APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2024
·
IF 3.9
Xuemeng Ji,Dancai Fan,Jin Wang,Bowei Zhang,Yaozhong Hu,Huan Lv,Jing Wu,Yi Sun,Jingmin Liu,Yan Zhang,Shuo Wang
Cronobacter sakazakii poses a significant threat, particularly to neonates and infants. Despite its strong pathogenicity, understanding of C. sakazakii biofilms and their role in infections remains limited. This study investigates the roles of HmsP and c-di-GMP in biofilm formation and identifies key genetic and proteomic elements involved. Gene knockout experiments reveal that HmsP and c-di-GMP are linked to biofilm formation in C. sakazakii. Comparative proteomic profiling identifies the lysozyme inhibitor protein LprI, which is downregulated in hmsP knockouts and upregulated in c-di-GMP knockouts, as a potential biofilm formation factor. Further investigation of the lprI knockout strain shows significantly reduced biofilm formation and decreased virulence in a rat infection model. Additionally, LprI is demonstrated to bind extracellular DNA, suggesting a role in anchoring C. sakazakii within the biofilm matrix. These findings enhance our understanding of the molecular mechanisms underlying biofilm formation and virulence in C. sakazakii, offering potential targets for therapeutic intervention and food production settings.
研究领域:
微生物学
·
分子生物学
·
感染病学
✓ 已验证
DOI
PubMed
高影响因子
MICROBIOLOGICAL RESEARCH
·
2023
·
IF 6.7
Xiao-Yu Sun,Jie Deng,Chenhui Zhang,Sin-Yee Fung,Kam-Leung Siu,Ying-Ying Cheng,Liumei Ye,Jiaoxia Qin,Ke Wang,Jiu-Xin Qu,Wenying Gao,Fuxiang Wang,Dong-Yan Jin,Liang Yang
C-di-GMP is a bacterial second messenger implicated in the regulation of many key functions including antibiotic tolerance and biofilm formation. Our understanding of how c-di-GMP exerts its action via receptors to modulate different biological functions is still limited. Here we used a c-di-GMP affinity pull-down assay coupled to LC-MS/MS to identify c-di-GMP-binding proteins in the opportunistic pathogen Stenotrophomonas maltophilia. This analysis identified Smlt3238 (SodA), a protein of the superoxide dismutase family, as a c-di-GMP-binding protein. Microscale thermophoresis showed that purified SodA protein bound c-di-GMP with an estimated dissociation constant (Kd) value of 141.5 μM. Using various in vivo and in vitro experiments, we demonstrated that c-di-GMP modulates the enzyme activity of SodA directly. Circular dichroism experiments revealed that SodA protein gradually altered its basic structure with increasing levels of c-di-GMP. Phenotypic experiments conducted in the presence of a range of intracellular c-di-GMP levels showed that SodA function is modulated by c-di-GMP. The findings thus identify a novel c-di-GMP binding protein that governs oxidative stress tolerance in S. maltophilia .
研究领域:
微生物学
·
分子生物学
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
Biotechnology Journal
·
2024
·Vol. 19
·期 2
·2300542
·
IF 3.2
Yongshuai Yang,Siyu Guo,Can-Jian Hong,Zhao-Xun Liang,Chun Loong Ho
Bacterial second messenger c-di-GMP upregulation is associated with the transition from planktonic to sessile microbial lifestyle, inhibiting cellular motility, and virulence. However, in-depth elucidation of the cellular processes resulting from c-di-GMP upregulation has not been fully explored. Here, we report the role of upregulated cellular c-di-GMP in promoting planktonic cell growth of Escherichia coli K12 and Pseudomonas aeruginosa PAO1. We found a rapid expansion of cellular growth during initial cellular c-di-GMP upregulation, resulting in a larger planktonic bacterial population. The initial increase in c-di-GMP levels promotes bacterial swarming motility during the growth phase, which is subsequently inhibited by the continuous increase of c-di-GMP, and ultimately facilitates the formation of biofilms. We demonstrated that c-di-GMP upregulation triggers key bacterial genes linked to bacterial growth, swarming motility, and biofilm formation. These genes are mainly controlled by the master regulatory genes csgD and csrA . This study provides us a glimpse of the bacterial behavior of evading potential threats through adapting lifestyle changes via c-di-GMP regulation.
