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한국미생물생명공학회> Journal of Microbiology and Biotechnology> Genomic Analysis of Halotolerant Bacterial Strains Martelella soudanensis NC18T and NC20

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Genomic Analysis of Halotolerant Bacterial Strains Martelella soudanensis NC18T and NC20

Jung-yun Lee , Dong-hun Kim
  • : 한국미생물생명공학회
  • : Journal of Microbiology and Biotechnology 32권11호
  • : 연속간행물
  • : 2022년 11월
  • : 1427-1434(8pages)
Journal of Microbiology and Biotechnology

DOI


목차

Introduction
Materials and Methods
Results and Discussion
Acknowledgments
Conflicts of Interest
References

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Two novel, halotolerant strains of Martelella soudanensis, NC18T and NC20, were isolated from deep subsurface sediment, deeply sequenced, and comparatively analyzed with related strains. Based on a phylogenetic analysis using 16S rRNA gene sequences, the two strains grouped with members of the genus Martelella. Here, we sequenced the complete genomes of NC18T and NC20 to understand the mechanisms of their halotolerance. The genome sizes and G+C content of the strains were 6.1 Mb and 61.8 mol%, respectively. Moreover, NC18T and NC20 were predicted to contain 5,849 and 5,830 genes, and 5,502 and 5,585 protein-coding genes, respectively. Both strains contain the identically predicted 6 rRNAs and 48 tRNAs. The harboring of halotolerant-associated genes revealed that strains NC18T and NC20 might tolerate high salinity through the accumulation of potassium ions in a “salt-in” strategy induced by K+ uptake protein (kup) and the K+ transport system (trkAH and kdpFABC). These two strains also use the ectoine transport system (dctPQM), the glycine betaine transport system (proVWX), and glycine betaine uptake protein (opu) to accumulate “compatible solutes,” such as ectoine and glycine betaine, to protect cells from salt stress. This study reveals the halotolerance mechanism of strains NC18T and NC20 in high salt environments and suggests potential applications for these halotolerant and halophilic strains in environmental biotechnology.

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간행물정보

  • : 자연과학분야  > 생물
  • : KCI등재
  • : SCOPUS
  • : 월간
  • : 1017-7825
  • : 1738-8872
  • : 학술지
  • : 연속간행물
  • : 1991-2022
  • : 5943


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1Personalized Diets based on the Gut Microbiome as a Target for Health Maintenance: from Current Evidence to Future Possibilities

저자 : Eun-ji Song , Ji-hee Shin

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1497-1505 (9 pages)

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Recently, the concept of personalized nutrition has been developed, which states that food components do not always lead to the same metabolic responses, but vary from person to person. Although this concept has been studied based on individual genetic backgrounds, researchers have recently explored its potential role in the gut microbiome. The gut microbiota physiologically communicates with humans by forming a bidirectional relationship with the micronutrients, macronutrients, and phytochemicals consumed by the host. Furthermore, the gut microbiota can vary from person to person and can be easily shifted by diet. Therefore, several recent studies have reported the application of personalized nutrition to intestinal microflora. This review provides an overview of the interaction of diet with the gut microbiome and the latest evidence in understanding the inter-individual differences in dietary responsiveness according to individual baseline gut microbiota and microbiome-associated dietary intervention in diseases. The diversity of the gut microbiota and the presence of specific microorganisms can be attributed to physiological differences following dietary intervention. The difference in individual responsiveness based on the gut microbiota has the potential to become an important research approach for personalized nutrition and health management, although further well-designed large-scale studies are warranted.

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2Functional Roles of Exosomes in Allergic Contact Dermatitis

저자 : Bocui Song , Qian Chen , Yuqi Li , Shuang Zhan , Rui Zhao , Xue Shen , Min Liu , Chunyu Tong

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1506-1514 (9 pages)

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Allergic contact dermatitis (ACD) is an allergen-specific T-cell-mediated inflammatory response, albeit with unclear pathogenesis. Exosomes are nanoscale extracellular vesicles secreted by several cell types and widely distributed in various biological fluids. Exosomes affect the occurrence and development of ACD through immunoregulation among other ways. Nevertheless, the role of exosomes in ACD warrants further clarification. This review examines the progress of research into exosomes and their involvement in the pathogenesis, diagnosis, and treatment of ACD and provides ideas for exploring new diagnostic and treatment methods for this disease.

