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한국미생물생명공학회> Journal of Microbiology and Biotechnology> Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach

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Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach

Juyoun Shin , Sun Shin , Seung-hyun Jung , Chulmin Park , Sung-yeon Cho , Dong-gun Lee , Yeun-jun Chung
  • : 한국미생물생명공학회
  • : Journal of Microbiology and Biotechnology 31권11호
  • : 연속간행물
  • : 2021년 11월
  • : 1481-1489(9pages)
Journal of Microbiology and Biotechnology

DOI


목차

Introduction
Materials and Methods
Results
Discussion
Acknowledgments
Conflicts of Interests
References

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초록 보기

Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.

UCI(KEPA)

I410-ECN-0102-2022-400-000829098

간행물정보

  • : 자연과학분야  > 생물
  • : KCI등재
  • : SCOPUS
  • : 월간
  • : 1017-7825
  • : 1738-8872
  • : 학술지
  • : 연속간행물
  • : 1991-2022
  • : 5804


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1Strategies to Mitigate Enteric Methane Emissions from Ruminant Animals

저자 : Tenzin Tseten , Rey Anthony Sanjorjo , Moonhyuk Kwon , Seon-won Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 269-277 (9 pages)

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Human activities account for approximately two-thirds of global methane emissions, wherein the livestock sector is the single massive methane emitter. Methane is a potent greenhouse gas of over 21 times the warming effect of carbon dioxide. In the rumen, methanogens produce methane as a by-product of anaerobic fermentation. Methane released from ruminants is considered as a loss of feed energy that could otherwise be used for productivity. Economic progress and growing population will inflate meat and milk product demands, causing elevated methane emissions from this sector. In this review, diverse approaches from feed manipulation to the supplementation of organic and inorganic feed additives and direct-fed microbial in mitigating enteric methane emissions from ruminant livestock are summarized. These approaches directly or indirectly alter the rumen microbial structure thereby reducing rumen methanogenesis. Though many inorganic feed additives have remarkably reduced methane emissions from ruminants, their usage as feed additives remains unappealing because of health and safety concerns. Hence, feed additives sourced from biological materials such as direct-fed microbials have emerged as a promising technique in mitigating enteric methane emissions.

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2An Engineered Outer Membrane-Defective Escherichia coli Secreting Protective Antigens against Streptococcus suis via the Twin-Arginine Translocation Pathway as a Vaccine

저자 : Wenyu Li , Fan Yin , Zixuan Bu , Yuying Liu , Yongqing Zhang , Xiabing Chen , Shaowen Li , Lu Li , Rui Zhou , Qi Huang

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 278-286 (9 pages)

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Live bacterial vector vaccines are one of the most promising vaccine types and have the advantages of low cost, flexibility, and good safety. Meanwhile, protein secretion systems have been reported as useful tools to facilitate the release of heterologous antigen proteins from bacterial vectors. The twin-arginine translocation (Tat) system is an important protein export system that transports fully folded proteins in a signal peptide-dependent manner. In this study, we constructed a live vector vaccine using an engineered commensal Escherichia coli strain in which amiA and amiC genes were deleted, resulting in a leaky outer membrane that allows the release of periplasmic proteins to the extracellular environment. The protective antigen proteins SLY, enolase, and Sbp against Streptococcus suis were targeted to the Tat pathway by fusing a Tat signal peptide. Our results showed that by exploiting the Tat pathway and the outer membrane-defective E. coli strain, the antigen proteins were successfully secreted. The strains secreting the antigen proteins were used to vaccinate mice. After S. suis challenge, the vaccinated group showed significantly higher survival and milder clinical symptoms compared with the vector group. Further analysis showed that the mice in the vaccinated group had lower burdens of bacteria load and slighter pathological changes. Our study reports a novel live bacterial vector vaccine that uses the Tat system and provides a new alternative for developing S. suis vaccine.

