ACS Synthetic Biology
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2021
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IF 5.11
Xinyi Cheng,Lu Pu,Shengwei Fu,Aiguo Xia,Shuqiang Huang,Lei Ni,Xiaochen Xing,Shuai Yang,Fan Jin
Bacterial pathogens operate by tightly controlling the pathogenicity to facilitate invasion and survival in host. While small molecule inducers can be designed to modulate pathogenicity to perform studies of pathogen–host interaction, these approaches, due to the diffusion property of chemicals, may have unintended, or pleiotropic effects that can impose limitations on their use. By contrast, light provides superior spatial and temporal resolution. Here, using optogenetics we reengineered GacS of the opportunistic pathogen Pseudomonas aeruginosa, signal transduction protein of the global regulatory Gac/Rsm cascade which is of central importance for the regulation of infection factors. The resultant protein (termed YGS24) displayed significant light-dependent activity of GacS kinases in Pseudomonas aeruginosa. When introduced in the Caenorhabditis elegans host systems, YGS24 stimulated the pathogenicity of the Pseudomonas aeruginosa strain PAO1 in a brain–heart infusion and of another strain, PA14, in slow killing media progressively upon blue-light exposure. This optogenetic system provides an accessible way to spatiotemporally control bacterial pathogenicity in defined hosts, even specific tissues, to develop new pathogenesis systems, which may in turn expedite development of innovative therapeutics.
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Scientific Reports
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2023
·Vol. 13
·期 1
·1-15
·
IF 4.6
Zheng Xiaoran,Gao Min,Wu Liangquan,Lu Xin,Lin Qiuqi,Zhong Hai,Lu Yingfei,Zhang Yunlei,Zhang Xiuwei
Pseudomonas aeruginosa ( P. aeruginosa ) infections present a grave threat to immunocompromised individuals, particularly those with cystic fibrosis due to the development of bacterial biofilms. In this study, we engineered self-assembling chitosan-ceftazidime nanoparticles (CSCE) capable of effectively penetrating biofilms and eradicating P. aeruginos a. The CSCE nanoparticles were synthesized through ionic cross-linking, combining negatively charged ceftazidime with positively charged chitosan, resulting in uniform nanoparticles measuring approximately 40 nm in diameter, exhibiting high dispersity and excellent biocompatibility. Remarkably, these nanoparticles exhibited significant inhibition of P. aeruginosa growth, reduced pyocyanin production, and diminished biofilm formation, achieving a maximum inhibition rate of 22.44%. Furthermore, in vivo investigations demonstrated enhanced survival in mice with abdominal P. aeruginosa infection following treatment with CSCE nanoparticles, accompanied by reduced levels of inflammatory cytokines Interleukin-6 (125.79 ± 18.63 pg/mL), Interleukin-17 (125.67 ± 5.94 pg/mL), and Tumor Necrosis Factor-α (135.4 ± 11.77 pg/mL). Critically, mice treated with CSCE nanoparticles showed no presence of bacteria in the bloodstream following intraperitoneal P. aeruginosa infection. Collectively, our findings highlight the potential of these synthesized nanoparticles as effective agents against P. aeruginosa infections.
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ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2019
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IF 4.527
Lin Li,Xianwei Shen,Chaocheng Zhao,Qiyou Liu,Xuwei Liu,Yanan Wu
A potent bacterial strain capable of degrading dibenzothiophene (DBT) was isolated and evaluated for its characteristics. The strain, designated as LKY-5, is rod-shaped, gram-negative, and occurs mainly in clusters. It was identified as belonging to the Pseudomonas genus based on the 16S rDNA sequence and phylogenic analysis. Determination of its DBT depletion efficiency by gas chromatography revealed that the isolate was able to completely degrade up to 100 mg L −1 DBT within 144 h. The pH values, DBT concentrations, and biomasses in the medium varied significantly in the initial 24 h. A biosurfactant produced by LKY-5 was extracted and identified as a di-rhamnolipid with the formula Rha-Rha-C 8 -C 8:1 by HPLC-ESI-MS/MS. There were 26 metabolites in the DBT degradation process. Pseudomonas sp. LKY-5 exhibited unusually high DBT degradation efficiency via multiple metabolic pathways. Compared with the reported 4S and Kodama pathways, two more expanded metabolic pathways for the degradation of DBT are proposed. The polycyclic aromatic sulfur heterocycles (PASHs) in diesel , such as C 1 -DBT, C 2 -DBT, C 3 -DBT, 4,6-DMDBT, and 2,4,6-TMDBT, can also be degraded with 28.2–42.3% efficiency. The results showed that LKY-5 is an excellent bacterial candidate for the bioremediation of PASH-contaminated sites and sediments.