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3Dynamics of Viral and Host 3D Genome Structure upon Infection

저자 : Meyer J. Friedman , Haram Lee , Young-chan Kwon , Soohwan Oh

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1515-1526 (12 pages)

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Eukaryotic chromatin is highly organized in the 3D nuclear space and dynamically regulated in response to environmental stimuli. This genomic organization is arranged in a hierarchical fashion to support various cellular functions, including transcriptional regulation of gene expression. Like other host cellular mechanisms, viral pathogens utilize and modulate host chromatin architecture and its regulatory machinery to control features of their life cycle, such as lytic versus latent status. Combined with previous research focusing on individual loci, recent global genomic studies employing conformational assays coupled with high-throughput sequencing technology have informed models for host and, in some cases, viral 3D chromosomal structure re-organization during infection and the contribution of these alterations to virus-mediated diseases. Here, we review recent discoveries and progress in host and viral chromatin structural dynamics during infection, focusing on a subset of DNA (human herpesviruses and HPV) as well as RNA (HIV, influenza virus and SARS-CoV-2) viruses. An understanding of how host and viral genomic structure affect gene expression in both contexts and ultimately viral pathogenesis can facilitate the development of novel therapeutic strategies.

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4The ybcF Gene of Escherichia coli Encodes a Local Orphan Enzyme, Catabolic Carbamate Kinase

저자 : Nam Yeun Kim , Ok Bin Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1527-1536 (10 pages)

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Escherichia coli can use allantoin as its sole nitrogen source under anaerobic conditions. The ureidoglycolate produced by double release of ammonia from allantoin can flow into either the glyoxylate shunt or further catabolic transcarbamoylation. Although the former pathway is well studied, the genes of the latter (catabolic) pathway are not known. In the catabolic pathway, ureidoglycolate is finally converted to carbamoyl phosphate (CP) and oxamate, and then CP is dephosphorylated to carbamate by a catabolic carbamate kinase (CK), whereby ATP is formed. We identified the ybcF gene in a gene cluster containing fdrA-ylbE-ylbF-ybcF that is located downstream of the allDCE-operon. Reverse transcription PCR of total mRNA confirmed that the genes fdrA, ylbE, ylbF, and ybcF are co-transcribed. Deletion of ybcF caused only a slight increase in metabolic flow into the glyoxylate pathway, probably because CP was used to de novo synthesize pyrimidine and arginine. The activity of the catabolic CK was analyzed using purified YbcF protein. The Vmax is 1.82 U/mg YbcF for CP and 1.94 U/mg YbcF for ADP, and the KM value is 0.47 mM for CP and 0.43 mM for ADP. With these results, it was experimentally revealed that the ybcF gene of E. coli encodes catabolic CK, which completes anaerobic allantoin degradation through substrate-level phosphorylation. Therefore, we suggest renaming the ybcF gene as allK.

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5Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus from Nasal Specimens: Overcoming MRSA with Silver Nanoparticles and Their Applications

저자 : Aly E. Abo-amer , Sanaa M. F. Gad El-rab , Eman M. Halawani , Ameen M. Niaz , Mohammed S. Bamaga

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1537-1546 (10 pages)

