APPLIED BIOCHEMISTRY AND MICROBIOLOGY
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2024
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IF 1
Hu J.,Wu Y.,Kang L.,Liu Y.,Wu Y.,Han D.,Wang J.
Abstract Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen that seriously threats the public and animal health. The major virulence of STEC is Shiga toxin (Stx) which is encoded by Stx prophage. Stx prophage induction leads to the production of Stx. Our recent research has shown that dietary functional sugars can differently regulate Shiga toxin type 2 (Stx2) prophage induction. However, high-throughput assays are required to find more dietary inhibitors for Stx prophage induction. This study aims to establish the recA dual-luciferase reporter system for rapidly screening the functional sugars that can inhibit Stx prophage induction. Results showed that the recA dual-luciferase reporter plasmid was successfully constructed as confirmed by DNA sequencing. Established recA dual-luciferase reporter system showed similar screening results when compared with traditional temperate phage determination method (i.e., qPCR) and suggested that L-arabinose is a dietary functional Stx prophage induction inhibitor. Supplementation with L-arabinose in culture medium reduces Stx production in STEC isolated from fecal samples of diarrhea piglets. Collectively, the recA dual-luciferase reporter system is a promising tool for convenient and rapid screening of dietary Stx prophage induction inhibitor, especially suitable for large-scale screening experiments.
研究领域:
微生物学
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分子生物学
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食品安全
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生物技术
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产品关联
DOI
高影响因子
ACS Applied Bio Materials
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2019
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IF 0
Huifan Li,Jiahui Zhong,Huijuan Zhu,Yuping Yang,Mengna Ding,Liulin Luo,Yuning Huo,Hexing Li
The hybrid Cu2O-TiO2 photocatalytic composite with a p-n heterojunction was synthesized by the supercritical solvothermal route. The uniformly distributed Cu2O was stably combined with TiO2 to benefit the increase of the specific surface area, the harvesting of visible light, and the separation of photogenerated holes and electrons. As a result, photocatalytic inactivation of Acinetobacter baumannii under visible-light irradiation was facilitated. Based on the photogenerated holes acting as the main active species, the Cu2O-TiO2 photocatalytic heterojunction could induce the leakage of K+ ions and serious damage of the cell structure, leading to the final cell death. During the photoantibacterial process on Cu2O-TiO2, the RecA gene played a significant role in enhancing the survival rate of Acinetobacter baumannii. Furthermore, the great bactericidal effect of Cu2O-TiO2 nanocomposites to different bacteria indicated the potential application for the environment-friendly disinfection.
研究领域:
微生物学
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环境科学
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材料科学
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DOI
PubMed
Environmental Science-Processes & Impacts
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2021
·Vol. 23
·期 7
·995-1005
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IF 4.238
Qing Ning,Dali Wang,Jing You
Quorum sensing inhibitors (QSIs) are promising alternatives to antibiotics. While QSIs have great application potential in a variety of fields, their joint effects with antibiotics on bacteria, especially on antibiotic resistance mutations, remain largely unexplored. Herein, we report the joint effects of four commonly used antibiotics and two QSIs on bacterial growth and resistance mutations in E. coli. It was found that QSIs presented antagonistic or additive effects with antibiotics on bacterial growth, and more importantly, QSIs exhibited an attenuating effect on antibiotic-induced resistance mutations. Further analysis demonstrated that antibiotics might enhance resistance mutations by promoting the expressions of rpoS, lexA and recA, while QSIs attenuated the mutations by promoting the expressions of mutS and uvrD. The present research provides a comprehensive understanding of the joint effects of antibiotics and QSIs on bacteria, which may benefit the risk assessment of their combined exposure.
