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한국미생물생명공학회(구 한국산업미생물학회)> Journal of Microbiology and Biotechnology> Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells

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Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells

Yong-shi Liu , Qiong Liu , Yan-long Jiang , Wen-tao Yang , Hai-bin Huang , Chun-wei Shi , Gui-lian Yang , Chun-feng Wang
  • : 한국미생물생명공학회(구 한국산업미생물학회)
  • : Journal of Microbiology and Biotechnology 30권4호
  • : 연속간행물
  • : 2020년 04월
  • : 515-525(11pages)

DOI


목차

1. Introduction
Materials and Methods
Results
Discussion
Acknowledgments
Conflict of Interest
References

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초록 보기

Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.

UCI(KEPA)

간행물정보

  • : 자연과학분야  > 생물
  • : KCI등재
  • : SCI,SCOPUS
  • : 월간
  • : 1017-7825
  • : 1738-8872
  • : 학술지
  • : 연속간행물
  • : 1991-2020
  • : 5533


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1Potential Health-Promoting Benefits of Paraprobiotics, Inactivated Probiotic Cells

저자 : Shahina Akter , Jong-hyun Park , Hoo Kil Jung

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 477-481 (5 pages)

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Viability plays an important role in the beneficial microbes (probiotics) to produce health benefits. However, this idea has been changed after the invention of the term "paraprobiotics," indicating that non-viable microbes could produce health benefits similar to those produced by live probiotics. Occasionally, it might be dangerous to administer live probiotics to people with weak immunity. In such cases, ingestion of paraprobiotics could be a potential alternative. The definition of paraprobiotics refers to the use of inactivated (non-viable) microbial cells or cell fractions to provide health benefits to the consumer. Paraprobiotics have attracted much attention because of their long shelf life, safety, and beneficial effects, such as modulation of immunity, modification of biological responses, reduction of cholesterol, anti-inflammatory, and antiproliferative properties. These features indicate that paraprobiotics may play a vital role in improving the health of the consumer by enhancing particular physiological functions, even though the exact underlying mechanisms have not yet been completely elucidated. In this mini-review, we briefly discuss the historical backgrounds of paraprobiotics and evidence of their health-promoting effects, prophylactic, and therapeutic properties.

2β-Sitosterol Contributes in the Resistance to Invasion and Survival of Brucella abortus 544 within RAW264.7 Cells, and Cytokine Production with Reduced Susceptibility to Infection in BALB/c Mice

저자 : Alisha Wehdnesday Bernardo Reyes , Lauren Togonon Arayan , Tran Xuan Ngoc Huy , Son Hai Vu , Wongi Min , Jin Hur , Suk Kim

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 482-489 (8 pages)

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We previously identified β-sitosterol (BS) as one of the most abundant compounds found in Korean red ginseng oil. BS is a widely prevalent vegetable-derived phytosterol with many known health benefits. Here, we investigated the efficacy of BS against Brucella (B.) abortus infection. BS showed no effect on bacterial growth but attenuated internalization, intracellular survival and MAPKs-linked intracellular signaling in RAW264.7 cells. BS treatment in cells is also associated with increased nitrite concentration during infection at 24 h. Slightly enhanced resistance to B. abortus infection was observed in mice orally given BS, which could be mediated by induced production of proinflammatory cytokines. Taken together, our study demonstrates the contribution of BS treatment against B. abortus infection although further investigation is encouraged to maximize its beneficial effects against intracellular infection.

3Croton hirtus L'Hér Extract Prevents Inflammation in RAW264.7 Macrophages Via Inhibition of NF-κB Signaling Pathway

저자 : Min Jeong Kim , Ju Gyeong Kim , Kong Many Sydara , Sang Woo Lee , Sung Keun Jung

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 490-496 (7 pages)

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Consumption of anti-inflammatory nutraceuticals may help treat or prevent inflammation-related illnesses such as diabetes, cardiovascular disease, and cancer. This study evaluated the effect of Croton hirtus L'Hér extract (CHE) on lipopolysaccharide (LPS)-induced nitric oxide (NO) production and nuclear factor kappa-B (NF-κB) signaling cascades. CHE significantly suppressed LPS-induced NO production and inducible nitric oxide synthase (iNOS) expression in RAW264.7 macrophages, although cyclooxygenase (COX)-2 expression was not affected. CHE also suppressed LPS-induced IκB kinase (IKK), IκB, and p65 phosphorylation in RAW264.7 cells. Western blot and immunofluorescence assays of cytosol and nuclear p65 and the catalytic subunit of NF-κB showed that CHE suppressed LPS-induced p65 translocation from the cytosol to the nucleus. CHE also suppressed LPS-induced Interleukin (IL)-6 and tumor necrosis factor (TNF)-α production in RAW264.7 cells. These results suggest that CHE prevents NO-mediated inflammation by suppressing NF-κB and inflammatory cytokines.

