Quantitative Assessment of Oxygen Availability: Perceived Aerobiosis and Its Effect on Flux Distribution in the Respiratory Chain of Escherichia coli

ABSTRACT Despite a large number of studies on the role of oxygen in cellular processes, there is no consensus as to how oxygen availability to the cell should be defined, let alone how it should be quantified. Here, a quantitative definition for oxygen availability (perceived aerobiosis) is presented; the definition is based on a calibration with reference to the minimal oxygen supply rate needed for fully oxidative catabolism (i.e., complete conversion of the energy source to CO2 and water for glucose-limited conditions). This quantitative method is used to show how steady-state electron fluxes through the alternative cytochrome oxidases of Escherichia coli are distributed as a function of the extent of aerobiosis of glucose-limited chemostat cultures. At low oxygen availability the electron flux is mainly via the high-affinity cytochrome bd oxidase, and, at higher oxygen availability, a similar phenomenon occurs but now via the low-affinity cytochrome bo oxidase. The main finding is that the catabolic activities of E. coli (and specifically its respiratory activity) are affected by the actual oxygen availability per unit of biomass rather than by the residual dissolved oxygen concentration of the culture.

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Mapping Stress-Induced Changes in Autoinducer AI-2 Production in Chemostat-Cultivated Escherichia coli K-12

Journal of Bacteriology ◽

10.1128/jb.183.9.2918-2928.2001 ◽

2001 ◽

Vol 183 (9) ◽

pp. 2918-2928 ◽

Cited By ~ 71

Author(s):

Matthew P. DeLisa ◽

James J. Valdes ◽

William E. Bentley

Keyword(s):

Escherichia Coli ◽

Steady State ◽

Quorum Sensing ◽

Interleukin 2 ◽

Heterologous Proteins ◽

Induced Expression ◽

E Coli ◽

Chemostat Cultures ◽

K 12

ABSTRACT Numerous gram-negative bacteria employ a cell-to-cell signaling mechanism, termed quorum sensing, for controlling gene expression in response to population density. Recently, this phenomenon has been discovered in Escherichia coli, and while pathogenicE. coli utilize quorum sensing to regulate pathogenesis (i.e., expression of virulence genes), the role of quorum sensing in nonpathogenic E. coli is less clear, and in particular, there is no information regarding the role of quorum sensing during the overexpression of recombinant proteins. The production of autoinducer AI-2, a signaling molecule employed by E. coli for intercellular communication, was studied in E. coli W3110 chemostat cultures using a Vibrio harveyi AI-2 reporter assay (M. G. Surrette and B. L. Bassler, Proc. Natl. Acad. Sci. USA 95:7046–7050, 1998). Chemostat cultures enabled a study of AI-2 regulation through steady-state and transient responses to a variety of environmental stimuli. Results demonstrated that AI-2 levels increased with the steady-state culture growth rate. In addition, AI-2 increased following pulsed addition of glucose, Fe(III), NaCl, and dithiothreitol and decreased following aerobiosis, amino acid starvation, and isopropyl-β-d-thiogalactopyranoside-induced expression of human interleukin-2 (hIL-2). In general, the AI-2 responses to several perturbations were indicative of a shift in metabolic activity or state of the cells induced by the individual stress. Because of our interest in the expression of heterologous proteins in E. coli, the transcription of four quorum-regulated genes and 20 stress genes was mapped during the transient response to induced expression of hIL-2. Significant regulatory overlap was revealed among several stress and starvation genes and known quorum-sensing genes.