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Staphylococcus aureus is a cause of high mortality in humans and therefore it is necessary to prevent its transmission and reduce infections. Our goals in this research were to investigate the frequency of methicillin-resistant S. aureus (MRSA) in Taif, Saudi Arabia, and assess the relationship between the phenotypic antimicrobial sensitivity patterns and the genes responsible for resistance. In addition, we examined the antimicrobial efficiency and application of silver nanoparticles (AgNPs) against MRSA isolates. Seventy-two nasal swabs were taken from patients; MRSA was cultivated on Mannitol Salt Agar supplemented with methicillin, and 16S rRNA sequencing was conducted in addition to morphological and biochemical identification. Specific resistance genes such as ermAC, aacA-aphD, tetKM, vatABC and mecA were PCR-amplified and resistance plasmids were also investigated. The MRSA incidence was ~49 % among the 72 S. aureus isolates and all MRSA strains were resistant to oxacillin, penicillin, and cefoxitin. However, vancomycin, linezolid, teicoplanin, mupirocin, and rifampicin were effective against 100% of MRSA strains. About 61% of MRSA strains exhibited multidrug resistance and were resistant to 3-12 antimicrobial medications (MDR). Methicillin resistance gene mecA was presented in all MDR-MRSA strains. Most MDR-MRSA contained a plasmid of > 10 kb. To overcome bacterial resistance, AgNPs were applied and displayed high antimicrobial activity and synergistic effect with penicillin. Our findings may help establish programs to control bacterial spread in communities as AgNPs appeared to exert a synergistic effect with penicillin to control bacterial resistance.

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6Antibacterial Mode of Action of β-Amyrin Promotes Apoptosis-Like Death in Escherichia coli by Producing Reactive Oxygen Species

저자 : Giyeol Han , Dong Gun Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1547-1552 (6 pages)

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β-Amyrin is a pentacyclic triterpene widely distributed in leaves and stems worldwide. The ability of β-amyrin to induce the production of reactive oxygen species (ROS) in microorganisms suggests its potential as an antimicrobial agent. Thus, this study aimed to elucidate the antibacterial mode of action of β-amyrin. We treated Escherichia coli cells with β-amyrin and found that it triggered ROS accumulation. Excessive stress caused by ROS, particularly hydroxyl radicals, induces glutathione (GSH) dysfunction. GSH protects cells from oxidative and osmotic stresses; thus, its dysfunction leads to membrane depolarization. The resultant change in membrane potential leads to the release of apoptotic proteins, such as caspases. The activated caspases-like protein promotes the cleavage of DNA into single strands, which is a hallmark of apoptosis-like death in bacteria. Apoptotic cells usually undergo events such as DNA fragmentation and phosphatidylserine exposure, differentiating them from necrotic cells, and the cells treated with β-amyrin in this study were positive for annexin V and negative for propidium iodide, indicating apoptosis-like death. In conclusion, our findings suggest that the antibacterial mode of action of β-amyrin involves the induction of ROS, which resulted in apoptosis-like death in E. coli.

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7Mucilaginibacter aquariorum sp. nov., Isolated from Fresh Water

저자 : Ve Van Le , So-ra Ko , Mingyeong Kang , Hee-mock Oh , Chi-yong Ahn

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1553-1560 (8 pages)

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A Gram-stain-negative, rod-shaped bacterial strain, JC4T, was isolated from a freshwater sample and determined the taxonomic position. Initial identification based on 16S rRNA gene sequences revealed that strain JC4T is affiliated to the genus Mucilaginibacter with a sequence similarity of 97.97% to Mucilaginibacter rigui WPCB133T. The average nucleotide identity and digital DNA-DNA hybridization values between strain JC4T and Mucilaginibacter species were estimated below 80.92% and 23.9%, respectively. Strain JC4T contained summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and iso-C15:0 as predominant cellular fatty acids. The dominant polar lipids were identified as phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid, and two unidentified lipids. The respiratory quinone was MK-7. The genomic DNA G+C content of strain JC4T was determined to be 42.44%. The above polyphasic evidences support that strain JC4T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter aquariorum sp. nov. is proposed. The type strain is JC4T (= KCTC 92230T = LMG 32715T).

