scholarly journals Label-free, rapid and quantitative phenotyping of stress response in E. coli via ramanome

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Lin Teng ◽  
Xian Wang ◽  
Xiaojun Wang ◽  
Honglei Gou ◽  
Lihui Ren ◽  
...  
Keyword(s):  
Stress Response ◽  
Label Free ◽  
E Coli

scholarly journals Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherichia coli biofilm formation in co-culture

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Huiyi Song ◽  
Ni Lou ◽  
Jianjun Liu ◽  
Hong Xiang ◽  
Dong Shang
Keyword(s):  
Escherichia Coli ◽  
Proteomic Analysis ◽  
Biofilm Formation ◽  
Lactobacillus Rhamnosus ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Label Free ◽  
Quantitative Proteomic Analysis ◽  
E Coli ◽  
Protein Ubiquitination

Abstract Background Escherichia coli (E. coli) is the principal pathogen that causes biofilm formation. Biofilms are associated with infectious diseases and antibiotic resistance. This study employed proteomic analysis to identify differentially expressed proteins after coculture of E. coli with Lactobacillus rhamnosus GG (LGG) microcapsules. Methods To explore the relevant protein abundance changes after E. coli and LGG coculture, label-free quantitative proteomic analysis and qRT-PCR were applied to E. coli and LGG microcapsule groups before and after coculture, respectively. Results The proteomic analysis characterised a total of 1655 proteins in E. coli K12MG1655 and 1431 proteins in the LGG. After coculture treatment, there were 262 differentially expressed proteins in E. coli and 291 in LGG. Gene ontology analysis showed that the differentially expressed proteins were mainly related to cellular metabolism, the stress response, transcription and the cell membrane. A protein interaction network and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis indicated that the differentiated proteins were mainly involved in the protein ubiquitination pathway and mitochondrial dysfunction. Conclusions These findings indicated that LGG microcapsules may inhibit E. coli biofilm formation by disrupting metabolic processes, particularly in relation to energy metabolism and stimulus responses, both of which are critical for the growth of LGG. Together, these findings increase our understanding of the interactions between bacteria under coculture conditions.

Thermal and Starvation Stress Response of Escherichia coli O157:H7 Isolates Selected from Agricultural Environments

2016 ◽  
Vol 79 (10) ◽  
pp. 1673-1679 ◽  
Author(s):  
ACHYUT ADHIKARI ◽  
ANDY BARY ◽  
CRAIG COGGER ◽  
CALEB JAMES ◽  
GÜLHAN ÜNLÜ ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heat Stress ◽  
Stress Response ◽  
Weibull Model ◽  
Stress Conditions ◽  
Inactivation Kinetics ◽  
Escherichia Coli O157 ◽  
Starvation Stress ◽  
E Coli ◽  
Response To Stress

ABSTRACT Pathogens exposed to agricultural production environments are subject to multiple stresses that may alter their survival under subsequent stress conditions. The objective of this study was to examine heat and starvation stress response of Escherichia coli O157:H7 strains isolated from agricultural matrices. Seven E. coli O157:H7 isolates from different agricultural matrices—soil, compost, irrigation water, and sheep manure—were selected, and two ATCC strains were used as controls. The E. coli O157:H7 isolates were exposed to heat stress (56°C in 0.1% peptone water for up to 1 h) and starvation (in phosphate-buffered saline at 37°C for 15 days), and their survival was examined. GInaFiT freeware tool was used to perform regression analyses of the surviving populations. The Weibull model was identified as the most appropriate model for response of the isolates to heat stress, whereas the biphasic survival curves during starvation were fitted using the double Weibull model, indicating the adaptation to starvation or a resistant subpopulation. The inactivation time during heating to achieve the first decimal reduction time (δ) calculated with the Weibull parameters was the highest (45 min) for a compost isolate (Comp60A) and the lowest (28 min) for ATCC strain 43895. Two of the nine isolates (ATCC 43895 and a manure isolate) had β < 1, indicating that surviving populations adapted to heat stress, and six strains demonstrated downward concavity (β > 1), indicating decreasing heat resistance over time. The ATCC strains displayed the longest δ2 (>1,250 h) in response to starvation stress, compared with from 328 to 812 h for the environmental strains. The considerable variation in inactivation kinetics of E. coli O157:H7 highlights the importance of evaluating response to stress conditions among individual strains of a specific pathogen. Environmental isolates did not exhibit more robust response to stress conditions in this study compared with ATCC strains.

