The effect of proflavine on pyruvate kinase I of Escherichia coli B

1977 ◽  
Vol 55 (6) ◽  
pp. 618-624 ◽  
Author(s):  
Robin C. McKellar ◽  
Donn J. Kushner
Keyword(s):  
Escherichia Coli ◽  
Pyruvate Kinase ◽  
Hydrophobic Interactions ◽  
Glycolytic Enzyme ◽  
Molecular Probe ◽  
Inhibitory Effects ◽  
E Coli ◽  
The Hill ◽  
Escherichia Coli B ◽  
Inhibition Curve

Proflavine (PF) inhibited glucose use in sensitive but not resistant Escherichia coli B. Glucose transport (as measured by α-methylglucoside accumulation) was only partly inhibited by PF concentrations that completely blocked glucose use. Fructose 1,6-diphosphate-(FDP)-regulated pyruvate kinase (PK I) (EC 2.7.1.40), the only glycolytic enzyme affected by PF, was completely inhibited by a dye concentration of 0.8 mM. The inhibition curve for PF was sigmoidal, suggesting that PF was acting as an allosteric inhibitor. PF increased the K1/2 for phosphoenolpyruvate (PEP) and lowered the V; however, it had no effect on the Hill number for PEP. PF inhibition was partially reversed by FDP but not by cyclic AMP, AMP, ATP, fructose 6-phosphate, or dithiothreitol. Studies with a variety of acridines indicated that those substituted at the 3-position are the most effective inhibitors and also that hydrophobic interactions may be involved in PF inhibition of PK I. PK I for E. coli B/Pr was also strongly inhibited by PF, indicating that PF resistance does not lie at the level of this enzyme. Ribose-5-phosphate-regulated pyruvate kinase (EC 2.7.1.40) was much less sensitive than PK I to the inhibitory effects of PF. A role for PF as a molecular probe for PK I has been proposed.

Sites of Adsorption of the Bacteriophages T3 and T5 on the Wall of Escherichia Coli B.

1967 ◽  
Vol 25 ◽  
pp. 96-97
Author(s):  
Manfred E. Bayer
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Electron Microscope ◽  
Uranyl Acetate ◽  
E Coli ◽  
Receptor Sites ◽  
Rigid Cell Wall ◽  
Host Cell Surface ◽  
Bacterial Viruses ◽  
Escherichia Coli B

Bacterial viruses adsorb specifically to receptors on the host cell surface. Although the chemical composition of some of the cell wall receptors for bacteriophages of the T-series has been described and the number of receptor sites has been estimated to be 150 to 300 per E. coli cell, the localization of the sites on the bacterial wall has been unknown.When logarithmically growing cells of E. coli are transferred into a medium containing 20% sucrose, the cells plasmolize: the protoplast shrinks and becomes separated from the somewhat rigid cell wall. When these cells are fixed in 8% Formaldehyde, post-fixed in OsO4/uranyl acetate, embedded in Vestopal W, then cut in an ultramicrotome and observed with the electron microscope, the separation of protoplast and wall becomes clearly visible, (Fig. 1, 2). At a number of locations however, the protoplasmic membrane adheres to the wall even under the considerable pull of the shrinking protoplast. Thus numerous connecting bridges are maintained between protoplast and cell wall. Estimations of the total number of such wall/membrane associations yield a number of about 300 per cell.

Screening of Commercial and Pecan Shell–Extracted Liquid Smoke Agents as Natural Antimicrobials against Foodborne Pathogens

2012 ◽  
Vol 75 (6) ◽  
pp. 1148-1152 ◽  
Author(s):  
ELLEN J. VAN LOO ◽  
D. BABU ◽  
PHILIP G. CRANDALL ◽  
STEVEN C. RICKE
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Salmonella Enteritidis ◽  
Antioxidant Properties ◽  
Inhibitory Effects ◽  
Natural Antimicrobials ◽  
E Coli ◽  
Liquid Smoke ◽  
Water Based ◽  
Smoke Sample