4Brucella melitensis omp31 Mutant Is Attenuated and Confers Protection Against Virulent Brucella melitensis Challenge in BALB/c Mice

저자 : L Verdiguel-fernández , R Oropeza-navarro , Adolfo Ortiz , Mg Robles-pesina , J Ramírez-lezama , A Castañeda-ramírez , A Verdugo-rodríguez

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 497-504 (8 pages)

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For control of brucellosis in small ruminants, attenuated B. melitensis Rev1 is used but it can be virulent for animals and human. Based on these aspects, it is essential to identify potential immunogens to avoid these problems in prevention of brucellosis. The majority of OMPs in the Omp25/31 family have been studied because these proteins are relevant in maintaining the integrity of the outer membrane but their implication in the virulence of the different species of this genus is not clearly described. Therefore, in this work we studied the role of Omp31 on virulence by determining the residual virulence and detecting lesions in spleen and testis of mice inoculated with the B. melitensis LVM31 mutant strain. In addition, we evaluated the conferred protection in mice immunized with the mutant strain against the challenge with the B. melitensis Bm133 virulent strain. Our results showed that the mutation of omp31 caused a decrease in splenic colonization without generating apparent lesions or histopathological changes apparent in both organs in comparison with the control strains and that the mutant strain conferred similar protection as the B. melitensis Rev1 vaccine strain against the challenge with B. melitensis Bm133 virulent strain. These results allow us to conclude that Omp31 plays an important role on the virulence of B. melitensis in the murine model, and due to the attenuation shown by the strain, it could be considered a vaccine candidate for the prevention of goat brucellosis.

5Elucidation of the Biosynthetic Pathway of Vitamin B Groups and Potential Secondary Metabolite Gene Clusters Via Genome Analysis of a Marine Bacterium Pseudoruegeria sp. M32A2M

저자 : Sang-hyeok Cho , Eunju Lee , So-ra Ko , Sangrak Jin , Yoseb Song , Chi-yong Ahn , Hee-mock Oh , Byung-kwan Cho , Suhyung Cho

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 505-514 (10 pages)

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The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and nonribosomal proteins.

6Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells

저자 : Yong-shi Liu , Qiong Liu , Yan-long Jiang , Wen-tao Yang , Hai-bin Huang , Chun-wei Shi , Gui-lian Yang , Chun-feng Wang

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 515-525 (11 pages)

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Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.

7Acinetobacter pullorum sp. nov., Isolated from Chicken Meat

저자 : Arxel G. Elnar , Min-gon Kim , Ju-eun Lee , Rae-hee Han , Sung-hee Yoon , Gi-yong Lee , Soo-jin Yang , Geun-bae Kim

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 526-532 (7 pages)

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A bacterial strain, designated B301T and isolated from raw chicken meat obtained from a local market in Korea, was characterized and identified using a polyphasic taxonomic approach. Cells were gram-negative, non-motile, obligate-aerobic coccobacilli that were catalase-positive and oxidase-negative. The optimum growth conditions were 30°C, pH 7.0, and 0% NaCl in tryptic soy broth. Colonies were round, convex, smooth, and cream-colored on tryptic soy agar. Strain B301T has a genome size of 3,102,684 bp, with 2,840 protein-coding genes and 102 RNA genes. The 16S rRNA gene analysis revealed that strain B301T belongs to the genus Acinetobacter and shares highest sequence similarity (97.12%) with A. celticus ANC 4603T and A. sichuanensis WCHAc060041T. The average nucleotide identity and digital DNA-DNA hybridization values for closely related species were below the cutoff values for species delineation (95–96% and 70%, respectively). The DNA G+C content of strain B301T was 37.0%. The major respiratory quinone was Q-9, and the cellular fatty acids were primarily summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:0, and C18:1 ω9c. The major polar lipids were phosphatidylethanolamine, diphosphatidyl-glycerol, phosphatidylglycerol, and phosphatidyl-serine. The antimicrobial resistance profile of strain B301T revealed the absence of antibiotic-resistance genes. Susceptibility to a wide range of antimicrobials, including imipenem, minocycline, ampicillin, and tetracycline, was also observed. The results of the phenotypic, chemotaxonomic, and phylogenetic analyses indicate that strain B301T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter pullorum sp. nov. is proposed. The type strain is B301T (=KACC 21653T = JCM 33942T).