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8Distinct Bacterial and Fungal Communities Colonizing Waste Plastic Films Buried for More Than 20 Years in Four Landfill Sites in Korea

저자 : Joon-hui Chung , Jehyeong Yeon , Hoon Je Seong , Si-hyun An , Da-yeon Kim , Younggun Yoon , Hang-yeon Weon , Jeong Jun Kim , Jae-hyung Ahn

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1561-1572 (12 pages)

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Plastic pollution has been recognized as a serious environmental problem, and microbial degradation of plastics is a potential, environmentally friendly solution to this. Here, we analyzed and compared microbial communities on waste plastic films (WPFs) buried for long periods at four landfill sites with those in nearby soils to identify microbes with the potential to degrade plastics. Fourier-transform infrared spectroscopy spectra of these WPFs showed that most were polyethylene and had signs of oxidation, such as carbon-carbon double bonds, carbon-oxygen single bonds, or hydrogen-oxygen single bonds, but the presence of carbonyl groups was rare. The species richness and diversity of the bacterial and fungal communities on the films were generally lower than those in nearby soils. Principal coordinate analysis of the bacterial and fungal communities showed that their overall structures were determined by their geographical locations; however, the microbial communities on the films were generally different from those in the soils. For the pulled data from the four landfill sites, the relative abundances of Bradyrhizobiaceae, Pseudarthrobacter, Myxococcales, Sphingomonas, and Spartobacteria were higher on films than in soils at the bacterial genus level. At the species level, operational taxonomic units classified as Bradyrhizobiaceae and Pseudarthrobacter in bacteria and Mortierella in fungi were enriched on the films. PICRUSt analysis showed that the predicted functions related to amino acid and carbohydrate metabolism and xenobiotic degradation were more abundant on films than in soils. These results suggest that specific microbial groups were enriched on the WPFs and may be involved in plastic degradation.

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9Printing Optimization of 3D Structure with Lard-like Texture Using a Beeswax-Based Oleogels

저자 : Hyeona Kang , Yourim Oh , Nam Keun Lee , Jin-kyu Rhee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1573-1582 (10 pages)

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In this study, we investigated the optimal conditions for 3D structure printing of alternative fats that have the textural properties of lard using beeswax (BW)-based oleogel by a statistical analysis. Products printed with over 15% BW oleogel at 50% and 75% infill level (IL) showed high printing accuracy with the lowest dimensional printing deviation for the designed model. The hardness, cohesion, and adhesion of printed samples were influenced by BW concentration and infill level. For multi-response optimization, fixed target values (hardness, adhesiveness, and cohesiveness) were applied with lard printed at 75% IL. The preparation parameters obtained as a result of multiple reaction prediction were 58.9% IL and 16.0% BW, and printing with this oleogel achieved fixed target values similar to those of lard. In conclusion, our study shows that 3D printing based on the BW oleogel system produces complex internal structures that allow adjustment of the textural properties of the printed samples, and BW oleogels could potentially serve as an excellent replacement for fat.

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10Lactic Acid Bacteria Strains Used as Starters for Kimchi Fermentation Protect the Disruption of Tight Junctions in the Caco-2 Cell Monolayer Model

저자 : Jin Yong Kang , Moeun Lee , Jung Hee Song , Eun Ji Choi , Da Un Kim , Seul Ki Lim , Namhee Kim , Ji Yoon Chang

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 12호 발행 연도 : 2022 페이지 : pp. 1583-1588 (6 pages)

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In this study, we investigated the effect of lactic acid bacteria (LAB) strains used as starters for kimchi fermentation, namely Lactococcus lactis WiKim0124, Companilactobacillus allii WiKim39, Leuconostoc mesenteroides WiKim0121Leuconostoc mesenteroides WiKim33, and Leuconostoc mesenteroides WiKim32, on the intestinal epithelial tight junctions (TJs). These LAB strains were not cytotoxic to Caco-2 cells at 500 μg/ml concentration. In addition, hydrogen peroxide (H2O2) decreased Caco-2 viability, but the LAB strains protected the cells against H2O2-induced cytotoxicity. We also found that lipopolysaccharide (LPS) promoted Caco-2 proliferation; however, no specific changes were observed upon treatment with LAB strains and LPS. Our evaluation of the permeability in the Caco-2 monolayer model confirmed its increase by both LPS and H2O2. The LAB strains inhibited the increase in permeability by protecting TJs, which we evaluated by measuring TJ proteins such as zonula occludens-1 and occludin, and analyzing them by western blotting and immunofluorescence staining. Our findings show that LAB strains used for kimchi fermentation can suppress the increase in intestinal permeability due to LPS and H2O2 by protecting TJs. Therefore, these results suggest the possibility of enhancing the functionality of kimchi through its fermentation using functional LAB strains.