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3Electron Transfer to Hydroxylase through Component Interactions in Soluble Methane Monooxygenase

저자 : Chaemin Lee , Yunha Hwang , Hyun Goo Kang , Seung Jae Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 287-293 (7 pages)

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The hydroxylation of methane (CH4) is crucial to the field of environmental microbiology, owing to the heat capacity of methane, which is much higher than that of carbon dioxide (CO2). Soluble methane monooxygenase (sMMO), a member of the bacterial multicomponent monooxygenase (BMM) superfamily, is essential for the hydroxylation of specific substrates, including hydroxylase (MMOH), regulatory component (MMOB), and reductase (MMOR). The diiron active site positioned in the MMOH α-subunit is reduced through the interaction of MMOR in the catalytic cycle. The electron transfer pathway, however, is not yet fully understood due to the absence of complex structures with reductases. A type II methanotroph, Methylosinus sporium 5, successfully expressed sMMO and hydroxylase, which were purified for the study of the mechanisms. Studies on the MMOH-MMOB interaction have demonstrated that Tyr76 and Trp78 induce hydrophobic interactions through π-π stacking. Structural analysis and sequencing of the ferredoxin domain in MMOR (MMOR-Fd) suggested that Tyr93 and Tyr95 could be key residues for electron transfer. Mutational studies of these residues have shown that the concentrations of flavin adenine dinucleotide (FAD) and iron ions are changed. The measurements of dissociation constants (Kds) between hydroxylase and mutated reductases confirmed that the binding affinities were not significantly changed, although the specific enzyme activities were significantly reduced by MMOR-Y93A. This result shows that Tyr93 could be a crucial residue for the electron transfer route at the interface between hydroxylase and reductase.

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4Appropriate Soil Heat Treatment Promotes Growth and Disease Suppression of Panax notoginseng by Interfering with the Bacterial Community

저자 : Ying-bin Li , Zhi-ping Zhang , Ye Yuan , Hui-chuan Huang , Xin-yue Mei , Fen Du , Min Yang , Yi-xiang Liu , Shu-sheng Zhu

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 294-301 (8 pages)

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In our greenhouse experiment, soil heat treatment groups (50, 80, and 121°C) significantly promoted growth and disease suppression of Panax notoginseng in consecutively cultivated soil (CCS) samples (p < 0.01), and 80°C worked better than 50°C and 121°C (p < 0.01). Furthermore, we found that heat treatment at 80°C changes the microbial diversity in CCS, and the inhibition ratios of culturable microorganisms, such as fungi and actinomycetes, were nearly 100%. However, the heat-tolerant bacterial community was preserved. The 16S rRNA gene and internal transcribed spacer (ITS) sequencing analyses indicated that the soil heat treatment had a greater effect on the Chao1 index and Shannon's diversity index of bacteria than fungi, and the relative abundances of Firmicutes and Proteobacteria were significantly higher than without heating (80 and 121°C, p < 0.05). Soil probiotic bacteria, such as Bacillus (67%), Sporosarcina (9%), Paenibacillus (6%), Paenisporosarcina (6%), and Cohnella (4%), remained in the soil after the 80°C and 121°C heat treatments. Although steam increased the relative abundances of most of the heat-tolerant microbes before sowing, richness and diversity gradually recovered to the level of CCS, regardless of fungi or bacteria, after replanting. Thus, we added heat-tolerant microbes (such as Bacillus) after steaming, which reduced the relative abundance of pathogens, recruited antagonistic bacteria, and provided a long-term protective effect compared to the steaming and Bacillus alone (p < 0.05). Taken together, the current study provides novel insight into sustainable agriculture in a consecutively cultivated system.

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5RK-270D and E, Oxindole Derivatives from Streptomyces sp. with Anti-Angiogenic Activity

저자 : Jun-pil Jang , Mina Jang , Toshihiko Nogawa , Shunji Takahashi , Hiroyuki Osada , Jong Seog Ahn , Sung-kyun Ko , Jae-hyuk Jang

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 302-306 (5 pages)

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A chemical investigation of a culture extract from Streptomyces sp. RK85-270 led to the isolation and characterization of two new oxindoles, RK-270D (1) and E (2). The structures of 1 and 2 were determined by analyzing spectroscopic and spectrometric data from 1D and 2D NMR and High-resolution electrospray ionization mass spectrometry (HRESIMS) experiments. Compound 1 exhibited anti-angiogenic activities against human umbilical vein endothelial cells (HUVECs) without cytotoxicity. Results of Western blot analysis revealed that 1 inhibits VEGF-induced angiogenesis in the HUVECs via VEGFR2/ p38 MAPK-mediated pathway.