scholarly journals The Small Noncoding DsrA RNA Is an Acid Resistance Regulator in Escherichia coli

2004 ◽  
Vol 186 (18) ◽  
pp. 6179-6185 ◽  
Author(s):  
Richard A. Lease ◽  
Dorie Smith ◽  
Kathleen McDonough ◽  
Marlene Belfort
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Resistance Genes ◽  
Acid Resistance ◽  
Dna Arrays ◽  
E Coli ◽  
Specific Mrnas ◽  
Steady State Levels ◽  
Control Translation ◽  
K 12

ABSTRACT DsrA RNA is a small (87-nucleotide) regulatory RNA of Escherichia coli that acts by RNA-RNA interactions to control translation and turnover of specific mRNAs. Two targets of DsrA regulation are RpoS, the stationary-phase and stress response sigma factor (σs), and H-NS, a histone-like nucleoid protein and global transcription repressor. Genes regulated globally by RpoS and H-NS include stress response proteins and virulence factors for pathogenic E. coli. Here, by using transcription profiling via DNA arrays, we have identified genes induced by DsrA. Steady-state levels of mRNAs from many genes increased with DsrA overproduction, including multiple acid resistance genes of E. coli. Quantitative primer extension analysis verified the induction of individual acid resistance genes in the hdeAB, gadAX, and gadBC operons. E. coli K-12 strains, as well as pathogenic E. coli O157:H7, exhibited compromised acid resistance in dsrA mutants. Conversely, overproduction of DsrA from a plasmid rendered the acid-sensitive dsrA mutant extremely acid resistant. Thus, DsrA RNA plays a regulatory role in acid resistance. Whether DsrA targets acid resistance genes directly by base pairing or indirectly via perturbation of RpoS and/or H-NS is not known, but in either event, our results suggest that DsrA RNA may enhance the virulence of pathogenic E. coli.

scholarly journals Investigation of the Genes Involved in the Outbreaks of Escherichia coli and Salmonella spp. in the United States

Antibiotics ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1274
Author(s):  
Michelle Li ◽  
Kyle Wang ◽  
Ashley Tang ◽  
Aaron Tang ◽  
Andrew Chen ◽  
...  
Keyword(s):  
Stress Response ◽  
Antimicrobial Resistance ◽  
Pathogen Detection ◽  
Dimensional Space ◽  
Virulence Gene ◽  
Principal Component ◽  
The United States ◽  
Salmonella Spp ◽  
E Coli ◽  
Antimicrobial Resistance Genes

Salmonella spp. and Escherichiacoli (E. coli) are two of the deadliest foodborne pathogens in the US. Genes involved in antimicrobial resistance, virulence, and stress response, enable these pathogens to increase their pathogenicity. This study aims to examine the genes detected in both outbreak and non-outbreak Salmonella spp. and E. coli by analyzing the data from the National Centre for Biotechnology Information (NCBI) Pathogen Detection Isolates Browser database. A multivariate statistical analysis was conducted on the genes detected in isolates of outbreak Salmonella spp., non-outbreak Salmonella spp., outbreak E. coli, and non-outbreak E. coli. The genes from the data were projected onto a two-dimensional space through principal component analysis. Hierarchical clustering was then used to quantify the relationship between the genes in the dataset. Most of the outlier genes identified in E. coli isolates are virulence genes, while outlier genes identified in Salmonella spp. are mainly involved in stress response. Gene epeA, which encodes a high-molecular-weight serine protease autotransporter of Enterobacteriaceae (SPATE) protein, along with subA and subB that encode cytotoxic activity, may contribute to the pathogenesis of outbreak E. coli. The iro operon and ars operon may play a role in the ecological success of the epidemic clones of Salmonella spp. Concurrent relationships between esp and ter operons in E. coli and pco and sil operons in Salmonella spp. are found. Stress-response genes (asr, golT, golS), virulence gene (sinH), and antimicrobial resistance genes (mdsA and mdsB) in Salmonella spp. also show a concurrent relationship. All these findings provide helpful information for experiment design to combat outbreaks of E. coli and Salmonella spp.