研究领域:
微生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 104
·期 5
·2205-2216
·
IF 3.53
Wang Feifei,Wang Yongzheng,Cen Congnan,Fu Linglin,Wang Yanbo
Cyclic diguanylate (c-di-GMP) is a second messenger involved in the regulation of various physiological processes in bacteria. However, its function in spoilage bacteria has not yet been addressed. Here, we studied the function of a tandem GGDEF-EAL domain protein, Sbal_3235, in the spoilage bacterium Shewanella baltica OS155. The deletion of sbal_3235 significantly reduced the c-di-GMP level, biofilm formation, and exopolysaccharide, trimethylamine (TMA), and putrescine production; sbal_3235 deletion also downregulated the expression of the torS and speF genes and affected membrane fatty acid composition. Site-directed mutagenesis in conserved GGDEF and EAL motifs abolished diguanylate cyclase (DGC) and phosphodiesterase (PDE) activity, respectively. These data indicate that Sbal_3235 is an essential contributor to the c-di-GMP pool with bifunctional DGC and PDE activity, which is involved in the biofilm formation and spoilage activity of S. baltica OS155. Our findings expand the biochemical role of c-di-GMP and uncover its link to spoilage activities, providing novel targets for food quality and safety controlling.
研究领域:
微生物学
·
食品科学
✓ 已验证
DOI
PubMed
高影响因子
FOOD MICROBIOLOGY
·
2024
·
IF 4.5
Mengyuan Qin,Shuo Han,Miaomiao Chen,Pengju Li,Yuqi Wang,Wenfang Niu,Chao Gao,Hailei Wang,Yi Li
Hafnia paralvei , a Gram-negative foodborne pathogen, is found ubiquitously in various aquatic animals and seafoods, which can form biofilm as a dominant virulence factor that contributes to its pathogenesis. However, the biofilm formation mechanism of H. paralvei and its effect on food spoilage has not been fully characterized. Here we show that biofilm formation, is regulated by c-di-GMP which mediated by bcsB , can increase the spoilage ability of H. paralvei . We found that GTP was added exogenously to enhance the synthesis of c-di-GMP, which further promoted biofilm formation. The gene dgcC , one of 11 genes encoding GGDEF domain-containing proteins in H. paralvei , was significantly upregulated with GTP as substrate. The upregulation of dgcC contributes to a significant increase of c-di-GMP and the formation of biofilm. In addition, the overexpression of dgcC induced upregulation of bcsB , a reported effector protein encoding gene, which was further demonstrated that overexpression of bcsB can encourage the synthesis of bacterial cellulose and biofilm formation. The effect of biofilm formation induced by c-di-GMP on spoilage of Yellow River carp ( Cyprinus carpio ) was evaluated by sensory evaluation , the total viable count , and the total volatile basic nitrogen, which showed that biofilm formation can significantly increase the spoilage ability of H. paralvei on C. carpio . Our findings provide the regulation of c-di-GMP on expression of bcsB , that can contribute to biofilm formation and spoilage ability of H. paralvei , which is favor to understanding the pathogenesis of Hafnia paralvei and its role in food spoilage.
研究领域:
微生物学
·
食品安全
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Yingkun Shi,Zijun Niu,Argenis Rodas-Gonzalez,Songlei Wang,Yihua Wang,Lingfan Yang
Guanosine 5′-monophosphate (GMP) is an ubiquitous compound in food and contributes significantly to food flavour due to its prominent umami properties. In this study, the development of MXene/g-C 3 N 4 /Au-COOH was synthesised by electrostatic self-assembly. The strong adsorption-catalysis ability of nanocomposites was utilised to attach the GMP to the surface of electrode. The surface morphology, functional groups and chemical bonds were analysed via several techniques, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), fourier transform infrared (FT-IR), and x-ray photoelectron spectroscopy (XPS). Under the optimised conditions, the GMP concentration detected by the sensor had a linear relationship in the range of 0.1–480 μM (S/N = 3) with a low detection limit of 0.0292 μM. Meanwhile, the MXene/g-C 3 N 4 /Au-COOH/GCE has good selectivity, reproducibility and stability. The applicability of the sensor in meat had been successfully verified by four actual samples (chicken broth, fish broth, pork and beef). This study presented a possible strategy to the field of evaluation of meat flavour.