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1Biotechnological Approaches for Biomass and Lipid Production Using Microalgae Chlorella and Its Future Perspectives

저자 : Sujeong Je , Yasuyo Yamaoka

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1357-1372 (16 pages)

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Heavy reliance on fossil fuels has been associated with increased climate disasters. As an alternative, microalgae have been proposed as an effective agent for biomass production. Several advantages of microalgae include faster growth, usage of non-arable land, recovery of nutrients from wastewater, efficient CO2 capture, and high amount of biomolecules that are valuable for humans. Microalgae Chlorella spp. are a large group of eukaryotic, photosynthetic, unicellular microorganisms with high adaptability to environmental variations. Over the past decades, Chlorella has been used for the large-scale production of biomass. In addition, Chlorella has been actively used in various food industries for improving human health because of its antioxidant, antidiabetic, and immunomodulatory functions. However, the major restrictions in microalgal biofuel technology are the cost-consuming cultivation, processing, and lipid extraction processes. Therefore, various trials have been performed to enhance the biomass productivity and the lipid contents of Chlorella cells. This study provides a comprehensive review of lipid enhancement strategies mainly published in the last five years and aimed at regulating carbon sources, nutrients, stresses, and expression of exogenous genes to improve biomass production and lipid synthesis.

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2Fructan Biosynthesis by Yeast Cell Factories

저자 : Hyunjun Ko , Bong Hyun Sung , Mi-jin Kim , Jung-hoon Sohn , Jung-hoon Bae

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1373-1381 (9 pages)

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Fructan is a polysaccharide composed of fructose and can be classified into several types, such as inulin, levan, and fructo-oligosaccharides, based on their linkage patterns and degree of polymerization. Owing to its structural and functional diversity, fructan has been used in various fields including prebiotics, foods and beverages, cosmetics, and pharmaceutical applications. With increasing interest in fructans, efficient and straightforward production methods have been explored. Since the 1990s, yeast cells have been employed as producers of recombinant enzymes for enzymatic conversion of fructans including fructosyltransferases derived from various microbes and plants. More recently, yeast cell factories are highlighted as efficient workhorses for fructan production by direct fermentation. In this review, recent advances and strategies for fructan biosynthesis by yeast cell factories are discussed.

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3Asterias pectinifera-Derived Collagen Peptides Mixed with Halocynthia roretzi Extracts Exhibit Anti-Photoaging Activities during Exposure to UV Irradiation, and Antibacterial Properties

저자 : Soo-jin Oh , Ji-ye Park , Bada Won , Yong-taek Oh , Seung-chan Yang , Ok Sarah Shin

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1382-1389 (8 pages)

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Asterias pectinifera, a species of starfish and cause of concern in the aquaculture industry, was recently identified as a source of non-toxic and highly water-soluble collagen peptides. In this study, we investigated the antioxidant and anti-photoaging functions of compounds formulated using collagen peptides from extracts of Asterias pectinifera and Halocynthia roretzi (AH). Our results showed that AH compounds have various skin protective functions, including antioxidant effects, determined by measuring the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl radicals, as well as anti-melanogenic effects, determined by measuring tyrosinase inhibition activity. To determine whether ethosome-encapsulated AH compounds (E(AH)) exert ultraviolet (UV)-protective effects, human dermal fibroblasts or keratinocytes were incubated with E(AH) before and after exposure to UVA or UVB. E(AH) treatment led to inhibition of photoaging-induced secretion of matrix metalloproteinase-1 and interleukin-6 and -8, which are associated with inflammatory responses during UV irradiation. Finally, the antibacterial effects of AH and E(AH) were confirmed against both gram-negative and gram-positive bacteria. Our results indicate that E(AH) has the potential for use in the development of cosmetics with a range of skin protective functions.