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6Functional Characterization of khadi Yeasts Isolates for Selection of Starter Cultures

저자 : Koketso Motlhanka , Kebaneilwe Lebani , Mar Garcia-aloy , Nerve Zhou

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 307-316 (10 pages)

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Yeasts play an important role in spontaneous fermentation of traditional alcoholic beverages. Our previous study revealed that a mixed-consortia of both Saccharomyces and non-Saccharomyces yeasts were responsible for fermentation of khadi, a popular, non-standardized traditional beverage with an immense potential for commercialization in Botswana. Functional characterization of isolated fermenting yeasts from mixed consortia is an indispensable step towards the selection of potential starter cultures for commercialization of khadi. In this study, we report the characterization of 13 khadi isolates for the presence of brewing-relevant phenotypes such as their fermentative capacity, ability to utilize a range of carbon sources and their ability to withstand brewing-associated stresses, as a principal step towards selection of starter cultures. Khadi isolates such as Saccharomyces cerevisiae, Saccharomycodes ludwigii and Candida ethanolica showed good brewing credentials but Lachancea fermentati emerged as the isolate with the best brewing attributes with a potential as a starter culture. However, we were then prompted to investigate the potential of L. fermentati to influence the fruity aromatic flavor, characteristic of khadi. The aroma components of 18 khadi samples were extracted using headspace solid phase micro-extraction (HS-SPME) and identified using a GC-MS. We detected esters as the majority of volatile compounds in khadi, typical of the aromatic signature of both khadi and L. fermentati associated fermentations. This work shows that L. fermentati has potential for commercial production of khadi.

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7Comparison of Liquid and Solid-State Fermentation Processes for the Production of Enzymes and Beta-Glucan from Hulled Barley

저자 : Se Yeon Lee , Chae Hun Ra

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 317-323 (7 pages)

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Solid-state fermentation using hulled barley was carried out to produce enzymes and β-glucan. The one-factor-at-a-time experiments were carried out to determine the optimal composition of the basal medium. The modified synthetic medium composition in liquid-state fermentation was determined to be 70 g/l hulled barley, 0 g/l rice bran, 5 g/l soytone, and 6 g/l ascorbic acid. Optimal pretreatment conditions of hulled barley by solid-state fermentation were evaluated in terms of maximum production of fungal biomass, amylase, protease, and β-glucan, which were 1.26 mg/g, 31310.34 U/g, 2614.95 U/g, and 14.6% (w/w), respectively, at 60 min of pretreatment condition. Thus, the solid-state fermentation process was found to enhance the overall fermentation yields of hulled barley to produce high amounts of enzymes and β-glucan.

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8Heat-Killed Lactiplantibacillus plantarum LRCC5314 Mitigates the Effects of Stress-Related Type 2 Diabetes in Mice via Gut Microbiome Modulation

저자 : Yohan Nam , Seokmin Yoon , Jihye Baek , Jong-hwa Kim , Miri Park , Kwangwoo Hwang , Wonyong Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 324-332 (9 pages)

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The incidence of stress-related type 2 diabetes (stress-T2D), which is aggravated by physiological stress, is increasing annually. The effects of Lactobacillus, a key component of probiotics, have been widely studied in diabetes; however, studies on the effects of postbiotics are still limited. Here, we aimed to examine the mechanism through which heat-killed Lactiplantibacillus plantarum LRCC5314 (HK-LRCC5314) alleviates stress-T2D in a cold-induced stress-T2D C57BL/6 mouse model. HK-LRCC5314 markedly decreased body weight gain, adipose tissue (neck, subcutaneous, and epididymal) weight, and fasting glucose levels. In the adipose tissue, mRNA expression levels of stress-T2D associated factors (NPY, Y2R, GLUT4, adiponectin, and leptin) and pro-inflammatory factors (TNF-α, IL-6, and CCL-2) were also altered. Furthermore, HK-LRCC5314 increased the abundance of Barnesiella, Alistipes, and butyrate-producing bacteria, including Akkermansia, in feces and decreased the abundance of Ruminococcus, Dorea, and Clostridium. Thus, these findings suggest that HK-LRCC5314 exerts protective effects against stress-T2D via gut microbiome modulation, suggesting its potential as a supplement for managing stress-T2D.