研究领域:
食品科学
·
材料科学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2020
·
IF 4.389
Shanshan Yang,Yichao Wu,Chenchen Qu,Jeremy B. Fein,Yizhuang He,Qiaoyun Huang,Peng Cai
Cyclic diguanylate monophosphate (c-di-GMP) is a ubiquitous intracellular secondary messenger which governs the transition from a bacterial cell’s planktonic state to biofilm formation by stimulating the production of a variety of exopolysaccharide material by the bacterial cell. A range of genes involved in c-di-GMP signaling in the Gram-negative species Comamonas testosteroni have been identified previously, yet the physical-chemical properties of the produced extracellular polymeric substances (EPS) and the bacterial adhesion characteristics regulated by c-di-GMP are not well understood. Here, we modulated the in vivo c-di-GMP levels of Comamonas testosteroni WDL7 through diguanylate cyclase (YedQ) and phosphodiesterase (YhjH) gene editing. The strains and their adhesion properties were characterized by Fourier-transform infrared and two-dimensional correlation spectroscopy analysis (FTIR-2D CoS), contact angle and zeta potential measurements, atomic force microscopy (AFM) and extended-Derjaguin-Landau-Verwey-Overbeek (ExDLVO) analysis. Our results show that high c-di-GMP levels promoted the secretion of long-chain hydrophobic and electroneutral extracellular polysaccharides and proteins. The protein molecules on WDL7/pYedQ2 promoted the bacterial self-aggregation and adhesion onto negatively charged surfaces. In contrast, the reduction of intracellular c-di-GMP concentrations resulted in a nearly 80 % decrease in the adhesion of bacterial cells, although little change in the surface hydrophobicity or surface charge properties were observed for these cells relative to the wild type. These results indicate that the reduced adsorption of WDL7/YhjH that we observed may be caused by the flagellum-accelerated mobility at low c-di-GMP concentrations. Taken together, these results improve our mechanistic understanding of the effects of c-di-GMP in controlling bacterial physical-chemical properties and initial biofilm development.
研究领域:
微生物学
·
分子生物学
·
生物物理化学
✓ 已验证
DOI
高影响因子
LUMINESCENCE
·
2017
·Vol. 33
·期 1
·119-124
·
IF 1.509
Nan Liu,Juan Hao,Keying Cai,Mulan Zeng,Zhenzhong Huang,Lili Chen,Bingxian Peng,Ping Li,Li Wang,Yonghai Song
A novel ratiometric fluorescence nanosensor for superoxide anion (O 2 •− ) detection was designed with gold nanoparticles-bovine serum albumin (AuNPs-BSA)@terbium/guanosine monophosphate disodium (Tb/GMP) nanoscale coordination polymers (NCPs) (AuNPs-BSA@Tb/GMP NCPs). The abundant hydroxyl and amino groups of AuNPs-BSA acted as binding points for the self-assembly of Tb 3+ and GMP to form core-shell AuNPs-BSA@Tb/GMP NCP nanosensors. The obtained probe exhibited the characteristic fluorescence emission of both AuNPs-BSA and Tb/GMP NCPs. The AuNPs-BSA not only acted as a template to accelerate the growth of Tb/GMP NCPs, but also could be used as the reference fluorescence for the detection of O 2 •− . The resulting AuNPs-BSA@Tb/GMP NCP ratiometric fluorescence nanosensor for the detection of O 2 •− demonstrated high sensitivity and selectivity with a wide linear response range (14 nM–10 μM) and a low detection limit (4.7 nM).
研究领域:
分析化学
·
纳米技术
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2018
·
IF 3.226
Amjad Hussain Memon,Runsheng Ding,Qipeng Yuan,Hao Liang,Yun Wei
Enhancing activity of multiple enzymes by co-localization are key topics in bionanotechnology. Herein, we introduce a facile co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP) in the matrix of Cu (copper) and GMP (guanosine 5′-monophosphate). The multienzyme system by co-immobilizing GOx&HRP observed with high catalytic efficiency and stability compared with free GOx&HRP enzymes. GOx&HRP@Cu/GMP stability was observed 40% increased at pH 3 and thermal stability more than 70% determined at 90 ᵒ C. The storage stability was sustained 8 times higher than free enzymes after thirteen days and reusability after reacting eight times with fresh substrate retains 90% relative activity. Further, GOx&HRP@Cu/GMP demonstrated an excellent selectivity in the development of glucose biosensor and exhibited good resistance against the protease enzymes and organic solvents. The obtained results emphasize that co-immobilization of GOx&HRP in the matrix of Cu/GMP could be an efficient protocol for the biocatalyst encapsulation.
