The Escherichia coli CitT transporter can be used as a succinate exporter for succinate production

2020 ◽  
Author(s):  
Sousuke Takahashi ◽  
Mayu Miyachi ◽  
Hisanori Tamaki ◽  
Hideyuki Suzuki
Keyword(s):  
Escherichia Coli ◽  
Culture Medium ◽  
Isocitrate Lyase ◽  
Aerobic Condition ◽  
Coli Strain ◽  
Succinate Production ◽  
E Coli ◽  
Citrate Transporter ◽  
Fed Batch Culture ◽  
Mg1655 Strain

ABSTRACT Escherichia coli strain, whose gene is one of the subunits of succinate dehydrogenase (sdhA), and gene of the transcriptional repressor of isocitrate lyase (iclR) were disrupted, accumulated 6.6 times as much intracellular succinate as the wild-type MG1655 strain in aerobic growth, but succinate was not found in the culture medium. E. coli citT gene that encodes a citrate transporter was cloned under the control of the lacI promoter in pBR322-based plasmid and the above strain was transformed. This transformant, grown under aerobic condition in M9-tryptone medium with citrate, accumulated succinate in the medium while no succinate was found in the medium without citrate. CitT was active as a succinate transporter for 168 h by changing the culture medium or for 24 h in fed-batch culture. This study suggests that the CitT transporter functions as a succinate exporter in E. coli for succinate production in the presence of citrate.

scholarly journals Regulation of Expression of the TIR-Containing Protein C Gene of the Uropathogenic Escherichia coli Strain CFT073

Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 549
Author(s):  
Julia Ittensohn ◽  
Jacqueline Hemberger ◽  
Hannah Griffiths ◽  
Maren Keller ◽  
Simone Albrecht ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein C ◽  
Interleukin 1 ◽  
Coli Strain ◽  
Uropathogenic Escherichia Coli ◽  
Reporter Construct ◽  
Regulation Of Expression ◽  
E Coli ◽  
Strain Cft073 ◽  
Myeloid Differentiation Factor

The uropathogenic Escherichia coli strain CFT073 causes kidney abscesses in mice Toll/interleukin-1 receptor domain-containing protein C (TcpC) dependently and the corresponding gene is present in around 40% of E. coli isolates of pyelonephritis patients. It impairs the Toll-like receptor (TLR) signaling chain and the NACHT leucin-rich repeat PYD protein 3 inflammasome (NLRP3) by binding to TLR4 and myeloid differentiation factor 88 as well as to NLRP3 and caspase-1, respectively. Overexpression of the tcpC gene stopped replication of CFT073. Overexpression of several tcpC-truncation constructs revealed a transmembrane region, while its TIR domain induced filamentous bacteria. Based on these observations, we hypothesized that tcpC expression is presumably tightly controlled. We tested two putative promoters designated P1 and P2 located at 5′ of the gene c2397 and 5′ of the tcpC gene (c2398), respectively, which may form an operon. High pH and increasing glucose concentrations stimulated a P2 reporter construct that was considerably stronger than a P1 reporter construct, while increasing FeSO4 concentrations suppressed their activity. Human urine activated P2, demonstrating that tcpC might be induced in the urinary tract of infected patients. We conclude that P2, consisting of a 240 bp region 5′ of the tcpC gene, represents the major regulator of tcpC expression.

scholarly journals Transcriptomic Analysis of Shiga Toxin-Producing Escherichia coli during Initial Contact with Cattle Colonic Explants

2020 ◽  
Vol 8 (11) ◽  
pp. 1662
Author(s):  
Zachary R. Stromberg ◽  
Rick E. Masonbrink ◽  
Melha Mellata
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Bacterial Colonization ◽  
Coli Strain ◽  
Fold Change ◽  
Initial Contact ◽  
Lon Protease ◽  
E Coli ◽  
Log2 Fold Change ◽  
Colonization Factors

