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Proteomics-based mechanistic investigation of Escherichia coli inactivation by pulsed electric field

ABSTRACT: mechanistic investigation of Escherichia coli inactivation

ORGANISM(S): Escherichia Coli Vdp

SUBMITTER: Yanbo Song

PROVIDER: PXD014365 | iProX | Tue Jun 25 00:00:00 BST 2019

REPOSITORIES: iProX

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Proteomics-Based Mechanistic Investigation of <i>Escherichia coli</i> Inactivation by Pulsed Electric Field.

Liu Zhenyu Z Zhao Lingying L Zhang Qin Q Huo Nan N Shi Xiaojing X Li Linwei L Jia Liyan L Lu Yuanyuan Y Peng Yong Y Song Yanbo Y

Frontiers in microbiology 20191108

The pulsed electric field (PEF) technology has been widely applied to inactivate pathogenic bacteria in food products. Though irreversible pore formation and membrane disruption is considered to be the main contributing factor to PEF's sterilizing effects, the exact molecular mechanisms remain poorly understood. In this study, by using mass spectrometry (MS)-based label-free quantitative proteomic analysis, we compared the protein profiles of PEF-treated and untreated <i>Escherichia coli</i>. We ...[more]

PMID: 31781086

Similar Datasets

Proteomics-Based Mechanistic Investigation of Escherichia coli Inactivation by Pulsed Electric Field.

Project description:The pulsed electric field (PEF) technology has been widely applied to inactivate pathogenic bacteria in food products. Though irreversible pore formation and membrane disruption is considered to be the main contributing factor to PEF's sterilizing effects, the exact molecular mechanisms remain poorly understood. In this study, by using mass spectrometry (MS)-based label-free quantitative proteomic analysis, we compared the protein profiles of PEF-treated and untreated Escherichia coli. We identified a total of 175 differentially expressed proteins, including 52 candidates that were only detected in at least two of the three samples in one experiment group but not in the other group. Functional analysis revealed that the differential proteins were primarily involved in the regulation of cell membrane composition and integrity, stress response, as well as various metabolic processes. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis was conducted on the genes of selected differential proteins at varying PEF intensities, which were known to result in different cell killing levels. The qRT-PCR data confirmed that the proteomic results could be reliably used for further data interpretation, and that the changes in the expression levels of the differential candidates were, to a large extent, caused directly by the PEF treatment. The findings of the current study offered valuable insight into PEF-induced cell inactivation.

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Proteomics-based mechanistic investigation of Staphylococcus aureus inactivation by pulsed electric field

Project description:The Staphylococcus aureus strain was cultured in LB medium at 150 rpm and 37 ℃ to a final optical density (OD) of 1.0 at 600 n m. Subsequently, 1 m L of the culture broth was collected and centrifuged at 8000 rpm, 4 ℃ for 2 min. The cell pellet was re-suspended in 2 m L of ice-cold 0.1% (w/v) bacteriological peptone solution and divided into 70 µL aliquots. For PEF treatment, each aliquot was pipetted into a sterile 1.0 mm electroporation cuvette and pulsed using a BTX ECM830 Square Wave Electroporation System (BTX, United States). For a cell killing extent of 94.89±0.90% , pulse intensity,number and duration were set to 11 kV·cm-1 ,86 and 70μs，respectively. The killing extent was 50.29±5.27% with the treatment parameters of 9kV·cm-1 , pulse number of 50, pulse duration of 50μs. For controls, a mock treatment without the application of PEF was performed, with identical downstream procedures. The whole experiment described above was conducted in triplicate. Extract the total protein of the sample and perform quality inspection, reductive alkylation and enzymatic hydrolysis, TMT labeling and mixing, and perform two-dimensional analysis by RPLC one-dimensional separation and liquid phase tandem mass spectrometry (Easy-nLC 1200 combined with Q Exactive mass spectrometer). The raw files off the computer are analyzed by ProteomeDiscovererTM Software 2.2, and the searched species is the self-tested Staphylococcus aureus CGMCCC26003 genome database.

2021-06-17 | PXD026761 |

Transcriptional profile of pulsed AsxR in Escherichia coli O157:H7 str. TUV93-0

Project description:Transcript abundance in Escherichia coli O157:H7 was determined in the presence or absence of pulsed expression of the small RNA, AsxR.

2014-05-22 | GSE46113 | GEO

Psychological Evaluation According Colonic Colonization by Escherichia Coli of Patients After Colorectal Surgery

Project description:The purpose of this study is to determine whether the presence of pathogenic Escherichia coli in colon is associated with psychiatric disorders.

