• Title/Summary/Keyword: bacterial secretion system

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The Antimicrobial Peptide CopA3 Inhibits Clostridium difficile Toxin A-Induced Viability Loss and Apoptosis in Neural Cells

  • Yoon, I Na;Hwang, Jae Sam;Lee, Joon Ha;Kim, Ho
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.30-36
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    • 2019
  • Numerous studies have reported that enteric neurons involved in controlling neurotransmitter secretion and motility in the gut critically contribute to the progression of gut inflammation. Clostridium difficile toxins, which cause severe colonic inflammation, are also known to affect enteric neurons. Our previous study showed that C. difficile toxin A directly induces neural cell toxicities, such as viability loss and apoptosis. In the current study, we attempted to identify a potent inhibitor of toxin A-induced neural cell toxicity that may aid in managing toxin A-induced gut inflammation. In our recent study, we found that the Korea dung beetle-derived antimicrobial peptide CopA3 completely blocked neural cell apoptosis caused by okadaic acid or 6-OHDA. Here, we examined whether the antimicrobial peptide CopA3 inhibited toxin A-induced neural cell damage. In neuroblastoma SH-SY5Y cells, CopA3 treatment protected against both apoptosis and viability loss caused by toxin A. CopA3 also completely inhibited activation of the pro-apoptotic factor, caspase-3. Additionally, CopA3 rescued toxin A-induced downregulation of neural cell proliferation. However, CopA3 had no effect on signaling through ROS/p38 $MAPK/p27^{kip1}$, suggesting that CopA3 inhibits toxin A-induced neural cell toxicity independent of this well-characterized toxin A pathway. Our data further suggest that ability of CopA3 to rescue toxin A-induced neural cell damage may also ameliorate the gut inflammation caused by toxin A.

Effects of soybean extracts fermented with Lactic acid bacteria on immune system activity (유산균을 이용한 대두 발효 추출물이 면역계 활성에 미치는 영향)

  • Park, Byung-Doo;Kim, Hye-Ja
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.3
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    • pp.139-153
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    • 2012
  • Objectives : NK cells are spontaneously cytotoxic lymphocytes. These are not only important parts in the first line of defence against bacterial and viral infections of outside, but they may also play a critical role in chronic viral diseases. NK cells kill their targets spontaneously, without the need for prior sensitization and class I MHC restriction by the regulation of cytolytic functions and secretion of a variety of cytokines, such as interleukin-12(IL-12), MCP-1, IL-6, TNF-${\alpha}$, IFN-${\gamma}$. In addition, macrophage and NK cells cooperate through the production of cell mediates. These cooperation and modulation are one of major factors to prevent for evading immune surveillance of cancer. Hence, it could be assumed that if any candidate to enhance activities of macrophage and NK cell, it is considered as a potentially useful agents against cancer. Methods : In our study, to investigate effect of fermented soybean extracts by Lactic acid bacteria (SFE, soybean fermented extracts) work on intestinal immune cell to maintain general immune modulating and anti-cancer activity. We analyzed NK cytotoxicity assay and gene expressions of cytokine related with macrophage and NK cell activity. Results : In vitro experiment, SFE was verified as safety material for cell toxicicty to tumor cell strain without any toxicity of tumor growth inhibition and various cell strain. Effects of macrophage activity stimulating directly by SFE measured induced cytokine. The studies showed that IL-12 production by stimulation of SFE depended on concentration from 0.16mg/mL to 0.63mg/mL with non toxicity to cell, and it was the best activity at 0.63mg/mL. Besides, the effective concentration of SFE producing TNF-${\alpha}$ is similar to IL-12, but it was the best activity at 1.25mg/mL. The level of MCP-1, IL-6 and IFN-${\gamma}$ depended on concentration from 0.16mg/mL to 10mg/mL, IFN-${\gamma}$ showed the best activity at the effective concentration of 0.63mg/mL. With the result of NK cell activity measurement, the spleen cell of mouse injected SFE had 1.5 times higher killing effect than non injected cell. Conclusions : The result of this studies is that Soybean fermetated extracts(SFE) has possibility to immune aided material for the function not only inhibition of microbial infection to macrophage but also activity of adaption immune and cellular immune system.

New Approaches to the Control of Pathogenic Oral Bacteria (바이오필름을 생성하는 병원성 구강 세균을 제어하는 새로운 접근법)

  • Cho, Soo Jeong
    • Journal of Life Science
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    • v.31 no.1
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    • pp.100-108
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    • 2021
  • In the oral cavity, there are hundreds of microbial species that exist as planktonic cells or are incorporated into biofilms. The accumulation and proliferation of pathogenic bacteria in the oral biofilm can lead to caries and periodontitis, which are typical oral diseases. The oral bacteria in the biofilm not only can resist environmental stress inside the oral cavity, but also have a 1,000 times higher resistance to antibiotics than planktonic cells by genes exchange through the interaction between cells in the oral biofilm. Therefore, if the formation of oral biofilm is suppressed or removed, oral diseases caused by bacterial infection can be more effectively prevented or treated. In particular, since oral biofilms have the characteristic of forming a biofilm by gathering several bacteria, quorum sensing, a signaling system between cells, can be a target for controlling the oral biofilm. In addition, a method of inhibiting biofilm formation by using arginine, an alkali-producing substrate of oral bacteria, is used to convert the distribution of oral microorganisms into an environment similar to that of healthy teeth or inhibit the secretion of glucosyltransferase by S. mutans to inhibit the formation of non-soluble glucans. It can be a target to control oral biofilm. This method of inhibiting or removing the oral biofilm formation rather than inducing the death of pathogenic bacteria in the oral cavity will be a new strategy that can selectively prevent or therapeutic avenues for oral diseases including dental caries.