Foodborne pathogens are a public health threat globally. Shiga toxin-producing Escherichia coli (STEC), particularly O26, O111, and O157 STEC, are often associated with foodborne illness in humans. To create effective preharvest interventions, it is critical to understand which factors STEC strains use to colonize the gastrointestinal tract of cattle, which serves as the reservoir for these pathogens. Several colonization factors are known, but little is understood about initial STEC colonization factors. Our objective was to identify these factors via contrasting gene expression between nonpathogenic E. coli and STEC. Colonic explants were inoculated with nonpathogenic E. coli strain MG1655 or STEC strains (O26, O111, or O157), bacterial colonization levels were determined, and RNA was isolated and sequenced. STEC strains adhered to colonic explants at numerically but not significantly higher levels compared to MG1655. After incubation with colonic explants, flagellin (fliC) was upregulated (log2 fold-change = 4.0, p < 0.0001) in O157 STEC, and collectively, Lon protease (lon) was upregulated (log2 fold-change = 3.6, p = 0.0009) in STEC strains compared to MG1655. These results demonstrate that H7 flagellum and Lon protease may play roles in early colonization and could be potential targets to reduce colonization in cattle.

scholarly journals Construction of an artificial biosynthetic pathway for hyperextended archaeal membrane lipids in the bacterium Escherichia coli

2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Ryo Yoshida ◽  
Hisashi Hemmi
Keyword(s):  
Escherichia Coli ◽  
Biosynthetic Pathway ◽  
Membrane Lipids ◽  
Extreme Environments ◽  
Halophilic Archaea ◽  
Coli Strain ◽  
Glycerol Moiety ◽  
Hyperthermophilic Archaeon ◽  
E Coli ◽  
Aeropyrum Pernix

Abstract Archaea produce unique membrane lipids, which possess two fully saturated isoprenoid chains linked to the glycerol moiety via ether bonds. The isoprenoid chain length of archaeal membrane lipids is believed to be important for some archaea to thrive in extreme environments because the hyperthermophilic archaeon Aeropyrum pernix and some halophilic archaea synthesize extended C25,C25-archaeal diether-type membrane lipids, which have isoprenoid chains that are longer than those of typical C20,C20-diether lipids. Natural archaeal diether lipids possessing longer C30 or C35 isoprenoid chains, however, have yet to be isolated. In the present study, we attempted to synthesize such hyperextended archaeal membrane lipids. We investigated the substrate preference of the enzyme sn-2,3-(digeranylfarnesyl)glycerol-1-phosphate synthase from A. pernix, which catalyzes the transfer of the second C25 isoprenoid chain to the glycerol moiety in the biosynthetic pathway of C25,C25-archaeal membrane lipids. The enzyme was shown to accept sn-3-hexaprenylglycerol-1-phosphate, which has a C30 isoprenoid chain, as a prenyl acceptor substrate to synthesize sn-2-geranylfarnesyl-3-hexaprenylglycerol-1-phosphate, a supposed precursor for hyperextended C25,C30-archaeal membrane lipids. Furthermore, we constructed an artificial biosynthetic pathway by introducing 4 archaeal genes and 1 gene from Bacillus subtilis in the cells of Escherichia coli, which enabled the E. coli strain to produce hyperextended C25,C30-archaeal membrane lipids, which have never been reported so far.

scholarly journals Evaluation of Physiological Effects Induced by Manuka Honey Upon Staphylococcus aureus and Escherichia coli

2019 ◽  
Vol 7 (8) ◽  
pp. 258 ◽  
Author(s):  
Patricia Combarros-Fuertes ◽  
Leticia M. Estevinho ◽  
Rita Teixeira-Santos ◽  
Acácio G. Rodrigues ◽  
Cidália Pina-Vaz ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Membrane Potential ◽  
Membrane Integrity ◽  
Efflux Pumps ◽  
Coli Strain ◽  
Antibacterial Action ◽  
Manuka Honey ◽  
E Coli ◽  
Action Mechanisms