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9Characterization of Prophages in Leuconostoc Derived from Kimchi and Genomic Analysis of the Induced Prophage in Leuconostoc lactis

저자 : Song-hee Kim , Jong-hyun Park

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 333-340 (8 pages)

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Leuconostoc has been used as a principal starter in natural kimchi fermentation, but limited research has been conducted on its phages. In this study, prophage distribution and characterization in kimchi-derived Leuconostoc strains were investigated, and phage induction was performed. Except for one strain, 16 Leuconostoc strains had at least one prophage region with questionable and incomplete regions, which comprised 0.5-6.0% of the bacterial genome. Based on major capsid protein analysis, ten intact prophages and an induced incomplete prophage of Leu. lactis CBA3626 belonged to the Siphoviridae family and were similar to Lc-Nu-like, sha1-like, phiMH1-like, and TPA_asm groups. Bacterial immunology genes, such as superinfection exclusion proteins and methylase, were found on several prophages. One prophage of Leu. lactis CBA3626 was induced using mitomycin C and was confirmed as belonging to the Siphoviridae family. Homology of the induced prophage with 21 reported prophages was not high (< 4%), and 47% identity was confirmed only with TPA_asm from Siphoviridae sp. isolate ct3pk4. Therefore, it is suggested that Leuconostoc from kimchi had diverse prophages with less than 6% genome proportion and some immunological genes. Interestingly, the induced prophage was very different from the reported prophages of other Leuconostoc species.

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10Bacterial Community of Galchi-Baechu Kimchi Based on Culture-Dependent and - Independent Investigation and Selection of Starter Candidates

저자 : Tao Kim , Sojeong Heo , Hong-eun Na , Gawon Lee , Jong-hoon Kim , Mi-sun Kwak , Moon-hee Sung , Do-won Jeong

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 32권 3호 발행 연도 : 2022 페이지 : pp. 341-347 (7 pages)

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In this study, the bacterial community of galchi-baechu kimchi was determined using culture-based and culture-independent techniques (next generation sequencing:NGS), and showed discrepancies between results. Weissella koreensis and Pediococcus inopinatus were the dominant species according to the NGS results, while Bacillus species and P. inopinatus were dominant in the culture-dependent analysis. To identify safe starter candidates, sixty-five Bacillus strains isolated from galchi-baechu kimchi using culture-dependent methods were evaluated for their antibiotic resistance, presence of toxin genes, and hemolytic activity. Strains were then assessed for salt tolerance and protease and lipase activity. As a result, four strains-B. safensis GN5_10, B. subtilis GN5_19, B. velezensis GN5_25, and B. velezensis GT8-were selected as safe starter candidates for use in fermented foods.

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1Recent Advances in Synthetic, Industrial and Biological Applications of Violacein and Its Heterologous Production

저자 : Aqsa Ahmed , Abdullah Ahmad , Renhan Li , Waleed Al-ansi , Momal Fatima , Bilal Sajid Mushtaq , Samra Basharat , Ye Li , Zhonghu Bai

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1465-1480 (16 pages)

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Violacein, a purple pigment first isolated from a gram-negative coccobacillus Chromobacterium violaceum, has gained extensive research interest in recent years due to its huge potential in the pharmaceutic area and industry. In this review, we summarize the latest research advances concerning this pigment, which include (1) fundamental studies of its biosynthetic pathway, (2) production of violacein by native producers, apart from C. violaceum, (3) metabolic engineering for improved production in heterologous hosts such as Escherichia coli, Citrobacter freundii, Corynebacterium glutamicum, and Yarrowia lipolytica, (4) biological/pharmaceutical and industrial properties, (5) and applications in synthetic biology. Due to the intrinsic properties of violacein and the intermediates during its biosynthesis, the prospective research has huge potential to move this pigment into real clinical and industrial applications.