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Roles of the C-Terminal Amino Acids of Non-Hexameric Helicases: Insights from Escherichia coli UvrD

International Journal of Molecular Sciences ◽

10.3390/ijms22031018 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1018

Author(s):

Hiroaki Yokota

Keyword(s):

Escherichia Coli ◽

Amino Acids ◽

Amino Acid ◽

Single Molecule ◽

Underlying Mechanism ◽

Amino Acid Sequences ◽

E Coli ◽

X Ray Crystallography ◽

Terminal Amino

Helicases are nucleic acid-unwinding enzymes that are involved in the maintenance of genome integrity. Several parts of the amino acid sequences of helicases are very similar, and these quite well-conserved amino acid sequences are termed “helicase motifs”. Previous studies by X-ray crystallography and single-molecule measurements have suggested a common underlying mechanism for their function. These studies indicate the role of the helicase motifs in unwinding nucleic acids. In contrast, the sequence and length of the C-terminal amino acids of helicases are highly variable. In this paper, I review past and recent studies that proposed helicase mechanisms and studies that investigated the roles of the C-terminal amino acids on helicase and dimerization activities, primarily on the non-hexermeric Escherichia coli (E. coli) UvrD helicase. Then, I center on my recent study of single-molecule direct visualization of a UvrD mutant lacking the C-terminal 40 amino acids (UvrDΔ40C) used in studies proposing the monomer helicase model. The study demonstrated that multiple UvrDΔ40C molecules jointly participated in DNA unwinding, presumably by forming an oligomer. Thus, the single-molecule observation addressed how the C-terminal amino acids affect the number of helicases bound to DNA, oligomerization, and unwinding activity, which can be applied to other helicases.

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Distribution of Lactoferrin Is Related with Dynamics of Neutrophils in Bacterial Infected Mice Intestine

Molecules ◽

10.3390/molecules25071496 ◽

2020 ◽

Vol 25 (7) ◽

pp. 1496 ◽

Cited By ~ 1

Author(s):

Li Liang ◽

Zhen-Jie Wang ◽

Guang Ye ◽

Xue-You Tang ◽

Yuan-Yuan Zhang ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Activity ◽

Mucosal Immunity ◽

Mrna Levels ◽

Iron Binding ◽

E Coli ◽

New Knowledge ◽

Activated Neutrophils ◽

Binding Glycoprotein

Lactoferrin (Lf) is a conserved iron-binding glycoprotein with antimicrobial activity, which is present in secretions that recover mucosal sites regarded as portals of invaded pathogens. Although numerous studies have focused on exogenous Lf, little is known about its expression of endogenous Lf upon bacterial infection. In this study, we investigated the distribution of Lf in mice intestine during Escherichia coli (E. coli) K88 infection. PCR and immunohistology staining showed that mRNA levels of Lf significantly increased in duodenum, ileum and colon, but extremely decreased in jejunum at 8 h and 24 h after infection. Meanwhile, endogenous Lf was mostly located in the lamina propria of intestine villi, while Lf receptor (LfR) was in the crypts. It suggested that endogenous Lf-LfR interaction might not be implicated in the antibacterial process. In addition, it was interesting to find that the infiltration of neutrophils into intestine tissues was changed similarly to Lf expression. It indicated that the variations of Lf expression were rather due to an equilibrium between the recruitment of neutrophils and degranulation of activated neutrophils. Thus, this new knowledge will pave the way to a more effective understanding of the role of Lf in intestinal mucosal immunity.

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SATP (YaaH), a succinate–acetate transporter protein in Escherichia coli

Biochemical Journal ◽

10.1042/bj20130412 ◽

2013 ◽

Vol 454 (3) ◽

pp. 585-595 ◽

Cited By ~ 45

Author(s):

Joana Sá-Pessoa ◽

Sandra Paiva ◽

David Ribas ◽

Inês Jesus Silva ◽

Sandra Cristina Viegas ◽

...

Keyword(s):

Escherichia Coli ◽

Acetic Acid ◽

Succinic Acid ◽

Critical Role ◽

Amino Acid Residues ◽

E Coli ◽

Transporter Protein ◽

Affinity Constants ◽

In Trans

In the present paper we describe a new carboxylic acid transporter in Escherichia coli encoded by the gene yaaH. In contrast to what had been described for other YaaH family members, the E. coli transporter is highly specific for acetic acid (a monocarboxylate) and for succinic acid (a dicarboxylate), with affinity constants at pH 6.0 of 1.24±0.13 mM for acetic acid and 1.18±0.10 mM for succinic acid. In glucose-grown cells the ΔyaaH mutant is compromised for the uptake of both labelled acetic and succinic acids. YaaH, together with ActP, described previously as an acetate transporter, affect the use of acetic acid as sole carbon and energy source. Both genes have to be deleted simultaneously to abolish acetate transport. The uptake of acetate and succinate was restored when yaaH was expressed in trans in ΔyaaH ΔactP cells. We also demonstrate the critical role of YaaH amino acid residues Leu131 and Ala164 on the enhanced ability to transport lactate. Owing to its functional role in acetate and succinate uptake we propose its assignment as SatP: the Succinate–Acetate Transporter Protein.