scholarly journals Genetic analysis of the role of the conserved inner membrane protein CvpA in EHEC resistance to deoxycholate

2020 ◽  
Author(s):  
Alyson R. Warr ◽  
Rachel T. Giorgio ◽  
Matthew K. Waldor
Keyword(s):  
Stress Response ◽  
Membrane Protein ◽  
Bile Salt ◽  
Cell Envelope ◽  
Enteric Bacteria ◽  
Enteric Pathogens ◽  
Inner Membrane ◽  
Bacterial Phyla ◽  
E Coli ◽  
Inner Membrane Protein

The function of cvpA, a bacterial gene predicted to encode an inner membrane protein, is largely unknown. Early studies in E. coli linked cvpA to Colicin V secretion and recent work revealed that it is required for robust intestinal colonization by diverse enteric pathogens. In enterohemorrhagic E. coli (EHEC), cvpA is required for resistance to the bile salt deoxycholate (DOC). Here, we carried out genome-scale transposon-insertion mutagenesis and spontaneous suppressor analysis to uncover cvpA’s genetic interactions and identify common pathways that rescue the sensitivity of a ΔcvpA EHEC mutant to DOC. These screens demonstrated that mutations predicted to activate the σE-mediated extracytoplasmic stress response bypass the ΔcvpA mutant’s susceptibility to DOC. Consistent with this idea, we found that deletions in rseA and msbB and direct overexpression of rpoE restored DOC resistance to the ΔcvpA mutant. Analysis of the distribution of CvpA homologs revealed that this inner membrane protein is conserved across diverse bacterial phyla, in both enteric and non-enteric bacteria that are not exposed to bile. Together, our findings suggest that CvpA plays a role in cell envelope homeostasis in response to DOC and similar stress stimuli in diverse bacterial species. IMPORTANCE Several enteric pathogens, including Enterohemorrhagic E. coli (EHEC), require CvpA to robustly colonize the intestine. This inner membrane protein is also important for secretion of a colicin and EHEC resistance to the bile salt deoxycholate (DOC), but its function is unknown. Genetic analyses carried out here showed that activation of the σE-mediated extracytoplasmic stress response restored the resistance of a cvpA mutant to DOC, suggesting that CvpA plays a role in cell envelope homeostasis. The conservation of CvpA across diverse bacterial phyla suggests that this membrane protein facilitates cell envelope homeostasis in response to varied cell envelope perturbations.

Optical biosensors for bacteria detection by a peptidomimetic antimicrobial compound

The Analyst ◽  
2015 ◽  
Vol 140 (22) ◽  
pp. 7726-7733 ◽  
Author(s):  
Elena Tenenbaum ◽  
Ester Segal
Keyword(s):  
Bacteria Detection ◽  
Optical Biosensors ◽  
Label Free ◽  
Antimicrobial Compound ◽  
Porous Si ◽  
E Coli ◽  
Optical Transducer

A sensitive and label-free biosensor for E. coli detection, based on a peptidomimetic antimicrobial compound, which is tethered to a nanostructured porous Si optical transducer is presented.

Electrochemical Detection of Bacteria Using Graphene Oxide Electrodeposited on Titanium Implants

2014 ◽  
Vol 96 ◽  
pp. 45-53 ◽  
Author(s):  
Sirinrath Sirivisoot ◽  
Yardnapar Parcharoen ◽  
Thomas J. Webster
Keyword(s):  
Electron Transfer ◽  
Graphene Oxide ◽  
Electrochemical Sensors ◽  
Direct Electron Transfer ◽  
Orthopedic Implants ◽  
Titanium Implants ◽  
Cyclic Voltammograms ◽  
Label Free ◽  
Bacteria Growth ◽  
E Coli