8Biokinetics of Protein Degrading Clostridium cadaveris and Clostridium sporogenes in Batch and Continuous Mode of Operations

저자 : Taewoan Koo , Md Abu Hanifa Jannat , Seokhwan Hwang

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 533-539 (7 pages)

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A quantitative real-time polymerase chain reaction (QPCR) was applied to estimate biokinetic coefficients of Clostridium cadaveris and Clostridium sporogenes, which utilize protein as carbon source. Experimental data on changes in peptone concentration and 16S rRNA gene copy numbers of C. cadaveris and C. sporogenes were fitted to model. The fourth-order Runge-Kutta approximation with non-linear least squares analysis was employed to solve the ordinary differential equations to estimate biokinetic coefficients. The maximum specific growth rate (μmax), half-saturation concentration (Ks), growth yield (Y), and decay coefficient (Kd) of C. cadaveris and C.sporogenes were 0.73 ± 0.05 and 1.35 ± 0.32 h-1, 6.07 ± 1.52 and 5.67 ± 1.53 g/l, 2.25 ± 0.75 x 1010 and 7.92 ± 3.71 x 109 copies/g, 0.002 ± 0.003 and 0.002 ± 0.001 h-1, respectively. The theoretical specific growth rate of C. sporogenes always exceeded that of C. cadaveris at peptone concentration higher than 3.62 g/l. When the influent peptone concentration was 5.0 g/l, the concentration of C.cadaveris gradually decreased to the steady value of 2.9 x 1010 copies/ml at 4 h Hydraulic retention time (HRT), which indicates a 67.1% reduction of the initial population, but the wash out occurred at HRTs of 1.9 and 3.2 h. The 16S rRNA gene copy numbers of C. sporogenes gradually decreased to steady values ranging from 1.1 x 1010 to 2.9 x 1010 copies/ml. C. sporogenes species was predicted to wash out at an HRT of 1.6 h.

9Antifungal Activity of Silver Salts of Keggin-Type Heteropolyacids Against Sporothrix spp.

저자 : Luciana Da Silva Mathias , João Carlos De Aquino Almeida , Luis César Passoni , Cristiani Miranda David Gossani , Gabriel Bonan Taveira , Valdirene Moreira Gomes , Olney Vieira-da-motta

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 540-551 (12 pages)

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Sporotrichosis is a chronic and subacute mycosis causing epidemiological outbreaks involving sick cats and humans in southeastern Brazil. The systemic disease prevails in cats and in humans, with the symptoms restricted to the skin of immunocompetent individuals. Under these conditions, the prolonged treatment of animals and cases of recurrence justify the discovery of new treatments for sporotrichosis. This work addresses the antifungal activity of silver salts of Keggin-type heteropolyacid salts (Ag-HPA salts) such as Ag3[PW12O40], Ag6[SiW10V2O40], Ag4[SiW12O40] and Ag3[PMo12O40] and interactions with the antifungal drugs itraconazole (ITC), terbinafine (TBF) and amphotericin B (AMB) on the yeast and mycelia forms of Sporothrix spp. Sporothrix spp. yeast cells were susceptible to Ag-HPA salts at minimum inhibitory concentration (MIC) values ranging from 8 to 128 μg/ml. Interactions between Ag3[PW12O40] and Ag3[PMo12O40] with itraconazole and amphotericin B resulted in higher antifungal activity with a reduction in growth and melanization. Treated cells showed changes in cell membrane integrity, vacuolization, cytoplasm disorder, and membrane detachment. Promising antifungal activity for treating sporotrichosis was observed for the Ag-HPA salts Ag3[PMo12O40] and Ag3[PW12O40], which have a low cost, high yield and activity at low concentrations. However, further evaluation of in vivo tests is still required.

10Two Flavonoid-Based Compounds from Murraya paniculata as Novel Human Carbonic Anhydrase Isozyme II Inhibitors Detected by a Resazurin Yeast-Based Assay

저자 : Anyaporn Sangkaew , Nawara Samritsakulchai , Kamonpan Sanachai , Thanyada Rungrotmongkol , Warinthorn Chavasiri , Chulee Yompakdee

발행기관 : 한국미생물생명공학회(구 한국산업미생물학회) 간행물 : Journal of Microbiology and Biotechnology 30권 4호 발행 연도 : 2020 페이지 : pp. 552-560 (9 pages)

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Human carbonic anhydrase (CA) isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3', 4', 5'-heptamethoxyflavone (4) and 3 ,5, 7, 8, 3', 4', 5'-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 μM, respectively, while they both exhibited no cytotoxic effect, even at the highest concentration tested (170 μM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC50 values of 24.0 and 34.3 μM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and SwissDock software. This revealed that compound 4 coordinated with the Zn2+ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 A (FlexX) or 2.29 A (SwissDock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.

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Mycobiology
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