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2Duplex dPCR System for Rapid Identification of Gram-Negative Pathogens in the Blood of Patients with Bloodstream Infection: A Culture-Independent Approach

저자 : Juyoun Shin , Sun Shin , Seung-hyun Jung , Chulmin Park , Sung-yeon Cho , Dong-gun Lee , Yeun-jun Chung

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1481-1489 (9 pages)

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Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.

KCI등재SCOUPUS

3Changes in the Microbiome of Vaginal Fluid after Menopause in Korean Women

저자 : Sukyung Kim , Hoonhee Seo , Md Abdur Rahim , Saebim Lee , Yun-sook Kim , Ho-yeon Song

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1490-1500 (11 pages)

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Various microorganisms reside in the human vagina; the vaginal microbiome is closely linked to both vaginal and general health, and for this reason, microbiome studies of the vagina are an area of research. In this study, we analyzed the vaginal microbiome of women before and after menopause to further increase our understanding of the vaginal microbiome and its contribution to general health. We did a 16s rRNA gene-based metagenomic analysis on the vaginal fluids of 11 premenopausal and 19 postmenopausal women in Korea. We confirmed that the taxonomic composition was significantly different between the two groups. In postmenopausal women, species richness was significantly decreased, but species diversity was significantly increased. In particular, among the taxonomic components corresponding to all taxon ranks of the vaginal microbiome, a reduction in Lactobacillus taxa after menopause contributed the most to the difference between the two groups. In addition, we confirmed through metabolic analysis that the lactic-acid concentration was also decreased in the vaginal fluid of women after menopause. Our findings on the correlation between menopause and the microbiome could help diagnose menopause and enhance the prevention and treatment diseases related to menopause.

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4Lagerstroemia ovalifolia Exerts Anti-Inflammatory Effects in Mice of LPS-Induced ALI via Downregulating of MAPK and NF-κB Activation

저자 : Jae-hong Min , Seong-man Kim , Ji-won Park , Nam Hoon Kwon , Soo Hyeon Goo , Ngatinem , Sri Ningsih , Jin-hyub Paik , Sangho Choi , Sei-ryang Oh , Sang-bae Han , Kyung-seop Ahn , Jae-won Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1501-1507 (7 pages)

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Lagerstroemia ovalifolia Teijsm. & Binn. (LO) (crape myrtle) has reportedly been used as traditional herbal medicine (THM) in Java, Indonesia. Our previous study revealed that the LO leaf extract (LOLE) exerted anti-inflammatory effects on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Based on this finding, the current study aimed to evaluate the protective effects of LOLE in a mouse model of LPS-induced acute lung injury (ALI). The results showed that treatment with LPS enhanced the inflammatory cell influx into the lungs and increased the number of macrophages and the secretion of the inflammatory cytokines in the bronchoalveolar lavage fluid (BALF) of mice. However, these effects were notably abrogated with LOLE pretreatment. Furthermore, the increase of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and monocyte chemoattractant protein-1 (MCP-1) expression in the lung tissues of mice with ALI was also reversed by LOLE. In addition, LOLE significantly suppressed the LPS-induced activation of the MAPK/NF-κB signaling pathway and led to heme oxygenase-1 (HO-1) induction in the lungs. Additionally, in vitro experiments showed that LOLE enhanced the expression of HO-1 in RAW264.7 macrophages. The aforementioned findings collectively indicate that LOLE exerts an ameliorative effect on inflammatory response in the airway of ALI mice.

