PLoS One
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2019
·Vol. 14
·期 4
·e0215084
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IF 2.776
Xiaoqin Wang,Zhi-Min Li,Qingyue Li,Mingsong Shi,Lingling Bao,Dingguo Xu,Zhimin Li
Fermentation-respiration switch protein (FrsA) was thought to play an important role in controlling the metabolic flux between respiration and fermentation pathways, whereas the biochemical function of FrsA was unclear yet. A gene coding for FrsA protein from Vibrio vulnificus was chemically synthesized. The recombinant VvFrsA was expressed as a soluble protein and purified by Ni-NTA affinity chromatography. The protein had a subunit molecular weight of ca . 45 kDa by SDS-PAGE and preferred short-chain esters when p-nitrophenyl alkanoate esters were used as substrates. Optimum condition for VvFrsA was found to be at pH 9.0 and 50 °C. The protein retained high esterase activity at alkaline condition and would denature slowly at over 50 °C. With p-nitrophenyl acetate as the substrate, the K m and k cat were determined to be 18.6 mM and 0.67 s -1 , respectively, by steady-state kinetic assay. Molecular dynamics simulation and docking model structure revealed that p-nitrophenyl acetate could be the substrate of VvFrsA. In conclusion our results demonstrated that the protein was able to catalyze the hydrolysis of esters, especially p-nitrophenyl acetate, for the first time.
研究领域:
微生物学
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生物化学
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DOI
PubMed
高影响因子
Fishes
·
2024
·Vol. 9
·期 4
·131
·
IF 2.1
Yunji Xiu,Jingyuan Yi,Ruixin Feng,Jiaxue Song,Yunfei Pang,Peng Liu,Shun Zhou
TheAeromonas salmonicidais responsible for causing furunculosis in various fish species. Furunculosis is a ubiquitous disease that affects the aquaculture industry and causes the mass mortality of turbot.Vibrio vulnificusis a pathogen that causes skin ulcers and hemorrhagic septicemia in fish, resulting in significant mortality in aquaculture. In this study, we have established a bivalent inactivated vaccine againstA. salmonicidaandV. vulnificuswith Montanide™ ISA 763 AVG as an adjuvant. This bivalent inactivated vaccine was used to immunize turbot by intraperitoneal injection, and the relevant immune indexes were detected. The results demonstrate that the bivalent inactivated vaccine exhibited a relative percent survival (RPS) of 77% followingA. salmonicidaandV. vulnificusintraperitoneal challenge. The vaccinated group exhibited higher levels of acid phosphatase activity and lysozyme activity compared to the control group. ELISA results showed a significant increase in serum antibody levels in immunized turbot, which was positively correlated with immunity. In the kidney tissue, related immune genes (TLR5,CD4,MHCIandMHCII) were up-regulated significantly, showing that the vaccine can induce cellular and humoral immune responses in turbot. In conclusion, the bivalent inactivated vaccine againstA. salmonicidaandV. vulnificuswas immunogenic, efficiently preventing turbot from infection, which has the potential to be applied in aquaculture.
研究领域:
水产养殖
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免疫学
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鱼类疾病
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疫苗学
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DOI
高影响因子
Frontiers in Microbiology
·
2021
·
IF 5.64
Liu Xinning,Liu Yuanchang,Zhao Guangjian,Zhang Yidan,Liu Lu,Wang Juan,Wang Yifan,Zhang Siyu,Li Xin,Guo Dongliang,Wang Peng,Xu Ximing
Vibrio vulnificus is a zoonotic bacterium that is capable of causing highly lethal diseases in humans; this pathogen is responsible for 95% of all seafood-related deaths in the United States. Arylamine N-acetyltransferases (NAT, E.C. 2.3.1.5) is a major family of xenobiotic-metabolizing enzymes that can biotransform aromatic amine chemicals. In this research, to evaluate the effect of NAT on acetyl group transformation in arylamine antibiotics, we first used sequence alignment to study the structure of V. vulnificus NAT [(VIBVN)NAT]. The nat gene encodes a protein of 260 amino acids, which has an approximate molecular mass of 30 kDa. Then we purified recombinant (VIBVN)NAT and determined the enzyme activity by PNPA and DTNB methods. The DTNB method indicates that this prokaryotic NAT has a particular substrate specificity towards aromatic substrates. However, (VIBVN)NAT lost most of its activity after treatment with high concentrations of urea and H2O2. In addition, we also explored the stability of the enzyme at different temperatures and pH values. In analyzing the influence of metal ions, the enzyme activity was significantly inhibited by Zn2+ and Cu2+. The kinetic parameters Km and Vmax were determined using hydralazine, isoniazid, 4-amino salicylic acid, and 4-chloro-3-methylaniline as substrates, and the Tm, Tagg and size distribution of (VIBVN)NAT were observed. In particular, a molecular docking study on the structure of (VIBVN)NAT was conducted to understand its biochemical traits. These results showed that (VIBVN)NAT could acetylate various aromatic amine substrates and contribute to arylamine antibiotic resistance in V. vulnificus.