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Molecular survey of mcr1 and mcr2 plasmid mediated colistin resistance genes in Escherichia coli isolates of animal origin in Iran

BMC Research Notes ◽

10.1186/s13104-021-05519-6 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Kayhan Ilbeigi ◽

Mahdi Askari Badouei ◽

Hossein Vaezi ◽

Hassan Zaheri ◽

Sina Aghasharif ◽

...

Keyword(s):

Escherichia Coli ◽

Resistance Genes ◽

Companion Animals ◽

Multidrug Resistant ◽

Animal Origin ◽

Colistin Resistance ◽

E Coli ◽

High Level ◽

Farming Industry

Abstract Objectives The emergence of colistin-resistant Enterobacteriaceae from human and animal sources is one of the major public health concerns as colistin is the last-resort antibiotic for treating infections caused by multidrug-resistant Gram-negative bacteria. We aimed to determine the prevalence of the prototype widespread colistin resistance genes (mcr-1 and mcr-2) among commensal and pathogenic Escherichia coli strains isolated from food-producing and companion animals in Iran. Results A total of 607 E. coli isolates which were previously collected from different animal sources between 2008 and 2016 used to uncover the possible presence of plasmid-mediated colistin resistance genes (mcr-1 and mcr-2) by PCR. Overall, our results could not confirm the presence of any mcr-1 or mcr-2 positive E. coli among the studied isolates. It is concluded that despite the important role of food-producing animals in transferring the antibiotic resistance, they were not the main source for carriage of mcr-1 and mcr-2 in Iran until 2016. This study suggests that the other mcr variants (mcr-3 to mcr-9) might be responsible for conferring colistin resistance in animal isolates in Iran. The possible linkage between pig farming industry and high level of mcr carriage in some countries needs to be clarified in future prospective studies.

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Drinking water and diarrhoeal disease due to Escherichia coli

Journal of Water and Health ◽

10.2166/wh.2003.0008 ◽

2003 ◽

Vol 1 (2) ◽

pp. 65-72 ◽

Cited By ~ 49

Author(s):

Paul R. Hunter

Keyword(s):

Escherichia Coli ◽

Drinking Water ◽

Clinical Presentation ◽

Diarrhoeal Disease ◽

Central Place ◽

E Coli ◽

Related Disease ◽

Direct Damage ◽

Water Microbiology

Escherichia coli has had a central place in water microbiology for decades as an indicator of faecal pollution. It is only relatively recently that the role of E. coli as pathogen, rather than indicator, in drinking water has begun to be stressed. Interest in the role of E. coli as a cause of diarrhoeal disease has increased because of the emergence of E. coli O157:H7 and other enterohaemorrhagic E. coli, due to the severity of the related disease. There are enterotoxigenic, enteropathogenic, enterohaemorrhagic, enteroinvasive, enteroaggregative and diffusely adherent strains of E. coli. Each type of E. coli causes diarrhoeal disease through different mechanisms and each causes a different clinical presentation. Several of the types cause diarrhoea by the elaboration of one or more toxins, others by some other form of direct damage to epithelial cells. This paper discusses each of these types in turn and also describes their epidemiology, with particular reference to whether they are waterborne or not.