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5Hsa_Circ_0001947/MiR-661/DOK7 Axis Restrains Non-Small Cell Lung Cancer Development

저자 : Yuyan Bao , Yanjie Yu , Bing Hong , Zhenjian Lin , Guoli Qi , Jie Zhou , Kaiping Liu , Xiaomin Zhang

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1508-1518 (11 pages)

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Hsa_circ_0001947 is associated with multiple cancers, but its function in non-small cell lung cancer (NSCLC) is ambiguous and needs further research. The targeting relationship among circ_0001947, miR-661, and downstream of tyrosine kinase 7 (DOK7) was predicted by database and further verified by dual-luciferase reporter assay, while their expressions in cancer tissues and cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). After transfection, cell biological behaviors and expressions of miRNAs, miR-661 and DOK7 were determined by cell function experiments and qRT-PCR, respectively. Circ_0001947 was low-expressed in NSCLC tissues and cells. Circ_0001947 knockdown intensified cell viability and proliferation, induced cell cycle arrest at S phase, suppressed apoptosis and evidently enhanced miR-510, miR-587, miR-661 and miR-942 levels, while circ_0001947 overexpression did the opposite. MiR-661 was a target gene of circ_0001947 that participated in the regulation of circ_0001947 on cell biological behaviors. Furthermore, DOK7, the target gene of miR-661, partly participated in the regulation of miR-661 on cell viability. Hsa_circ_0001947 acts as a sponge of miR-661 to repress NSCLC development by elevating the expression of DOK7.

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6Direct and Indirect Reduction of Cr(VI) by Fermentative Fe(III)-Reducing Cellulomonas sp. Strain Cellu-2a

저자 : Anamika Khanal , Hor-gil Hur , James K. Fredrickson , Ji-hoon Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1519-1525 (7 pages)

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Hexavalent chromium (Cr(VI)) is recognized to be carcinogenic and toxic and registered as a contaminant in many drinking water regulations. It occurs naturally and is also produced by industrial processes. The reduction of Cr(VI) to Cr(III) has been a central topic for chromium remediation since Cr(III) is less toxic and less mobile. In this study, fermentative Fe(III)-reducing bacterial strains (Cellu-2a, Cellu-5a, and Cellu-5b) were isolated from a groundwater sample and were phylogenetically related to species of Cellulomonas by 16S rRNA gene analysis. One selected strain, Cellu-2a showed its capacity of reduction of both soluble iron (ferric citrate) and solid iron (hydrous ferric oxide, HFO), as well as aqueous Cr(VI). The strain Cellu-2a was able to reduce 15 μM Cr(VI) directly with glucose or sucrose as a sole carbon source under the anaerobic condition and indirectly with one of the substrates and HFO in the same incubations. The heterogeneous reduction of Cr(VI) by the surface-associated reduced iron from HFO by Cellu-2a likely assisted the Cr(VI) reduction. Fermentative features such as large-scale cell growth may impose advantages on the application of bacterial Cr(VI) reduction over anaerobic respiratory reduction.

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7Endophytic Fungi of Salt-Tolerant Plants: Diversity and Ability to Promote Plant Growth

저자 : Irina Khalmuratova , Doo-ho Choi , Jong-guk Kim , In-seon Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1526-1532 (7 pages)

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Suaeda australis, Phragmites australis, Suaeda maritima, Suaeda glauca Bunge, and Limonium tetragonum in the Seocheon salt marsh on the west coast of the Korean Penincula were sampled in order to identify the endophytes inhabiting the roots. A total of 128 endophytic fungal isolates belonging to 31 different genera were identified using the fungal internal transcribed spacer (ITS) regions and the 5.8S ribosomal RNA gene. Fusarium, Paraconiothyrium and Alternaria were the most commonly isolated genera in the plant root samples. Various diversity indicators were used to assess the diversity of the isolated fungi. Pure cultures containing each of the 128 endophytic fungi, respectively, were tested for the plant growth-promoting abilities of the fungus on Waito-C rice germinals. The culture filtrate of the isolate Lt-1-3-3 significantly increased the growth of shoots compared to the shoots treated with the control. Lt-1-3-3 culture filtrate was analyzed and showed the presence of gibberellins (GA1 2.487 ng/ml, GA3 2.592 ng/ml, GA9 3.998, and GA24 6.191 ng/ml). The culture filtrate from the Lt-1-3-3 fungal isolate produced greater amounts of GA9 and GA24 than the wild-type Gibberella fujikuroi, a fungus known to produce large amounts of gibberellins. By the molecular analysis, fungal isolate Lt-1-3-3 was identified as Gibberella intermedia, with 100% similarity.