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LWT-FOOD SCIENCE AND TECHNOLOGY
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2018
·
IF 3.129
Qisen Xiang,Chaodi Kang,Liyuan Niu,Dianbo Zhao,Ke Li,Yanhong Bai
In this work, the antibacterial effects and underlying mechanisms of plasma-activated water (PAW) against Pseudomonas deceptionensis CM2 isolated from spoiling chicken meat were investigated. The population of P. deceptionensis CM2 was significantly reduced approximately 5 log units within 10 min of exposure to PAW. The results of the scanning electron microscope clearly showed distinguishable morphostructural changes in P. deceptionensis CM2 as a consequence of PAW treatment. The permeability change in the outer and cytoplasmic membranes was evaluated by using 1- N -phenylnaphthylamine (NPN) and propidium iodide (PI), respectively. The fluorescence intensities of NPN and PI in P. deceptionensis CM2 cells were significantly increased after PAW treatment ( p < 0.05), indicating the disruption of the membrane permeability barrier. Following activation with plasma, PAW acquires an acidic pH and a higher oxidation reduction potential and contains aqueous reactive species, such as H 2 O 2 , nitrate, and nitrite anions, which play a crucial role in the inactivation process of P. deceptionensis CM2 cells. These findings indicate that PAW is a promising alternative to traditional sanitizers used in food and food-processing environments.
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ANALYTICAL CHEMISTRY
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2017
·
IF 6.32
Yong He,Mengyao Wang,Enci Fan,Hui Ouyang,Huan Yue,Xiaoxiao Su,Guojian Liao,Lin Wang,Shuguang Lu,Zhifeng Fu
A virulent bacteriophage highly specific to Pseudomonas aeruginosa (P. aeruginosa) was isolated from hospital sewage using a lambda bacteriophage isolation protocol. The bacteriophage, named as PAP1, was used to functionalize tosyl-activated magnetic beads to establish a bacteriophage-affinity strategy for separation and detection of viable P. aeruginosa. Recognition of the target bacteria by tail fibers and baseplate of the bacteriophage led to capture of P. aeruginosa onto the magnetic beads. After a replication cycle of about 100 min, the progenies lysed the target bacteria and released the intracellular adenosine triphosphate. Subsequently, firefly luciferase-adenosine triphosphate bioluminescence system was used to quantitate the amount of P. aeruginosa. This bacteriophage-affinity strategy for viable P. aeruginosa detection showed a linear range of 6.0 × 102 to 3.0 × 105 CFU mL–1, with a detection limit of 2.0 × 102 CFU mL–1. The whole process for separation and detection could be completed after bacteria capture, bacteriophage replication, and bacteria lysis within 2 h. Since the isolated bacteriophage recognized the target bacteria with very high specificity, the proposed strategy did not show any signal response to all of the tested interfering bacteria. Furthermore, it excluded the interference from inactivated P. aeruginosa because the bacteriophage could replicate only in viable cells. The proposed strategy had been applied for detection of P. aeruginosa in glucose injection, human urine, and rat plasma. In the further work, this facile bacteriophage-affinity strategy could be extended for detection of other pathogens by utilizing virulent bacteriophage specific to other targets.
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LWT-FOOD SCIENCE AND TECHNOLOGY
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2024
·
IF 6
Zitong Ge,Xiaxin Du,Jianan Liu,Junya Zhu,Hongshun Hao,Jingran Bi,Hongman Hou,Gongliang Zhang
Pseudomonas fluorescens , a specific spoilage organism, can control cell density and various physiological processes through quorum sensing (QS). Quorum sensing inhibitors (QSIs) are a crucial strategy for enhancing food quality and safety. In this study, benzyl isothiocyanate (BITC) at subminimum inhibitory concentrations (MICs) promoted protein and nucleic acid leakage, β-galactosidase activity and the relative fluorescence intensity of the membrane potential in P. fluorescens . N-butyryl- l -homoserine lactone (C4-HSL) and N-decanoyl- l -homoserine lactone (C10-HSL) were detected in P. fluorescens by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). C4-HSL and C10-HSL are known to orchestrate the QS of gram-negative bacteria. The inhibition rates of C4-HSL and C10-HSL by BITC at 1/2 MIC were 54.0%–96.2% and 31.6%–70.5%, respectively, after cultured with P. fluorescens for 6–24 h. The inhibitory effects of BITC at 1/4 MIC on P. fluorescens biofilm formation, extracellular polysaccharide production, and motility were enhanced in the presence of exogenous C4-HSL. Moreover, sub-MICs of BITC markedly downregulated the relative expression of QS-related genes , and the molecular docking results also showed excellent binding ability of BITC to QS proteins. This study demonstrated the feasibility of BITC as a QSI, whereby exogenous C4-HSL promoted the ability of BITC to disturb the QS system in P. fluorescens .