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Transcriptional profile of pulsed AsxR in Escherichia coli O157:H7 str. TUV93-0

Project description:Transcript abundance in Escherichia coli O157:H7 was determined in the presence or absence of pulsed expression of the small RNA, AsxR. AsxR was cloned under the control the arabinose inducible promoter Para. Escherichia coli O157:H7 str. TUV93-0 with pAsxR or empty vector was cultured in MEM-HEPES media to an OD600 of 0.8 and 0.2% arabinose added. 10min after addition of arabinose 10ml of cells were harvested and and pellets resuspended in 1ml of Trizol and total RNA isolated. RNAs were labelled using the SuperScript Plus indirect cDNA labelling System. Triplicate control RNAs were pooled and hybridised to seperate AsxR test RNAs on three microarays. Arrays were hybridised using the Maui hybridisation platform and Scann using and Axon Autoloader Scanner. GenePix software was used to analyse images and GPR files were analysed using Genespring 7.3.1.

2014-05-22 | E-GEOD-46113 | biostudies-arrayexpress

Microaerobic fermentation alters lactose metabolism in Escherichia coli

Project description:We report the effect of oxygenation state in lactose grown escherichia coli producing recombinant proteins. To shed more light on the mechanistic correlation between the uptake of lactose and dissolved oxygen, a comprehensive study has been undertaken with the E. coli BL21 (DE3) strain. Differences in consumption pattern of lactose, metabolites, biomass and product formation due to aerobiosis have been investigated. Transcriptomic profiling of metabolic changes due to aerobic process and microaerobic process during protein formation phase has been studied and the results provide a deeper understanding of protein production in E. coli BL21 (DE3) strains with lactose based promoter expression systems.This study also provides a scientific understanding of escherichia coli metabolism upon oxygen fluctuations.

2020-05-18 | GSE135698 | GEO

600-ns pulsed electric fields affect inactivation and antibiotic susceptibilities of Escherichia coli and Lactobacillus acidophilus.

Project description:Cell suspensions of Escherichia coli and Lactobacillus acidophilus were exposed to 600-ns pulsed electric fields (nsPEFs) at varying amplitudes (Low-13.5, Mid-18.5 or High-23.5 kV cm-1) and pulse numbers (0 (sham), 1, 5, 10, 100 or 1000) at a 1 hertz (Hz) repetition rate. The induced temperature rise generated at these exposure parameters, hereafter termed thermal gradient, was measured and applied independently to cell suspensions in order to differentiate inactivation triggered by electric field (E-field) from heating. Treated cell suspensions were plated and cellular inactivation was quantified by colony counts after a 24-hour (h) incubation period. Additionally, cells from both exposure conditions were incubated with various antibiotic-soaked discs to determine if nsPEF exposure would induce changes in antibiotic susceptibility. Results indicate that, for both species, the total delivered energy (amplitude, pulse number and pulse duration) determined the magnitude of cell inactivation. Specifically, for 18.5 and 23.5 kV cm-1 exposures, L. acidophilus was more sensitive to the inactivation effects of nsPEF than E. coli, however, for the 13.5 kV cm-1 exposures E. coli was more sensitive, suggesting that L. acidophilus may need to meet an E-field threshold before significant inactivation can occur. Results also indicate that antibiotic susceptibility was enhanced by multiple nsPEF exposures, as observed by increased zones of growth inhibition. Moreover, for both species, a temperature increase of ≤ 20 °C (89% of exposures) was not sufficient to significantly alter cell inactivation, whereas none of the thermal equivalent exposures were sufficient to change antibiotic susceptibility categories.

| S-EPMC7080936 | biostudies-literature

Gene expression from Escherichia coli.

Project description:Gene expression from Escherichia coli.

2013-07-12 | GSE48776 | GEO

Investigation on the anaerobic propionate degradation by Escherichia coli K12

Project description:Propionate is an abundant carboxylic acid in nature. Microorganisms metabolize propionate aerobically via the 2-methylcitrate pathway. This pathway depends on a series of three reactions in the citric acid cycle that leads to the conversion of succinate to oxaloacetate. Interestingly, the gamma-proteobacterium Escherichia coli can use propionate as a carbon and electron source under oxic but not under anoxic conditions. The typical downregulation of the citric acid cycle under anoxic conditions is only partially responsible for the inability to use propionate under anoxic conditions since an arcA mutant shows very limited growth on propionate. RT-PCR and transcriptomic analysis revealed a post-transcriptional regulation of the prp-genecluster encoding the necessary enzymes for propionate metabolism. The polycistronic mRNA was hydrolyzed in the 3`-5` direction under anoxic conditions. This regulatory strategy is highly constructive because the last gene of the operon encodes the first enzyme of the propionate metabolism. Further analysis revealed that RNase R catalyzes the hydrolysis of the prp transcripts. Consequently, an rnr-deletion strain could metabolize propionate under anoxic conditions. To the best of our knowledge, this is the first study describing the influence of RNase R on the anaerobic metabolism of E. coli.

2017-12-14 | GSE78814 | GEO

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