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8Genome-Wide Transcriptomic Analysis of n-Caproic Acid Production in Ruminococcaceae Bacterium CPB6 with Lactate Supplementation

저자 : Shaowen Lu , Hong Jin , Yi Wang , Yong Tao

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1533-1544 (12 pages)

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n-Caproic acid (CA) is gaining increased attention due to its high value as a chemical feedstock. Ruminococcaceae bacterium strain CPB6 is an anaerobic mesophilic bacterium that is highly prolific in its ability to perform chain elongation of lactate to CA. However, little is known about the genome-wide transcriptional analysis of strain CPB6 for CA production triggered by the supplementation of exogenous lactate. In this study, cultivation of strain CPB6 was carried out in the absence and presence of lactate. Transcriptional profiles were analyzed using RNA-seq, and differentially expressed genes (DEGs) between the lactate-supplemented cells and control cells without lactate were analyzed. The results showed that lactate supplementation led to earlier CA p,roduction, and higher final CA titer and productivity. 295 genes were substrate and/or growth dependent, and these genes cover crucial functional categories. Specifically, 5 genes responsible for the reverse β-oxidation pathway, 11 genes encoding ATP-binding cassette (ABC) transporters, 6 genes encoding substrate-binding protein (SBP), and 4 genes encoding phosphotransferase system (PTS) transporters were strikingly upregulated in response to the addition of lactate. These genes would be candidates for future studies aiming at understanding the regulatory mechanism of lactate conversion into CA, as well as for the improvement of CA production in strain CPB6. The findings presented herein reveal unique insights into the biomolecular effect of lactate on CA production at the transcriptional level.

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9Overexpression of Capsular Polysaccharide Biosynthesis Protein in Lactobacillus plantarum P1 to Enhance Capsular Polysaccharide Production for Di-n-butyl Phthalate Adsorption

저자 : Wei-bing Liu , Zhi-wei Lin , Ying Zhou , Bang-ce Ye

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1545-1551 (7 pages)

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Exopolysaccharides (EPSs) such as capsular polysaccharide (CPS) are important bioactive carbohydrate compounds and are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPS-producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs having many different structures and no health risks. However, EPS production by LAB does not meet the needs of large-scale application on an industrial scale. Here, the capA gene (encoding CPS biosynthesis protein) was overexpressed in Lactobacillus plantarum P1 to improve the production of EPSs and further enhance the DBP adsorption capability. Compared with P1, the CPS production in capA overexpressed strain was increased by 11.3 mg/l, and the EPS thickness was increased from 0.0786 ± 0.0224 μm in P1 to 0.1160 ± 0.0480 μm in P1-capA. These increases caused the DBP adsorption ratio of P1-capA to be doubled. Overall, the findings in this study provide a safe method for the adsorption and removal of DBP.

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10Effect of Enterotoxigenic Escherichia coli on Microbial Communities during Kimchi Fermentation

저자 : Woojung Lee , Hyo Ju Choi , Hyunwoo Zin , Eiseul Kim , Seung-Min Yang , Jinhee Hwang , Hyo-sun Kwak , Soon Han Kim , Hae-yeong Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 11호 발행 연도 : 2021 페이지 : pp. 1552-1558 (7 pages)

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The diverse microbial communities in kimchi are dependent on fermentation period and temperature. Here, we investigated the effect of enterotoxigenic Escherichia coli (ETEC) during the fermentation of kimchi at two temperatures using high-throughput sequencing. There were no differences in pH between the control group, samples not inoculated with ETEC, and the ETEC group, samples inoculated with ETEC MFDS 1009477. The pH of the two groups, which were fermented at 10 and 25℃, decreased rapidly at the beginning of fermentation and then reached pH 3.96 and pH 3.62. In both groups, the genera Lactobacillus, Leuconostoc, and Weissella were predominant. Our result suggests that microbial communities during kimchi fermentation may be affected by the fermentation parameters, such as temperature and period, and not enterotoxigenic E. coli (ETEC).

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