• Journal of the Korean Magnetic Resonance Society
• /
• v.22 no.3
• /
• pp.71-75
• /
• 2018

The CRISPR-Cas system is the adaptive immune system in bacteria and archaea against invading phages or foreign plasmids. In the type II CRISPR-Cas system, an endonuclease Cas9 cleaves DNA targets of phages as directed by guide RNA comprising crRNA and tracrRNA. To avoid targeting and destruction by Cas9, phages employ anti-CRISPR (Acr) proteins that act against host bacterial immunity by inactivating the CRISPR-Cas system. Here we report the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of AcrIIA4 that inhibits endonuclease activity of type II-A Listeria monocytogenes Cas9 and also Streptococcus pyogenesis Cas9 using triple resonance nuclear magnetic resonance spectroscopy. The secondary structures of AcrIIA4 predicted by the backbone chemical shifts show an ${\alpha}{\beta}{\beta}{\beta}{\alpha}{\alpha}$ fold, which is used to determine the solution structure.

• Fisheries and Aquatic Sciences
• /
• v.17 no.2
• /
• pp.215-222
• /
• 2014

To explore marine microorganisms with medical potential, we isolated and identified marine bacteria from floats, marine algae, animals, and sponges collected from Jeju Island, Korea. We isolated and identified 21 different strains from the marine samples by 16S rRNA analysis, cultured them in marine broth, and extracted them with ethyl acetate (EtOAc) to collect secondary metabolite fractions. Next, we evaluated their anti-oxidative and anti-inflammatory effects. Among the 21 strains, the secondary metabolite fraction of Bacillus badius had both strong antioxidant and anti-inflammatory activity, and thus was selected for further experiments. An antioxidant compound detected from the secondary metabolite fraction of B. badius was purified by preparative centrifugal partition chromatography (n-hexane:EtOAc:methanol:water, 4:6:4:6, v/v), and identified as diolmycin A2. Additionally, diolmycin A2 strongly inhibited nitric oxide production. Thus, we successfully identified a significant bioactive compound from B. badius among the bacterial strains collected from Jeju Island.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.22 no.2
• /
• pp.39-49
• /
• 2009

Objective : Phyllostachyos Folium (PF) has been used to treat patients with febrile disease consuming the body fluids marked by fever with restlessness, thirst etc. In the theory of herbology, PF can clear away heat and promote the production of the body fluids, relieve restlessness. Recently PF is known to have anti-bacterial, anti-oxidantic effects. Methods : The present study was carried out to investigate the effects of PF on solid tumor in mice in terms of immune-potentiating and direct cytotoxic action of PF in vitro and vivo study. The present author investigated thymocyte and splenocyte proliferation to confirm immune-potentiating activity of PF and also investigated tumor/body weight ratio and survival rates in tumor bearing mice. Result : In this study, administration of PF decreased tumor/body weight ratio significantly, and prolonged survival duration compared to non-treated control. In addition, treatment with PF suppressed proliferation rate of Sarcoma 180 (S-180) cells significantly, and elevated proliferation rates of thymocytes isolated from normal mice. These results were co-related with in vivo study. Conclusion : In conclusion, these results suggest that PF is useful to treat patient with solid tumor, because PF has direct toxic action for tumor cell and immune -potentiating action for T cells. Further study will need to elucidate exact mechanisms related in anti-cancer action of PF.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.1
• /
• pp.108-114
• /
• 2008

Present study was carried out for development of a new herb product with gastroprotective effect. Natural herbs mentioned that have gastrointenstial protective property in Donggeuibogam and reports were evaluated anti-bacterial activity against Helicobacter pylori, then Ulmus davidiana var. japonica Nakai (YGP) were selected. The material used for the test were water extract of Ulmus davidiana var. japonica Nakai. They were tested individually on HCl ethanol-induced gastric lesion in rats showed the most significant effectiveness, respectively. Then YGP was made, and tested on HCl ethanol, indomethacin-induced gastric lesion, shay ulcer and gastric secretion. Conclusions : YGP showed significant effect on HCl ethanol, indomethacin-induced gastric lesion, and Shay ulcer. It can be regarded that the antigastritic and anti-ulcerative effects of YGP is originated from reduction of total acid output identified by gastric secretion test.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.4
• /
• pp.576-584
• /
• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• Korean Journal of Environmental Biology
• /
• v.35 no.4
• /
• pp.694-705
• /
• 2017