研究领域:
微生物学
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生物化学
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分子生物学
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药理学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Chenjing Xu,Jinpo Xie,Lian Yu,Bin Shu,Xiaogang Liu,Siping Chen,Qinglan Li,Shaohai Qi,Suqing Zhao
Vibrio vulnificus infection caused by contaminated aquatic products and seawater can lead to severe disease and high mortality. The development of a rapid and sensitive detection method for Vibrio vulnificus is vital to effectively prevent infection in advance. In this study, CeO 2 @PtRu with high peroxidase activity was used to construct a colorimetric immunoassay for Vibrio vulnificus detection by conjugating polyclonal antibodies via the biotin-streptavidin system. The developed colorimetric biosensor for Vibrio vulnificus demonstrated rapid operability and good sensitivity with a detection range from 10 4 CFU/mL to 10 9 CFU/mL, and the limit of detection (LOD) is 193 CFU/mL. Moreover, the colorimetric biosensor showed excellent specificity and good recoveries from 98.70% to 102.10% with RSD < 7.45% for spiked real samples. This novel CeO 2 @PtRu-based colorimetric biosensor has great application potential for the sensitive detection of Vibrio vulnificus in seafood.
研究领域:
微生物学
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免疫学
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生物传感技术
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DOI
PubMed
高影响因子
Polymers
·
2026
·Vol. 18
·期 6
·
IF 5.8
Fuyu Zhao,Xianjun Fu,Wuyi Zhou,Xia Ren
Wounds caused byVibrio vulnificus(V. vulnificus) infection often exhibit delayed healing and are prone to complications, making them a significant challenge in clinical treatment. Current conventional treatments, such as antibiotics and gauze dressings, have limited effectiveness. To address this, this study developed a multifunctional fiber membrane using electrospinning technology. Micron- or nano-sizedHaliotidis Concha(HC) and eugenol (Eu) were loaded onto the membrane to promote healing inV. vulnificus-infected wounds. The prepared fiber membranes exhibited diameters of approximately 0.35 ± 0.01 μm. Membranes loaded with nano-HC demonstrated significant antibacterial efficacy, achieving a 96.2% inhibition rate againstV. vulnificus, which was markedly superior to the micron-HC group (p< 0.05). Notably, the nano-HC/Eu membranes exhibited exceptionally high flexibility with an elongation at break of 878.1 ± 35.3%, while maintaining a tensile strength of approximately 2.2 MPa. Furthermore, these membranes exhibited excellent biocompatibility, with cell viability exceeding 85% for fibroblasts, and demonstrated good hemocompatibility. They also effectively promoted cell migration, indicating their potential as wound scaffold materials. In aV. vulnificus-infected skin wound model, the nano-HC/Eu fiber membrane accelerated collagen deposition and promoted wound healing, achieving a wound closure rate of 94.7 ± 1.1% on day 15. In summary, this study developed a multifunctional fiber membrane with antibacterial, antioxidant, and wound healing properties, offering a novel dressing for treatingV. vulnificusinfections.
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PubMed
高影响因子
FOOD CONTROL
·
2025
·
IF 6.3
Siping Chen,Li Ji,Jinpo Xie,Chenjing Xu,Qinglan Li,Suqing Zhao
Vibrio vulnificus have seriously threatened human health through wound infection and bacterial contamination of seafood. Herein, a novel fluorescent lateral flow immunoassay (LFIA) was developed to sensitively and rapidly detect Vibrio vulnificus by using polyclonal antibodies-functionalized CdTe@SiO 2 nanocomposite (CdTe@SiO 2 -pAb) as the advanced red fluorescent signal label. The fluorescent LFIA based on CdTe@SiO 2 -pAb exhibited a wide detection range from 2.51 × 10 4 CFU/mL to 1.40 × 10 8 CFU/mL with the low limit of detection of 3.95 × 10 3 CFU/mL for Vibrio vulnificus . Moreover, the constructed LFIA displayed excellent specificity and good recovery ranging from 93.3% to 104.1% towards spiked samples. Therefore, CdTe@SiO 2 -pAb was a promising fluorescent signal label of LFIA for the sensitive and handy detection of Vibrio vulnificus in contaminated seafood.
研究领域:
生物技术
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食品安全检测
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微生物学
✓ 已验证
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DOI
高影响因子
危险品化学品经营许可证（带存储）