Liquid smoke extracts have traditionally been used as flavoring agents, are known to possess antioxidant properties, and serve as natural alternatives to conventional antimicrobials. The antimicrobial efficacies of commercial liquid smoke samples may vary depending on their source and composition and the methods used to extract and concentrate the smoke. We investigated the MICs of eight commercial liquid smoke samples against Salmonella Enteritidis, Staphylococcus aureus, and Escherichia coli. The commercial liquid smoke samples purchased were supplied by the manufacturer as water-based or concentrated extracts of smoke from different wood sources. The MICs of the commercial smokes to inhibit the growth of foodborne pathogens ranged from 0.5 to 6.0% for E. coli, 0.5 to 8.0% for Salmonella, and 0.38 to 6% for S. aureus. The MIC for each liquid smoke sample was similar in its effect on both E. coli and Salmonella. Solvent-extracted antimicrobials prepared using pecan shells displayed significant differences between their inhibitory concentrations depending on the type of solvent used for extraction. The results indicated that the liquid smoke samples tested in this study could serve as effective natural antimicrobials and that their inhibitory effects depended more on the solvents used for extraction than the wood source.

scholarly journals Modification by an R determinant for streptomycin of hissd phenotype in a mutant strain of Escherichia coli B

1968 ◽  
Vol 12 (2) ◽  
pp. 109-116 ◽  
Author(s):  
A. M. Molina ◽  
L. Calegari ◽  
G. Conte
Keyword(s):  
Escherichia Coli ◽  
Cell Membrane ◽  
Mutant Strain ◽  
Minimal Medium ◽  
Specific Requirement ◽  
Resistance Level ◽  
E Coli ◽  
Level Characteristic ◽  
Escherichia Coli B

When an R determinant for streptomycin is transferred into a conditionally streptomycin-dependent E. coli B mutant—which requires in minimal medium either histidine or streptomycin—the latter behaves like a histidineless strain. This phenotype modification shows that the repairing action of streptomycin is prevented. The specific requirement of the strain is not now replaced even by streptomycin concentrations up to 10000 µg/ml at which the conditionally streptomycin-dependent mutant could originally grow, and which are well beyond the resistance level characteristic of the R determinant itself. These data seem to suggest that a reduction in permeability of the cell membrane cannot be held responsible for the phenomenon observed.

scholarly journals Effects of Essential Oils on Escherichia coli Inactivation in Cheese as Described by Meta-Regression Modelling

Foods ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 716
Author(s):  
Beatriz Nunes Silva ◽  
Vasco Cadavez ◽  
José António Teixeira ◽  
Ursula Gonzales-Barron
Keyword(s):  
Escherichia Coli ◽  
Essential Oils ◽  
Meta Analysis ◽  
Inhibitory Effects ◽  
Food Preservatives ◽  
E Coli ◽  
Log Reduction ◽  
Bactericidal Properties ◽  
Study Characteristics ◽  
Meta Regression

The growing intention to replace chemical food preservatives with plant-based antimicrobials that pose lower risks to human health has produced numerous studies describing the bactericidal properties of biopreservatives such as essential oils (EOs) in a variety of products, including cheese. This study aimed to perform a meta-analysis of literature data that could summarize the inactivation of Escherichia coli in cheese achieved by added EOs; and compare its inhibitory effectiveness by application method, antimicrobial concentration, and specific antimicrobials. After a systematic review, 362 observations on log reduction data and study characteristics were extracted from 16 studies. The meta-regression model suggested that pathogenic E. coli is more resistant to EO action than the non-pathogenic type (p < 0.0001), although in both cases the higher the EO dose, the greater the mean log reduction achieved (p < 0.0001). It also showed that, among the factual application methods, EOs’ incorporation in films render a steadier inactivation (p < 0.0001) than when directly applied to milk or smeared on cheese surface. Lemon balm, sage, shallot, and anise EOs showed the best inhibitory outcomes against the pathogen. The model also revealed the inadequacy of inoculating antimicrobials in cheese purposely grated for performing challenge studies, as this non-realistic application overestimates (p < 0.0001) the inhibitory effects of EOs.

scholarly journals Flexible Metabolism and Suppression of Latent Enzymes Are Important forEscherichia coliAdaptation to Diverse Environments within the Host

2019 ◽  
Vol 201 (16) ◽  
Author(s):  
Christopher J. Alteri ◽  
Stephanie D. Himpsl ◽  
Allyson E. Shea ◽  
Harry L. T. Mobley
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Pyruvate Kinase ◽  
Tricarboxylic Acid Cycle ◽  
Financial Burden ◽  
Content Type ◽  
Bacterial Physiology ◽  
E Coli ◽  
Growth Requirements ◽  
Tract Infections