The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

• Journal of dental hygiene science
• /
• v.20 no.4
• /
• pp.213-220
• /
• 2020

Background: The leaves of Perilla frutescens, commonly called perilla and used for food in Korea, contain components with a variety of biological effects and potential therapeutic applications. The purpose of this study was to identify the components of 70% ethanol extracted Perilla frutescens (EEPF) and determine its inhibitory effects on oral microbial activity and production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharides (LPS)-stimulated Raw264.7 macrophages, consequently, to confirm the possibility of using EEPF as a functional component for improving the oral environment and preventing inflammation. Methods: One kg of P. frutescens leaves was extracted with 70% ethanol and dried at -70℃. EEPF was analyzed using high-performance liquid chromatography analysis, and antimicrobial activity against oral microorganisms was revealed using the disk diffusion test. Cell viability was elucidated using a methylthiazolydiphenyl-tetrazolium bromide assay, and the effect of EEPF on LPS-induced morphological variation was confirmed through microscopic observation. The effect of EEPF on LPS-induced production of pro-inflammatory mediators, NO and PGE2 was confirmed by the NO assay and PGE2 enzyme-linked immunosorbent assay. Results: The main component of EEPF was rosemarinic acid, and EEPF showed weak anti-bacterial and anti-fungal effects against microorganisms living in the oral cavity. EEPF did not show toxicity to Raw264.7 macrophages and had inhibitory effects on the morphological variations and production of pro-inflammatory mediators, NO and PGE2 in LPS-stimulated Raw264.7 macrophages. Conclusion: EEPF can be used as a functional material for improving the oral environment through the control of oral microorganisms and for modulating inflammation by inhibiting the production of inflammatory mediators.

• Journal of Plant Biotechnology
• /
• v.34 no.4
• /
• pp.313-322
• /
• 2007

Extract of Japanese red pine needles has been used in Asia pacific regions since long periods believing its valuable properties as tonic and ability of curing diseases of unidentified symptoms. Some selective compounds present in the extract and their effects were analyzed. Carbohydrates and vitamin c were identified using HPLC; terpenoid compounds by GC-MS; anti-bacterial analysis by paper discs, plates count and gastrointestinal motility by whole cell patch clamp. The extract is a mixture of compounds therefore its diverse effect was expected. Self-fermentation in extract proceeds after spontaneous appearance of yeast strains without inoculation. Effects and composition of the extract vary with varying period of self-fermentation. Extract inhibits the growth of bacteria dose dependently exhibiting its antibacterial properties however effectiveness increases with increase in fermentation period. The extract also can modulate gastrointestinal motility in murine small intestine by modulating pace maker currents in ICC mediated through ATP sensitive potassium channel.

• Journal of Life Science
• /
• v.26 no.6
• /
• pp.673-679
• /
• 2016

The development of safe and effective anti-cancer compounds has been seriously required to prevent and treat development of tumor in recent years. Among them, natural compounds derived traditional medicinal stuffs have been paid to attention as an anti-cancer candidate. In this study, aesculetin is a main component of a widely known as a medicinal stuff. It was reported that aesculetin has various biological effects such as anti-inflammatory and anti-bacterial, but its effect related to cell invasion was not discovered. Therefore, in this study, the effect of aesculetin on antioxidant and matrix metalloproteases (MMPs) was investigated in human fibrosarcoma cells, HT1080. First of all, aesculetin showed the scavenging activity of DPPH radical and reducing power in a dose dependent manner. As a result of cytotoxicity, the nontoxic concentration of aesculetin was below 2 μM in HT1080 cells performed by MTT assay. In addition, aesculetin displayed the inhibitory effect on MMP-9 activity related to cell invasion in experiment carried out by gelatin zymography assay. Furthermore, aesculetin increased the expression level of TIMP-1 but decreased the expression level of MMP-9 stimulated with PMA in western blot assay. Furthermore, aesculetin remarkably inhibited cell invasion related to metastasis a dose dependent manner. Above results suggest that aesculetin could exert chemopreventive effect through inhibition of activity and expression of MMP-9 related to cell invasion.

• Microbiology and Biotechnology Letters
• /
• v.45 no.1
• /
• pp.35-42
• /
• 2017

This study aimed to investigate the effects of probiotic fermentation by comparing the skin antioxidant and anti-inflammatory properties of non-fermented (ARE) and fermented (ARE-F) hot water extracts of Agastache rugosa leaves. ARE-F was obtained via ARE fermentation using Lactobacillus rhamnosus HK-9. In vitro, anti-inflammatory properties were evaluated by analyzing the levels of nitric oxide (NO), reactive oxygen species (ROS), and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated HaCaT keratinocytes. In vitro antiradical activity was measured using 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Attenuation of LPS-stimulated NO (p < 0.01), ROS (p < 0.001) and iNOS (p < 0.05) levels by ARE-F was significantly stronger than that by ARE in HaCaT keratinocytes. However, no differences were observed between the DPPH radical scavenging activities of ARE and ARE-F. ARE-F possesses enhanced skin antioxidant and anti-inflammatory properties, suggesting that probiotic bacterial fermentation can be considered an effective tool for augmenting some pharmacological properties of A. rugosa leaves. In brief, the skin antioxidant and anti-inflammatory potentials of A. rugosa leaf extract are augmented by the fermentation with L. rhamnosus HK-9, a probiotic bacterium.