Graphene oxide was electrodeposited on titanium (Ti-GO) and anodized titanium (ATi-GO) as label-free sensors for the detection of challenging living organisms, specificallyEscherichia coli(E. coli) andStaphylococcus aureus(S. aureus). The graphene modification contributed to two sets of oxidation-reduction peaks in cyclic voltammograms (CVs) of bacteria growth on the electrode surfaces (ATi-GO) that resulted in increasing direct electron transfer and stimulating excretion of mediating molecules for higher electron transfer between electrodes and bacteria. Additionally, similar wave patterns of CVs were found whenE. coliorS. aureuswere grown and electrocatalyzed on ATi-GO. The results suggest that bacteria on titanium implant surfaces could be easily detected by using mediatorless ATi-GO sensors electrochemically. These finding open another interesting method in using ATi-GO asin situelectrochemical sensors for label-free, close to real-time detection of bacteria infection in orthopedic implants.

Functionalized Polymers as Receptors for Detection of Cells

2011 ◽  
Vol 64 (9) ◽  
pp. 1256 ◽  
Author(s):  
Miroslava Polreichova ◽  
Usman Latif ◽  
Franz L. Dickert
Keyword(s):  
Soft Lithography ◽  
Quartz Crystal ◽  
Coli Strain ◽  
Yeast Cells ◽  
Label Free ◽  
Sensitive Material ◽  
E Coli ◽  
Label Free Detection ◽  
Quartz Crystal Microbalances ◽  
Cell Strains

Mass sensitive sensors were applied for fast and label-free detection of bio-analytes. Robust and miniaturized sensor devices were fabricated by combining bio-mimetic imprinted surfaces with quartz crystal microbalances for the analysis of yeast and bacteria cells. These sensors allow us to differentiate between different growing stages of yeast cells. Moreover, the viability of cells was detected by structuring quartz crystal microbalance electrodes like a grid. Artificial yeast cells were produced to pattern the recognition layer, giving reversible enrichment of the respective bio-analytes. This approach was followed to ensure the reproducibility of the identical sensitive material in each case, because the properties of each cell depend on its growth stage, which varies over time. The strategy was further applied to develop a sensitive system for Escherichia coli. Structuring of these materials by soft lithography allows differentiation between cell strains, e.g. E. coli (strain W & B) with a five-fold selectivity.

scholarly journals Identification of two fnr genes and characterisation of their role in the anaerobic switch in Sphingopyxis granuli strain TFA

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yolanda Elisabet González-Flores ◽  
Rubén de Dios ◽  
Francisca Reyes-Ramírez ◽  
Eduardo Santero
Keyword(s):  
Nitric Oxide ◽  
Energy Source ◽  
Stress Response ◽  
Organic Solvent ◽  
Double Mutant ◽  
Anaerobic Growth ◽  
Catabolic Pathway ◽  
E Coli ◽  
Stress Response Genes ◽  
Induced Stress

AbstractSphingopyxis granuli strain TFA is able to grow on the organic solvent tetralin as the only carbon and energy source. The aerobic catabolic pathway for tetralin, the genes involved and their regulation have been fully characterised. Unlike most of the bacteria belonging to the sphingomonads group, this strain is able to grow in anoxic conditions by respiring nitrate, though not nitrite, as the alternative electron acceptor. In this work, two fnr-like genes, fnrN and fixK, have been identified in strain TFA. Both genes are functional in E. coli and Sphingopyxis granuli although fixK, whose expression is apparently activated by FnrN, seems to be much less effective than fnrN in supporting anaerobic growth. Global transcriptomic analysis of a ΔfnrN ΔfixK double mutant and identification of Fnr boxes have defined a minimal Fnr regulon in this bacterium. However, expression of a substantial number of anaerobically regulated genes was not affected in the double mutant. Additional regulators such regBA, whose expression is also activated by Fnr, might also be involved in the anaerobic response. Anaerobically induced stress response genes were not regulated by Fnr but apparently induced by stress conditions inherent to anaerobic growth, probably due to accumulation of nitrite and nitric oxide.

Sign in / Sign up

Export Citation Format

Share Document