4}$). In addition, rs10488809, one of the 17 significant SNPs, is important for the tuberculosis outbreak can bind to the JUND and FOS transcription factors and can affect CD44 gene expression. This study suggests that polymorphism of the CD44 gene modulates the host susceptibility to tuberculosis in a variety of ways, resulting in differences in the status of tuberculosis.

• Journal of Life Science
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• v.31 no.1
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• pp.83-89
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• 2021

Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.

• Journal of Food Hygiene and Safety
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• v.35 no.6
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• pp.535-543
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• 2020

In addition to the ubiquitous roles of cellular RNA in genetic regulations, gene expression and phenotypic variations in response to environmental cues and chemotactic signals, the regulatory roles of a new type of RNA called extracellular RNAs (exRNAs) are an up-and-coming area of research interest. exRNA is transported outside the cell through membrane blebs known as membrane vesicles or extracellular vesicles (EVs). EV formation is predominant and conserved among all microbial forms, including prokaryotes, eukaryotes, and archaea. This review will focus on the three major topics concerning bacterially derived exRNAs, i.e., 1) the discovery of exRNA and influence of extraneous RNA over bacterial gene regulations, 2) the known secretion mechanism for the release of exRNA, and 3) the possible applications that can be devised with these exRNA secreted by different gram-negative and gram-positive bacteria. Further, this review will also provide an opinion on exRNA- and EV-derived applications such as the species-specific exRNA markers for diagnostics and the possible roles of exRNA in probiotics and the epigenetic regulations of the gut microbiome.