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4Sulforaphene Attenuates Cutibacterium acnes-Induced Inflammation

저자 : Hwan Ju Hwang , Jong-eun Kim , Ki Won Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1390-1395 (6 pages)

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Acne is a chronic inflammatory disease of the sebaceous gland attached to the hair follicles. Cutibacterium acnes is a major cause of inflammation caused by acne. It is well known that C. acnes secretes a lipolytic enzyme to break down lipids in sebum, and free fatty acids produced at this time accelerate the inflammatory reaction. There are several drugs used to treat acne; however, each one has various side effects. According to previous studies, sulforaphene (SFEN) has several functions associated with lipid metabolism, brain function, and antibacterial and anti-inflammatory activities. In this study, we examined the effects of SFEN on bacterial growth and inflammatory cytokine production induced by C. acnes. The results revealed that SFEN reduced the growth of C. acnes and inhibited proinflammatory cytokines in C. acnes-treated HaCaT keratinocytes through inhibiting NF-κB-related pathways. In addition, SFEN regulated the expression level of IL-1α, a representative pro-inflammatory cytokine expressed in co-cultured HaCaT keratinocytes and THP-1 monocytes induced by C. acnes. In conclusion, SFEN showed antibacterial activity against C. acnes and controlled the inflammatory response on keratinocytes and monocytes. This finding means that SFEN has potential as both a cosmetic material for acne prevention and a pharmaceutical material for acne treatment.

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5Inactivated Vibrio cholerae Strains That Express TcpA via the toxT-139F Allele Induce Antibody Responses against TcpA

저자 : Eun Jin Kim , Jonghyun Bae , Young-jun Ju , Do-bin Ju , Donghyun Lee , Seonghyeon Son , Hunseok Choi , Thandavarayan Ramamurthy , Cheol-heui Yun , Dong Wook Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1396-1405 (10 pages)

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Cholera remains a major global public health problem, for which oral cholera vaccines (OCVs) being a valuable strategy. Patients, who have recovered from cholera, develop antibody responses against LPS, cholera toxin (CT), toxin-coregulated pilus (TCP) major subunit A (TcpA) and other antigens; thus, these responses are potentially important contributors to immunity against Vibrio cholerae infection. However, assessments of the efficacy of current OCVs, especially inactivated OCVs, have focused primarily on O-antigen-specific antibody responses, suggesting that more sophisticated strategies are required for inactivated OCVs to induce immune responses against TCP, CT, and other antigens. Previously, we have shown that the toxT-139F allele enables V. cholerae strains to produce CT and TCP under simple laboratory culture conditions. Thus, we hypothesized that V. cholerae strains that express TCP via the toxT-139F allele induce TCP-specific antibody responses. As anticipated, V. cholerae strains that expressed TCP through the toxT-139F allele elicited antibody responses against TCP when the inactivated bacteria were delivered via a mouse model. We have further developed TCP-expressing V. cholerae strains that have been used in inactivated OCVs and shown that they effect an antibody response against TcpA in vivo, suggesting that V. cholerae strains with the toxT-139F allele are excellent candidates for cholera vaccines.