研究领域:
微生物学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
npj Biofilms and Microbiomes
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2023
·Vol. 9
·期 1
·1-14
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IF 9.2
Hu Jie,Wu Yifan,Kang Luyuan,Liu Yisi,Ye Hao,Wang Ran,Zhao Jinbiao,Zhang Guolong,Li Xilong,Wang Junjun,Han Dandan
Elimination of specific enteropathogenic microorganisms is critical to gut health. However, the complexity of the gut community makes it challenging to target specific bacterial organisms. Accumulating evidence suggests that various foods can change the abundance of intestinal bacteria by modulating prophage induction. By using pathogenic Escherichia coli ( E. coli ) ATCC 25922 as a model in this research, we explored the potential of dietary modulation of prophage induction and subsequent bacterial survival. Among a panel of sugars tested in vitro, D -xylose was shown to efficiently induce prophages in E. coli ATCC 25922, which depends, in part, upon the production of D -lactic acid. In an enteric mouse model, prophage induction was found to be further enhanced in response to propionic acid. Dietary D -xylose increased the proportion of Clostridia which converted D -lactic acid to propionic acid. Intestinal propionic acid levels were diminished, following either oral gavage with the dehydrogenase gene ( ldhA )-deficient E. coli ATCC 25922 or depletion of intestinal Clostridia by administration of streptomycin. D -Xylose metabolism and exposure to propionic acid triggered E. coli ATCC 25922 SOS response that promoted prophage induction. E. coli ATCC 25922 mutant of RecA, a key component of SOS system, exhibited decreased phage production. These findings suggest the potential of using dietary components that can induce prophages as antimicrobial alternatives for disease control and prevention by targeted elimination of harmful gut bacteria.
研究领域:
微生物学
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胃肠病学
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营养学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
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2020
·Vol. 43
·期 20
·3840-3846
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IF 2.878
Changheng Jing,Fangzhi Yu,Ning Zhang,Yan Liu,Hailin Wang
Adenosine triphosphate is a universal energy currency that can directly provide energy required for a multitude of biochemical reactions and biophysical actions through adenosine triphosphatase catalyzed hydrolysis. Adenosine triphosphatase activity is thus one important feature for the characterization of protein function and cell activity. Herein, we optimized ion-pair reversed-phase high-performance liquid chromatography technique for highly efficient separation of adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate, and the method demonstrated good linearity. Moreover, by coupling a protein-removable ultrafiltration, we developed a sensitive and robust approach for quantification of adenosine triphosphatase hydrolytic activity. By this assay, we demonstrated that RecA filaments-catalyzed adenosine triphosphate hydrolysis approached a second-order reaction, and its rate constant was estimated as 0.057 mM −1 min −1 . In addition, we explored the effects of DNA length on this reaction and revealed that the increase of the length of single-stranded DNA can promote the adenosine triphosphatase hydrolytic activity of RecA filaments. All these results confirm the feasibility of this new method in quantification of adenosine triphosphatase hydrolytic activity assays. Compared with previous complicated enzyme-coupled or homogeneous colorimetric measurements, the developed approach with high resolution separation allows a simple reaction system for adenosine triphosphatase assay and a sensitive detection free of interference from background noise.
研究领域:
生物化学
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
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2019
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IF 5.714
Pan-Pan Jia,Tai Sun,Muhammad Junaid,Li Yang,Yan-Bo Ma,Zhi-Song Cui,Da-Peng Wei,Hao-Fei Shi,De-Sheng Pei
Graphene family nanomaterials (GFNs) have attracted significant attention due to their unique characteristics and applications in the fields of biomedicine and nanotechnology. However, previous studies highlighted the in vitro and in vivo toxicity of GFNs with size and oxidation state differences are still elusive. Therefore, we prepared graphene (G) and graphene oxide (GO) of three different sizes (S-small, M-medium, and L-large), and characterized them using multiple surface-sensitive analytical techniques. In vitro assays using HEK 293T cells revealed that the small and large sizes of G and GO significantly reduced the cell viability and increased DNA damage, accompanying with activated reactive oxygen species (ROS) generation and induced various expressions of associated critical genetic markers. Moreover, the bacterial assays highlighted that G and GO caused strong acute toxicity on Tox2 bacteria. Effects of G were higher than GO and showed size dependent effect: L > M > S, while the medium size of GO induced mild genetic toxicity on RecA bacteria. In vivo assays revealed that exposure to G and GO caused the developmental toxicity, induced ROS generation, and activated related pathways (specifically GO) in zebrafish. Taken together, G showed stronger ability to decrease the survival rate and induce the acute toxicity, while GO showed obvious toxicity in terms of DNA damages, ROS generation, and abnormal gene expressions. Our findings highlighted that G and GO differentially induced toxicity based on their varying physical characteristics, especially sizes and oxidation state, and exposure concentrations and sensitivity of the employed in vitro and in vivo models. In short, this study provided deep insights on the negative effects of GFNs exposure.