Several studies have explored the antimicrobial properties of manuka honey (MkH). However, the data available regarding antibacterial action mechanisms are scarcer. The aim of this study was to scrutinize and characterize primary effects of manuka honey (MkH) upon the physiological status of Staphylococcus aureus and Escherichia coli (as Gram-positive and Gram-negative bacteria models, respectively), using flow cytometry (FC) to reveal its antibacterial action mechanisms. Effects of MkH on membrane potential, membrane integrity and metabolic activity were assessed using different fluorochromes in a 180 min time course assay. Time-kill experiments were carried out under the same conditions. Additionally, MkH effect on efflux pumps was also studied in an E. coli strain with an over-expression of several efflux pumps. Exposure of bacteria to MkH resulted in physiological changes related to membrane potential and membrane integrity; these effects displayed slight differences among bacteria. MkH induced a remarkable metabolic disruption as primary physiological effect upon S. aureus and was able to block efflux pump activity in a dose-dependent fashion in the E. coli strain.

scholarly journals Uropathogenic Escherichia coli Biofilm-Forming Capabilities are not Predictable from Clinical Details or from Colonial Morphology

Diseases ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 11 ◽  
Author(s):  
Shane Whelan ◽  
Mary Claire O’Grady ◽  
Dan Corcoran ◽  
Karen Finn ◽  
Brigid Lucey
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Culture Medium ◽  
Recurrent Infection ◽  
Positive Control ◽  
Uropathogenic Escherichia Coli ◽  
E Coli ◽  
Recurrent Uti ◽  
Colonial Morphology ◽  
The Difference

Antibiotic resistance is increasing to an extent where efficacy is not guaranteed when treating infection. Biofilm formation has been shown to complicate treatment, whereby the formation of biofilm is associated with higher minimum inhibitory concentration values of antibiotic. The objective of the current paper was to determine whether biofilm formation is variable among uropathogenic Escherichia coli isolates and whether formation is associated with recurrent urinary tract infection (UTI), and whether it can be predicted by phenotypic appearance on culture medium A total of 62 E. coli isolates that were reported as the causative agent of UTI were studied (33 from patients denoted as having recurrent UTI and 29 from patients not specified as having recurrent UTI). The biofilm forming capability was determined using a standard microtitre plate method, using E. coli ATCC 25922 as the positive control. The majority of isolates (93.6%) were found to be biofilm formers, whereby 81% were denoted as strong or very strong producers of biofilm when compared to the positive control. Through the use of a Wilcox test, the difference in biofilm forming propensity between the two patient populations was found to not be statistically significant (p = 0.5). Furthermore, it was noted that colony morphology was not a reliable predictor of biofilm-forming propensity. The findings of this study indicate that biofilm formation is very common among uropathogens, and they suggest that the biofilm-forming capability might be considered when treating UTI. Clinical details indicating a recurrent infection were not predictors of biofilm formation.

scholarly journals Complete Genome Sequence of Escherichia coli Strain WG5

2018 ◽  
Vol 6 (2) ◽  
Author(s):  
Lejla Imamovic ◽  
Maria-Anna Misiakou ◽  
Eric van der Helm ◽  
Gianni Panagiotou ◽  
Maite Muniesa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Coli Strain ◽  
Somatic Coliphages ◽  
E Coli

ABSTRACT Escherichia coli strain WG5 is a widely used host for phage detection, including somatic coliphages employed as standard ISO method 10705-1 (2000). Here, we present the complete genome sequence of a commercial E. coli WG5 strain.

THE CONVERSION OF 2,6-DIAMINOPIMELIC ACID-1,7-C14TO LYSINE-1-C14BY CERTAIN BACTERIA

1961 ◽  
Vol 39 (10) ◽  
pp. 1551-1558 ◽  
Author(s):  
A. J. Finlayson ◽  
F. J. Simpson
Keyword(s):  
Escherichia Coli ◽  
Aspergillus Flavus ◽  
Culture Medium ◽  
Leuconostoc Mesenteroides ◽  
Streptomyces Griseus ◽  
Diaminopimelic Acid ◽  
Proteus Vulgaris ◽  
E Coli ◽  
Carbon 14 ◽  
Carboxyl Carbon