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ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2024
·
IF 6.2
Kejian Tian,Jinming Gu,Yibing Wang,Fenglin Zhang,Dandan Zhou,Qing Qiu,Yue Yu,Xuejian Sun,Menghan Chang,Xinwen Zhang,Hongliang Huo
Bisphenol A (BPA) is a globally concerning toxic pollutant, and microbial degradation is considered an effective method to treat BPA contamination. However, the inherent microbial toxicity of BPA is often overlooked, particularly the microbial mechanisms of resistance and detoxification against BPA. This study found that under the toxic stress of BPA, cbb3-type cytochrome c oxidase (cbb3-Cox) in the cells of Pseudomonas asiatica P1 ( P. asiatica P1) was the first to resist the toxicity. Genes such as ccoNOQPG showed significant upregulation with an average log 2 FC value of 3.56. Subsequently, genes that are related to metal ion binding, transport, and DNA repair were upregulated in the middle to later phase, which enhanced the metabolic functions of the strains and induced strain mutations to assist P. asiatica P1 in resisting the BPA toxicity. Meanwhile, three potential BPA degradation genes were identified, among which sdr P1 was crucial to the BPA degradation and detoxification. After genetic recombination, sdr P1 achieved a degradation rate of 92.52 % for BPA. Furthermore, through various methods such as alkyl interactions, sdr P1 exhibited oxidation and demethylation to form lower toxic intermediate products and complete the biological detoxification of BPA. This study provides a systematic analysis of the toxicity resistance, biodegradation, and detoxification processes in bacterial BPA removal, refines the mechanism of BPA biodegradation and contributes to a more comprehensive and systematic understanding of the overall process of microbial removal of toxic pollutants.
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STAR Protocols
·
2024
·
IF 1.3
Jiahui Huang,Yao Zhang,Shuo Sheng,Lian-Hui Zhang,Zeling Xu
Summary Pseudomonas aeruginosa utilizes a type III secretion system (T3SS) to directly inject effector proteins into host cells, leading to severe acute infections. Here, we present a protocol for detecting the T3SS-mediated cytotoxicity of P. aeruginosa using the A549 cell line. We describe the steps for the preparation of the A549 cell line and P. aeruginosa strains, cell seeding, bacterial culture, infection, and cytotoxicity assay. Additionally, we provide detailed procedures for data analysis. For complete details on the use and execution of this protocol, please refer to Huang et al. 1
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2021
·
IF 5.279
Hua Yang,Leqiong Jin,Dengqi Zhao,Zhifeng Lian,Manikandan Appu,Jianying Huang,Zibin Zhang
An omnipresent pathogenic bacterium, Pseudomonas aeruginosa (P. aeruginosa PAO1), is easy to contaminate environmental water or foods, causing daily food spoilage and infections. The biofilm-forming ability and bacterial resistance of P. aeruginosa PAO1 make it difficult to be eradicated by traditional bacteriostatic agents. In this work, we designed and synthesized a pyridinium-based pillar[5]arene (PP5), while trimethylammonium-based pillar[5]arene (TP5) was used as a control compound. After clear characterization, the antibacterial and antibiofilm activities as well as the microbial resistance of TP5 and PP5 against P. aeruginosa PAO1 were extensively examined. It was revealed that PP5 exhibited good inhibition activity with the minimum inhibitory concentration (MIC) of 0.051 mmol/L, while no significant antibacterial and biofilm formation activity for TP5 against P. aeruginosa PAO1 was observed. More importantly, PP5 had negligible antimicrobial resistance even after 18th passages. A transmission electron microscope (TEM) showed that PP5 could physically disrupt the cell membranes, causing the leakage of internal constituents, which is possibly ascribed to the synergistic penetrability and π–π interactions of strain, thus greatly reduced the development of bacterial resistance. Overall, the presented studies indicated that pyridinium moieties could facilitate the cationic pillar[5]arene to generate surprising antibacterial and antibiofilm formation ability against P. aeruginosa PAO1.