Biochemical Properties and Application of Bacteriocins Derived from Genus Bacillus (Bacillus속 세균 유래 박테리오신의 특성과 응용)

  • Ji-Young Lee;Dae-Ook Kang
    • Journal of Life Science
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    • v.33 no.1
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    • pp.91-101
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    • 2023
  • Bacteriocins are antimicrobial peptides synthesized on ribosomes, produced by bacteria, that inhibit the growth of similar or closely related bacterial strains. Since the discovery of nisin, many bacteriocins with unique structures and various modes of antibacterial activity have been described, and genes encoding production, secretion, and immunity have been reported. Nisin is one of the bacteriocins applied in cheese, liquid eggs, sauces and canned foods. Many of the bacteriocins of the genus Bacillus belong to lantibiotics, which are modified peptides after translation. Other genus Bacillus also produce many non-lantibiotic bacteriocins. Bacteriocins of the genus Bacillus are sometimes becoming more important because of their broader antibacterial spectrum. Bacteriocins are considered attractive compounds in the food and pharmaceutical industries to prevent food spoilage and growth of pathogenic bacteria. Bacteriocins can be used as biological preservatives in a variety of ways in the food system. Biopreservation refers to extending shelf life and improving safety of foods using microorganisms and/or their metabolites. The demand for new antimicrobial compounds has generated great interest in new technologies that can improve food microbiological safety. Applications of bacteriocins are expanding from food to human health. Today, many researchers are shifting their interest in bacteriocins from food preservation to the treatment of bacteria that cause infections and antibiotic-resistant diseases. This exciting new era in bacteriocin research will undoubtedly lead to new inventions and new applications. In this review, we summarize the various properties and applications of bacteriocins produced by the genus Bacillus.

Immuno-stimulatory Activities of a High Molecular Weight Fraction from Cynanchum wilfordii Radix Obtained by Ultrafiltration (한외거르기(Ultrafiltration)에 의하여 분리된 백수오 고분자 분획물의 면역증진 활성)

  • Jang, Mi;Lim, Tae-Gyu;Hong, Hee-Do;Rhee, Young Kyoung;Kim, Kyung-Tack;Lee, Eunjung;Lee, Jeong Hoon;Lee, Yun Ji;Kim, Yeon Bok;Cho, Chang-Won
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.268-274
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    • 2016
  • The purpose of this study was to investigate the immuno-stimulatory activity of the high-molecular-weight fraction (HMWF) of Cynanchum wilfordii (CW) extracts obtained by ultrafiltration in murine macrophage RAW 264.7 cells and to assess its immuno-stimulatory effect in mice. Ultrafiltration was performed with polyethersulfone membranes (30 kDa cutoff) in a cross-flow filtration system to obtain the HMWF of CW. The results showed that the HMWF increased the production of various cytokines such as tumor necrosis factor-${\alpha}$, interleukin-6, and nitric oxide in dose-ependent manners. In addition, HMWF treatment increased the relative spleen weight as well as splenocyte proliferation induced by concanavalin A or bacterial lipopolysaccharide in mice. Natural killer (NK) cell activity in the HMWF-treated group was significantly increased compared to that in the control group. These results suggest that the HMWF of CW can support the immune system through secretion of macrophage cytokines, thereby enhancing NK cell activity and murine splenocyte proliferation.

Quantitative Analysis of Paeoniflorin and Paeonol in Peony Extracts and Quality Control Standards (모란 추출액에서 paeoniflorin과 paeonol 동시 정량 분석 및 화장품 원료의 품질관리 기준 설정)

  • Yun, Ki-Hun;Chi, Yong-Ha;Lee, Dong-Kyu;Paik, Soo-Heui
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.1
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    • pp.235-246
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    • 2018
  • Paeony has pharmacological activities such as anti-inflammatory, anti-allergic, anti-bacterial, central inhibitory, gastric secretion inhibition, and antispasmodic activities. In addition, its antioxidant activity and whitening effect being reported, thus it is being explored as raw materials for cosmetics. We compared the changes in the contents of paeoniflorin and paeonol in Peony extracts, depending on the changes of extracting solvents, temperature and time. The HPLC method was set up for simultaneous analysis, the system suitabilities were confirmed by using the calibration curves and the QC samples for each assay batch. Paeonol was detected only in roots, and paeoniflorin was higher in leaf and flower than root. Higher concentrations of both ingredients were extracted when the root was used after grinding to a suitable size, and when 30% 1,3-butylene glycol was used as the extraction solvent. Also the concentrations tended to increase at higher temperature and longer time, but the increase was gradual at over $75^{\circ}C$ and 4 hours. The ratio of root, leaf and flower was determined to be 2+2+1g/0.5kg of batch, reaching the contents criteria of paeoniflorin and paeonol. Finally, we selected as the best extraction condition when the raw materials are mixed with 2+2+1g/0.5kg and extracted with 30% 1,3-butylene glycol as an extraction solvent at $75^{\circ}C$ for 4 hours, considering both the concentrations of two components and the cost of raw materials and manufacturing process, The extraction units were scaled up to 10 kg under this condition.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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