When 2,6-diaminopimelicacid-1,7-C14was added to growing cultures of Bacillus megaterium, Staphlococcus aureus, and Escherichia coli, 8–9% of added carbon-14 appeared in the cellular lysine. Similar experiments with Proteus vulgaris, Streptomyces griseus, Aspergillus flavus, and Lactobacillus arabinosus resulted in less than 0.3% of the added carbon-14 being incorporated into the cellular lysine. Leuconostoc mesenteroides converted 0.6% of the added DAP-1,7-C14to lysine-1-C14.Over 90% of the carbon-14 in cell lysine from B. megaterium and L. mesenteroides was found in the carboxyl carbon. This was interpreted as indicating a direct decarboxylation of DAP-1,7-C14to lysine-1-C14. About 70% of the carbon-14 in the lysine from cells of S. aureus and E. coli was found in the carboxyl carbon, thus suggesting that some lysine comes from sources other than 2,6-diaminopimelic acid.Those organisms that actively decarboxylated DAP-1,7-C14to form lysine-C14also synthesized DAP and excreted it into the culture medium during growth.

scholarly journals Dynamic Mechanisms of the Bactericidal Action of an Al2O3-TiO2-Ag Granular Material on an Escherichia coli Strain

2015 ◽  
Vol 81 (20) ◽  
pp. 7135-7142 ◽  
Author(s):  
Marie-Anne Tartanson ◽  
Laurence Soussan ◽  
Matthieu Rivallin ◽  
Sophie Pecastaings ◽  
Cristian V. Chis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Granular Material ◽  
Morphological Changes ◽  
Membrane Damage ◽  
Coli Strain ◽  
Cell Integrity ◽  
Bactericidal Action ◽  
Intact Cells ◽  
Content Type ◽  
E Coli

ABSTRACTThe bactericidal activity of an Al2O3-TiO2-Ag granular material against anEscherichia colistrain was confirmed by a culture-based method. In particular, 100% of microorganisms were permanently inactivated in 30 to 45 min. The present work aimed to investigate the mechanisms of the bactericidal action of this material and their dynamics onEscherichia coliusing different techniques. Observations by transmission electron microscopy (TEM) at different times of disinfection revealed morphological changes in the bacteria as soon as they were put in contact with the material. Notably highlighted were cell membrane damage; cytoplasm detachment; formation of vacuoles, possibly due to DNA condensation, in association with regions exhibiting different levels of electron density; and membrane lysis. PCR and flow cytometry analyses were used to confirm and quantify the observations of cell integrity. The direct exposure of cells to silver, combined with the oxidative stress induced by the reactive oxygen species (ROS) generated, was identified to be responsible for these morphological alterations. From the first 5 min of treatment with the Al2O3-TiO2-Ag material, 98% ofE. coliisolates were lysed. From 30 min, cell viability decreased to reach total inactivation, although approximately 1% of permeableE. colicells and 1% of intact cells (105genomic units · ml−1) were evidenced. This study demonstrates that the bactericidal effect of the material results from a synergic action of desorbed and supported silver. Supported silver was shown to generate the ROS evidenced.

scholarly journals Preparation of gram quantities of a purified R-factor-mediated penicillinase from Escherichia coli strain W3310

1972 ◽  
Vol 130 (1) ◽  
pp. 55-62 ◽  
Author(s):  
J. Melling ◽  
G. K. Scott
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Enzyme Activity ◽  
Coli Strain ◽  
Specific Enzyme ◽  
Specific Enzyme Activity ◽  
E Coli ◽  
R Factor

Purified penicillinase, in gram quantities, has been prepared from Escherichia coli strain W3310 by using methods developed to handle large amounts of material. The final product had a specific enzyme activity of 3.08 units/μg of protein, which was over twice as high as that reported previously (Datta & Richmond, 1966). The purified enzyme was similar to that from E. coli strain TEM, but different in molecular weight and some other respects. The differences observed may be a result of the greater purity obtained.

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