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Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Ma Anzhou,Ma Jianpeng,Chen Xianke,Zhuang Guoqiang
Highly chlorinated benzenes usually have a low efficient degradation in environment. Here we proposed a synergistic removal strategy of pentachlorobenzene (PeCB) using Pseudomonas sp. JS100 coupled with immobilized nanoscale zero-valent iron (NZVI). The structural and textural features of the synergistic system were characterized by X-ray powder diffraction, field emission scanning electron microscopy, and a specific surface area and pore size analysis. Nanoscale zero-valent iron particles were dispersed and attached to the biofilter, which increased the specific surface area to 34.5 m2 g−1. The batch experiment revealed that the removal efficiency of PeCB reached 80.2% in the synergistic system within 48 h. The degradation followed pseudo-first-order reaction kinetics, and the reaction rate constant was measured to be 0.0336 h−1. In the degradation mechanism, PeCB was degraded by NZVI to lower chlorobenzenes, which were utilized by Pseudomonas sp. JS100 as nutrients, thereby achieving rapid removal of PeCB.
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2014
·Vol. 99
·期 7
·3211-3220
·
IF 3.811
Wang Xiangqian,Wu Chao,Liu Nan,Li Sujing,Li Wei,Chen Jianmeng,Chen Dongzhi
A Pseudomonas sp. strain WL2 that is able to efficiently metabolize ethyl mercaptan (EM) into diethyl disulfide (DEDS) through enzymatic oxidation was isolated from the activated sludge of a pharmaceutical wastewater plant. One hundred percent removal of 113.5 mg L −1 EM and 110.3 mg L −1 DEDS were obtained within 14 and 32 h, respectively. A putative EM degradation pathway that involved the catabolism via DEDS was proposed, which indicated DEDS were further mineralized into carbon dioxide (CO 2 ), bacterial cells, and sulfate (SO 4
2− ) through the transformation of element sulfur and ethyl aldehyde. Degradation kinetics for EM and DEDS with different initial concentrations by strain WL2 were evaluated using Haldane-Andrews model with maximum specific degradation rates of 3.13 and 1.33 g g −1 h −1 , respectively, and maximum degradation rate constants of 0.522 and 0.175 h −1 using pseudo-first-order kinetic model were obtained. Results obtained that aerobic degradation of EM by strain WL2 was more efficient than those from previous studies. Substrate range studies of strain WL2 demonstrated its ability to degrade several mercaptans, disulfides, aldehydes, and methanol. All the results obtained highlight the potential of strain WL2 for the use in the biodegradation of volatile organic sulfur compounds (VOSCs).
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2020
·
IF 4.192
Miaomiao Li,Jinlong Song,Qingyun Ma,Delong Kong,Yiqing Zhou,Xu Jiang,Rebecca Parales,Zhiyong Ruan,Qinghua Zhang
A total of five strains of nicosulfuron-degrading bacteria were isolated from a continuously cultivated microbial consortium using culturomics. Among them, a novel Pseudomonas strain, LAM1902, with the highest degradation efficiency was investigated in detail. The characteristics of nicosulfuron-degradation by LAM1902 were investigated and optimized by response surface analysis. Furthermore, non-targeted metabolomic analysis of extracellular and intracellular biodegradation of nicosulfuron by LAM1902 was carried out by liquid chromatography/mass spectroscopy (LC–MS) and gas chromatography–time-of-flight/mass spectroscopy (GC–TOF/MS). It was found that nicosulfuron was degraded by LAM1902 mainly via breaking the sulfonylurea bridge, and this degradation might be attributed to oxalate accumulation. The results of GC–TOF/MS also showed that the intracellular degradation of nicosulfuron did not occur. However, nicosulfuron exerted a significant influence on the metabolism of inositol phosphate, pyrimidine, arginine/proline, glyoxylate, and dicarboxylate metabolism and streptomycin biosynthesis. The changes of myo-inositol, trehalose, and 3-aminoisobutanoic acid were proposed as a mechanism of self-protection against nicosulfuron stress.
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