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Quantitative Determination of the Role of Lettuce Leaf Structures in Protecting Escherichia coli O157:H7 from Chlorine Disinfection

Journal of Food Protection ◽

10.4315/0362-028x-64.2.147 ◽

2001 ◽

Vol 64 (2) ◽

pp. 147-151 ◽

Cited By ~ 89

Author(s):

KAZUE TAKEUCHI ◽

JOSEPH F. FRANK

Keyword(s):

Escherichia Coli ◽

Leaf Surface ◽

Dead Cell ◽

Escherichia Coli O157 ◽

E Coli ◽

Chlorine Disinfection ◽

Sytox Green ◽

Tissue Surface ◽

Confocal Scanning

Viability of Escherichia coli O157:H7 cells on lettuce leaves after 200 mg/liter (200 ppm) chlorine treatment and the role of lettuce leaf structures in protecting cells from chlorine inactivation were evaluated by confocal scanning microscopy (CSLM). Lettuce samples (2 by 2 cm) were inoculated by immersing in a suspension containing 109 CFU/ml of E. coli O157: H7 for 24 ± 1 h at 4°C. Rinsed samples were treated with 200 mg/liter (200 ppm) chlorine for 5 min at 22°C. Viability of E. coli O157:H7 cells was evaluated by CSLM observation of samples stained with Sytox green (dead cell stain) and Alexa 594 conjugated antibody against E. coli O157:H7. Quantitative microscopic observations of viability were made at intact leaf surface, stomata, and damaged tissue. Most E. coli O157:H7 cells (68.3 ± 16.2%) that had penetrated 30 to 40 μm from the damaged tissue surface remained viable after chlorine treatment. Cells on the surface survived least (25.2 ± 15.8% survival), while cells that penetrated 0 to 10 μm from the damaged tissue surface or entered stomata showed intermediate survival (50.8 ± 13.5 and 45.6 ± 9.7% survival, respectively). Viability was associated with the depth at which E. coli O157:H7 cells were in the stomata. Although cells on the leaf surface were mostly inactivated, some viable cells were observed in cracks of cuticle and on the trichome. These results demonstrate the importance of lettuce leaf structures in the protection of E. coli O157:H7 cells from chlorine inactivation.

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Detection and Quantification of Superoxide Formed within the Periplasm of Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.00554-06 ◽

2006 ◽

Vol 188 (17) ◽

pp. 6326-6334 ◽

Cited By ~ 111

Author(s):

Sergei Korshunov ◽

James A. Imlay

Keyword(s):

Escherichia Coli ◽

Pathogenic Bacteria ◽

Phagocytic Cells ◽

Free Living ◽

E Coli ◽

Superoxide Formation ◽

Genes Encoding ◽

Copper Zinc ◽

Primary Substrate

ABSTRACT Many gram-negative bacteria harbor a copper/zinc-containing superoxide dismutase (CuZnSOD) in their periplasms. In pathogenic bacteria, one role of this enzyme may be to protect periplasmic biomolecules from superoxide that is released by host phagocytic cells. However, the enzyme is also present in many nonpathogens and/or free-living bacteria, including Escherichia coli. In this study we were able to detect superoxide being released into the medium from growing cultures of E. coli. Exponential-phase cells do not normally synthesize CuZnSOD, which is specifically induced in stationary phase. However, the engineered expression of CuZnSOD in growing cells eliminated superoxide release, confirming that this superoxide was formed within the periplasm. The rate of periplasmic superoxide production was surprisingly high and approximated the estimated rate of cytoplasmic superoxide formation when both were normalized to the volume of the compartment. The rate increased in proportion to oxygen concentration, suggesting that the superoxide is generated by the adventitious oxidation of an electron carrier. Mutations that eliminated menaquinone synthesis eradicated the superoxide formation, while mutations in genes encoding respiratory complexes affected it only insofar as they are likely to affect the redox state of menaquinone. We infer that the adventitious autoxidation of dihydromenaquinone in the cytoplasmic membrane releases a steady flux of superoxide into the periplasm of E. coli. This endogenous superoxide may create oxidative stress in that compartment and be a primary substrate of CuZnSOD.

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Molecular Characterization of Commensal Escherichia coli Adapted to Different Compartments of the Porcine Gastrointestinal Tract

Applied and Environmental Microbiology ◽

10.1128/aem.01688-12 ◽

2012 ◽

Vol 78 (19) ◽

pp. 6799-6803 ◽

Cited By ~ 12

Author(s):

Sam Abraham ◽

David M. Gordon ◽

James Chin ◽

Huub J. M. Brouwers ◽

Peter Njuguna ◽

...