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6Silencing YY1 Alleviates Ox-LDL-Induced Inflammation and Lipid Accumulation in Macrophages through Regulation of PCSK9/ LDLR Signaling

저자 : Zhengyao Qian , Jianping Zhao

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1406-1415 (10 pages)

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The formation of macrophage foam cells stimulated by oxidized low-density lipoprotein (ox-LDL) is deemed an important cause of atherosclerosis. Transcription factor Yin Yang 1 (YY1), which is a universally expressed multifunctional protein, is closely related to cell metabolism disorders such as lipid metabolism, sugar metabolism, and bile acid metabolism. However, whether YY1 is involved in macrophage inflammation and lipid accumulation still remains unknown. After mouse macrophage cell line RAW264.7 cells were induced by ox-LDL, YY1 and proprotein convertase subtilisin/kexin type 9 (PCSK9) expressions were found to be increased while low-density lipoprotein receptor (LDLR) expression was lowly expressed. Subsequently, through reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot analysis, Oil Red O staining and cholesterol quantification, it turned out that silencing of YY1 attenuated the inflammatory response and lipid accumulation in RAW264.7 cells caused by ox-LDL. Moreover, results from the JASPAR database, chromatin immunoprecipitation (ChIP) assay, luciferase reporter assay and Western blot analysis suggested that YY1 activated PCSK9 by binding to PCSK9 promoter and modulated the expression of LDLR in the downstream of PCSK9. In addition, the results of functional experiments demonstrated that the inhibitory effects of YY1 interference on ox-LDL-mediated macrophage inflammation and lipid accumulation were reversed by PCSK9 overexpression. To sum up, YY1 depletion inhibited its activation of PCSK9, thereby reducing cellular inflammatory response, cholesterol homeostasis imbalance, and lipid accumulation caused by ox-LDL.

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7Characterization of Pseudomonas sp. NIBR-H-19, an Antimicrobial Secondary Metabolite Producer Isolated from the Gut of Korean Native Sea Roach, Ligia exotica

저자 : Sungmin Hwang , Jun Hyeok Yang , Ho Seok Sim , Sung Ho Choi , Byounghee Lee , Woo Young Bang , Ki Hwan Moon

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1416-1426 (11 pages)

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The need to discover new types of antimicrobial agents has grown since the emergence of antibioticresistant pathogens that threaten human health. The world's oceans, comprising complex niches of biodiversity, are a promising environment from which to extract new antibiotics-like compounds. In this study, we newly isolated Pseudomonas sp. NIBR-H-19 from the gut of the sea roach Ligia exotica and present both phenotypes and genomic information consisting of 6,184,379 bp in a single chromosome possessing a total of 5,644 protein-coding genes. Genomic analysis of the isolated species revealed that numerous genes involved in antimicrobial secondary metabolites are predicted throughout the whole genome. Moreover, our analysis showed that among twenty-five pathogenic bacteria, the growth of three pathogens, including Staphylococcus aureus, Streptococcus hominis and Rhodococcus equi, was significantly inhibited by the culture of Pseudomonas sp. NIBR-H- 19. The characterization of marine microorganisms with biochemical assays and genomics tools will help uncover the biosynthesis and action mechanism of antimicrobial metabolites for development as antagonistic probiotics against fish pathogens in an aquatic culture system.

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8Genomic Analysis of Halotolerant Bacterial Strains Martelella soudanensis NC18T and NC20

저자 : Jung-yun Lee , Dong-hun Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1427-1434 (8 pages)

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Two novel, halotolerant strains of Martelella soudanensis, NC18T and NC20, were isolated from deep subsurface sediment, deeply sequenced, and comparatively analyzed with related strains. Based on a phylogenetic analysis using 16S rRNA gene sequences, the two strains grouped with members of the genus Martelella. Here, we sequenced the complete genomes of NC18T and NC20 to understand the mechanisms of their halotolerance. The genome sizes and G+C content of the strains were 6.1 Mb and 61.8 mol%, respectively. Moreover, NC18T and NC20 were predicted to contain 5,849 and 5,830 genes, and 5,502 and 5,585 protein-coding genes, respectively. Both strains contain the identically predicted 6 rRNAs and 48 tRNAs. The harboring of halotolerant-associated genes revealed that strains NC18T and NC20 might tolerate high salinity through the accumulation of potassium ions in a “salt-in” strategy induced by K+ uptake protein (kup) and the K+ transport system (trkAH and kdpFABC). These two strains also use the ectoine transport system (dctPQM), the glycine betaine transport system (proVWX), and glycine betaine uptake protein (opu) to accumulate “compatible solutes,” such as ectoine and glycine betaine, to protect cells from salt stress. This study reveals the halotolerance mechanism of strains NC18T and NC20 in high salt environments and suggests potential applications for these halotolerant and halophilic strains in environmental biotechnology.