研究领域:
生物技术
·
酶工程
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
EJNMMI Radiopharmacy and Chemistry
·
2024
·Vol. 9
·期 1
·1-17
·
IF 4.4
Chen Lijuan,Li Xiaochen,Ge Yao,Li Huiqiang,Li Ruili,Song Xiaosheng,Liang Jianfei,Zhang Weifeng,Li Xiaona,Wang Xiaoqi,Wang Yunjuan,Wu Yaping,Bai Yan,Wang Meiyun
Background A novel positron emission tomography (PET) imaging tracer, [ 18 F] SynVesT-1, targeting synaptic vesicle glycoprotein 2 (SV2A), has been developed to meet clinical demand. Utilizing the Trasis AllinOne-36 (AIO) module, we’ve automated synthesis to Good Manufacturing Practice (GMP) standards, ensuring sterile, pyrogen-free production. The fully GMP-compliant robust synthesis of [ 18 F] SynVesT-1 boosting reliability and introducing a significant degree of simplicity and its comprehensive validation for routine human use. Results [ 18 F] SynVesT-1 was synthesized by small modifications to the original [ 18 F] SynVesT-1 synthesis protocol to better fit AIO module using an in-house designed cassette and sequence. With a relatively small precursor load of 5 mg, [ 18 F] SynVesT-1 was obtained with consistently high radiochemical yields (RCY) of 20.6 ± 1.2% (the decay-corrected RCY, n = 3) at end of synthesis. Each of the final formulated batches demonstrated radiochemical purity (RCP) and enantiomeric purity surpassing 99%. The entire synthesis process was completed within a timeframe of 80 min (75 ± 3.1 min, n = 3), saves 11 min compared to reported GMP automated synthesis procedures. The in-human PET imaging of total body PET/CT and time-of-flight (TOF) PET/MR showed that [ 18 F] SynVesT-1 is an excellent tracer for SV2A. It is advantageous for decentralized promotion and application in multi-center studies. Conclusion The use of AIO synthesizer maintains high production yields and increases reliability, reduces production time and allows rapid training of production staff. Besides, the as-prepared [ 18 F] SynVesT-1 displays excellent in vivo binding properties in humans and holds great potential for the imaging and quantification of synaptic density in vivo.
研究领域:
医学影像学
·
神经生物学
·
放射化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Cell Reports
·
2025
·
IF 6.9
Mingfang Wang,Jia Zeng,Binbin Cui,Guangliang Wu,Xiayu Chen,Yikai Fu,Bing Gu,Yinyue Deng
It was previously demonstrated that intestinal pathogens have evolved different mechanisms to enhance their infection and colonization in the intestines of hosts. However, it is unclear how Shigella effectively survives in the presence of bile salts. Here, we report that the biofilm formation and pathogenicity of S . sonnei are induced by primary bile salts through the two-component system EvgS/EvgA. The response regulator EvgA controls the transcription of the cyclic AMP synthase-encoding gene. Furthermore, the effector protein CRP of the cyclic AMP (cAMP) signal not only positively controls the transcription of ydeH , a gene encoding cyclic di-GMP (c-di-GMP) synthase, but also forms a complex with YdeH to improve its catalytic production of c-di-GMP. Additionally, interacting with YdeH enhances the ability of CRP to bind to the promoter of ydeH . Our work provides insights into how S . sonnei utilizes cascade amplification of c-di-GMP to promote fitness and pathogenicity in response to primary bile salts.
研究领域:
微生物学
·
分子生物学
·
感染与免疫
·
信号转导
·
病原生物学
·
环境应答
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2024
·
IF 3.3
Xinli Peng,Siqi Wang,Kaiyue Su,Yufeng Sun,Zhixiang Xu
BACKGROUND Histamine may lead to low blood pressure, skin flushing and edema when it accumulates in large amounts in the body. Therefore, establishing sensitive methods for the detection of histamine in foods is extremely important to ensure food safety and human health. RESULTS The MI-Cu-GMP NPs (2-methylimidazole-copper-guanosine monophosphate nanozymes) with high laccase-like activity were synthesized. Using the prepared molecular imprinted membrane as biomimetic antibody and MI-Cu-GMP NPs as marker, a sensitive direct competitive biomimetic enzyme-linked immunoassay (BELISA) method for rapid detection of the histamine in foods was developed. Under optimal conditions, the limit of detection (LOD, IC 15 ) and sensitivity (IC 50 ) of the BELISA method for histamine was 0.05 mg L −1 and 1.22 mg L −1 , respectively. The liquor samples spiked with histamine was detected by the BELISA method with satisfactory recoveries ranging from 90.00% to 116.00%. Further, the level of histamine in three samples (cooking wine, rice vinegar and soy sauce) was tested by the BELISA and high-performance liquid chromatography (HPLC), with no significant difference found between the two methods. CONCLUSION Given the advantages, the established BELISA method is expected to provide practical guidance for the monitoring of histamine in food and provides a foundation for the detection of other food hazards. © 2024 Society of Chemical Industry.
研究领域:
食品科学
·
生物化学
·
免疫分析
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）