研究领域:
纳米技术
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生物医学
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毒理学
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细胞生物学
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环境科学
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产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2023
·
IF 8.1
Juan Xue,Jun Lv,Lanfang Liu,Fangfang Duan,Aiying Shi,Xuemeng Ji,Li Ding
Cronobacter sakazakii ( C. sakazakii ) is a notorious pathogen responsible for infections in infants and newborns, often transmitted through contaminated infant formula. Despite the use of traditional pasteurization methods, which can reduce microbial contamination, there remains a significant risk of pathogenic C. sakazakii surviving due to its exceptional stress tolerance. In our study, we employed a comparative proteomic approach by comparing wild-type strains with gene knockout strains to identify the essential genes crucial for the successful survival of C. sakazakii during desiccation. Our investigation revealed the significance of envZ-ompR , recA , and flhD gene cassettes in contributing to desiccation tolerance in C. sakazakii . Furthermore, through our comparative proteomic profiling, we identified the maltodextrin-binding protein encoded by ESA_03421 as a potential factor influencing dry tolerance. This protein is regulated by EnvZ-OmpR , RecA , and FlhD . Notably, the knockout of ESA_03421 resulted in a 150% greater reduction in Log CFU compared to the wild-type C. sakazakii . Overall, our findings offer valuable insights into the mechanisms underlying C. sakazakii desiccation tolerance and provide potential targets for the development of new antimicrobial strategies aimed at reducing the risk of infections in infants and newborns.
研究领域:
微生物学
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蛋白质组学
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食品安全
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婴儿健康
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产品关联
DOI
PubMed
高影响因子
Gut Microbes
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2023
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IF 12.2
Jie Hu,Yifan Wu,Xingjian Zhou,Luyuan Kang,Shiyi Zhang,Yisi Liu,Yu Pi,Xilong Li,Junjun Wang,Dandan Han
The pathogenicity of Escherichia coli (E. coli) O157:H7 is predominantly associated with Shiga toxin 2 (Stx2) that poses a huge threat to human and animal intestinal health. Production of Stx2 requires expression of stx2 gene, which is located in the genome of lambdoid Stx2 prophage. Growing evidence has implicated that many commonly consumed foods participate in the regulation of prophage induction. In this study, we aimed to explore whether specific dietary functional sugars could inhibit Stx2 prophage induction in E. coli O157:H7, thereby preventing Stx2 production and promoting intestinal health. We demonstrated that Stx2 prophage induction in E. coli O157:H7 was strongly inhibited by L-arabinose both in vitro and in a mouse model. Mechanistically, L-arabinose at doses of 9, 12, or 15 mM diminished RecA protein levels, a master mediator of the SOS response, contributing to reduced Stx2-converting phage induction. L-Arabinose inhibited quorum sensing and oxidative stress response, which are known as positive regulators of the SOS response and subsequent Stx2 phage production. Furthermore, L-arabinose impaired E. coli O157:H7 arginine transport and metabolism that were involved in producing Stx2 phage. Collectively, our results suggest that L-arabinose may be exploited as a novel Stx2 prophage induction inhibitor against E. coli O157:H7 infection.
研究领域:
微生物学
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食品科学
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分子生物学
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病理学
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产品关联
DOI
PubMed
高影响因子
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