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9Zinc-Solubilizing Streptomyces spp. as Bioinoculants for Promoting the Growth of Soybean (Glycine max (L.) Merrill)

저자 : Chanwit Suriyachadkun , Orawan Chunhachart , Moltira Srithaworn , Rungnapa Tangchitcharoenkhul , Janpen Tangjitjareonkun

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1435-1446 (12 pages)

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Zinc-solubilizing bacteria can convert the insoluble form of zinc into soluble forms available to plants. This study was conducted to isolate and screen zinc-solubilizing actinobacteria from rhizosphere soils and to assess their effect on vegetable soybean growth. In total, 200 actinobacteria strains belonging to 10 genera were isolated from rhizosphere soil samples. Among these isolates, four showed zinc solubilization with solubilizing index values ranging from 3.11 to 3.78 on Bunt and Rovira agar supplemented with 0.1% zinc oxide. For the quantitative assay, in broth culture, strains CME34 and EX51 solubilized maximum available zinc contents of 529.71 and 243.58 μg/ml. Furthermore, indole-3-acetic acid (IAA) and ammonia were produced by these two strains, the strain CME34 produced the highest amount of IAA 4.62 μg/ml and the strain EX51 produced the highest amount of ammonia 361.04 μg/ml. In addition, the phosphate-solubilizing abilities in Pikovskaya's medium of CME34 and EX51 were 64.67 and 115.67 μg/ml. Based on morphological and biochemical characterization and 16S rDNA sequencing, the strains CME34 and EX51 were closely related to the genus Streptomyces. In a greenhouse experiment, single-strain inoculation of Streptomyces sp. CME34 or EX51 significantly increased the shoot length, root length, plant dry weight, number of pods per plant and number of seeds per plant of vegetable soybean plants compared to the uninoculated control. These findings facilitated the conclusion that the two Streptomyces strains have potential as zinc solubilizers and can be suggested as bioinoculants to promote the growth and yield of soybean.

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10The Relationship between Prohibitin 1 Expression, Hepatotoxicity Induced by Acetaminophen, and Hepatoprotection by S-Adenosylmethionine in AML12 Cells

저자 : Eunhye Cho , Soohan Jung , Jina Kim , Kwang Suk Ko

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 11호 발행 연도 : 2022 페이지 : pp. 1447-1453 (7 pages)

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Prohibitin 1 (Phb1) is a pleiotropic protein, located mainly in the mitochondrial inner membrane and involved in the regulation of cell proliferation and the stabilization of mitochondrial protein. Acetaminophen (APAP) is one of the most commonly used over-the-counter analgesics worldwide. However, at high dose, the accumulation of N-acetyl-p-benzoquinone imine (NAPQI) can lead to APAP-induced hepatotoxicity. In this study, we sought to understand the regulation of mRNA expression in relation to APAP and GSH metabolism by Phb1 in normal mouse AML12 hepatocytes. We used two different Phb1 silencing levels: high-efficiency (HE, >90%) and low-efficiency (LE, 50- 60%). In addition, the siRNA-transfected cells were further pretreated with 0.5 mM of Sadenosylmethionine (SAMe) for 24 h before treatment with APAP at different doses (1-2 mM) for 24 h. The expression of APAP metabolism-related and antioxidant genes such as Cyp2e1 and Ugt1a1 were increased during SAMe pretreatment. Moreover, SAMe increased intracellular GSH concentration and it was maintained after APAP treatment. To sum up, Phb1 silencing and APAP treatment impaired the metabolism of APAP in hepatocytes, and SAMe exerted a protective effect against hepatotoxicity by upregulating antioxidant genes.

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