Keyword(s):

Escherichia Coli ◽

Gastrointestinal Tract ◽

Random Amplified Polymorphic Dna ◽

Content Type ◽

E Coli ◽

Plasmid Replicon ◽

Different Strains ◽

Different Characteristics

ABSTRACTThe role ofEscherichia colias a pathogen has been the focus of considerable study, while much less is known about it as a commensal and how it adapts to and colonizes different environmental niches within the mammalian gut. In this study, we characterizeEscherichia coliorganisms (n= 146) isolated from different regions of the intestinal tracts of eight pigs (dueodenum, ileum, colon, and feces). The isolates were typed using the method of random amplified polymorphic DNA (RAPD) and screened for the presence of bacteriocin genes and plasmid replicon types. Molecular analysis of variance using the RAPD data showed thatE. coliisolates are nonrandomly distributed among different gut regions, and that gut region accounted for 25% (P< 0.001) of the observed variation among strains. Bacteriocin screening revealed that a bacteriocin gene was detected in 45% of the isolates, with 43% carrying colicin genes and 3% carrying microcin genes. Of the bacteriocins observed (H47, E3, E1, E2, E7, Ia/Ib, and B/M), the frequency with which they were detected varied with respect to gut region for the colicins E2, E7, Ia/Ib, and B/M. The plasmid replicon typing gave rise to 25 profiles from the 13 Inc types detected. Inc F types were detected most frequently, followed by Inc HI1 and N types. Of the Inc types detected, 7 were nonrandomly distributed among isolates from the different regions of the gut. The results of this study indicate that not only may the different regions of the gastrointestinal tract harbor different strains ofE. colibut also that strains from different regions have different characteristics.

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Aerobic Fermentation of d-Glucose by an Evolved Cytochrome Oxidase-Deficient Escherichia coli Strain

Applied and Environmental Microbiology ◽

10.1128/aem.00880-08 ◽

2008 ◽

Vol 74 (24) ◽

pp. 7561-7569 ◽

Cited By ~ 41

Author(s):

Vasiliy A. Portnoy ◽

Markus J. Herrgård ◽

Bernhard Ø. Palsson

Keyword(s):

Escherichia Coli ◽

Oxygen Uptake ◽

Growth Conditions ◽

Aerobic Growth ◽

Acid Fermentation ◽

Mixed Acid Fermentation ◽

E Coli ◽

Knockout Strain ◽

Mixed Acid ◽

Cytochrome Oxidases

ABSTRACT Fermentation of glucose to d-lactic acid under aerobic growth conditions by an evolved Escherichia coli mutant deficient in three terminal oxidases is reported in this work. Cytochrome oxidases (cydAB, cyoABCD, and cbdAB) were removed from the E. coli K12 MG1655 genome, resulting in the ECOM3 (E. coli cytochrome oxidase mutant) strain. Removal of cytochrome oxidases reduced the oxygen uptake rate of the knockout strain by nearly 85%. Moreover, the knockout strain was initially incapable of growing on M9 minimal medium. After the ECOM3 strain was subjected to adaptive evolution on glucose M9 medium for 60 days, a growth rate equivalent to that of anaerobic wild-type E. coli was achieved. Our findings demonstrate that three independently adaptively evolved ECOM3 populations acquired different phenotypes: one produced lactate as a sole fermentation product, while the other two strains exhibited a mixed-acid fermentation under oxic growth conditions with lactate remaining as the major product. The homofermenting strain showed a d-lactate yield of 0.8 g/g from glucose. Gene expression and in silico model-based analyses were employed to identify perturbed pathways and explain phenotypic behavior. Significant upregulation of ygiN and sodAB explains the remaining oxygen uptake that was observed in evolved ECOM3 strains. E. coli strains produced in this study showed the ability to produce lactate as a fermentation product from glucose and to undergo mixed-acid fermentation during aerobic growth.

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