ABSTRACTBacterial metabolism is necessary for adaptation to the host microenvironment. Flexible metabolic pathways allow uropathogenicEscherichia coli(UPEC) to harmlessly reside in the human intestinal tract and cause disease upon extraintestinal colonization.E. coliintestinal colonization requires carbohydrates as a carbon source, while UPEC extraintestinal colonization requires gluconeogenesis and the tricarboxylic acid cycle. UPEC containing disruptions in two irreversible glycolytic steps involving 6-carbon (6-phosphofructokinase;pfkA) and 3-carbon (pyruvate kinase;pykA) substrates have no fitness defect during urinary tract infection (UTI); however, both reactions are catalyzed by isozymes: 6-phosphofructokinases Pfk1 and Pfk2, encoded bypfkAandpfkB, and pyruvate kinases Pyk II and Pyk I, encoded bypykAandpykF. UPEC strains lacking one or both phosphofructokinase-encoding genes (pfkBandpfkA pfkB) and strains lacking one or both pyruvate kinase genes (pykFandpykA pykF) were investigated to determine their regulatory roles in carbon flow during glycolysis by examining their fitness during UTI andin vitrogrowth requirements. Loss of a single phosphofructokinase-encoding gene has no effect on fitness, while thepfkA pfkBdouble mutant outcompeted the parental strain in the bladder. A defect in bladder and kidney colonization was observed with loss ofpykF, while loss ofpykAresulted in a fitness advantage. ThepykA pykFmutant was indistinguishable from wild-typein vivo, suggesting that the presence of Pyk II rather than the loss of Pyk I itself is responsible for the fitness defect in thepykFmutant. These findings suggest thatE. colisuppresses latent enzymes to survive in the host urinary tract.IMPORTANCEUrinary tract infections are the most frequently diagnosed urologic disease, with uropathogenicEscherichia coli(UPEC) infections placing a significant financial burden on the health care system by generating more than two billion dollars in annual costs. This, in combination with steadily increasing antibiotic resistances to present day treatments, necessitates the discovery of new antimicrobial agents to combat these infections. By broadening our scope beyond the study of virulence properties and investigating bacterial physiology and metabolism, we gain a better understanding of how pathogens use nutrients and compete within host microenvironments, enabling us to cultivate new therapeutics to exploit and target pathogen growth requirements in a specific host environment.

scholarly journals DIFFERENTIAL INHIBITORY EFFECTS OF CHOLERA TOXOIDS AND GANGLIOSIDE ON THE ENTEROTOXINS OF VIBRIO CHOLERAE AND ESCHERICHIA COLI

1973 ◽  
Vol 137 (4) ◽  
pp. 1009-1023 ◽  
Author(s):  
Nathaniel F. Pierce
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Competitive Inhibitor ◽  
Inhibitory Effect ◽  
Inhibitory Effects ◽  
E Coli ◽  
Stable Component ◽  
Heat Stable ◽  
Cholera Enterotoxin ◽  
Gut Mucosa

Natural cholera toxoid appears to act as a competitive inhibitor of cholera enterotoxin and is thus a useful tool for studying the interaction of cholera enterotoxin with cell membranes. Cholera enterotoxin binds to gut mucosa more rapidly than does its natural toxoid. Once binding occurs, however, it appears to be prolonged for both materials. Formalinized cholera toxoid has no inhibitory effect upon cholera enterotoxin. Enterotoxic activity, ability to bind to gut mucosa, and antitoxigenicity appear to be independent properties of cholera enterotoxin. Natural cholera toxoid does not inhibit Escherichia coli enterotoxin, indicating that although the two enterotoxins activate the same mucosal secretory mechanism they occupy different binding sites in the mucosa. Ganglioside, which may be the mucosal receptor of cholera enterotoxin, is highly efficient in deactivating cholera enterotoxin. By contrast, ganglioside is relatively inefficient in deactivating heat-labile E. coli enterotoxin and is without effect upon the heat-stable component of E. coli enterotoxin. These findings suggest that ganglioside is not likely to be the mucosal receptor for E. coli enterotoxin. Differences in cellular binding of E. coli and cholera enterotoxins may explain, at least in part, the marked differences in the time of onset and duration of their effects upon gut secretion.

Note: Effect of Some Spice Extracts on Bacterial Inhibition

2003 ◽  
Vol 9 (5) ◽  
pp. 353-358 ◽  
Author(s):  
O. Sagdic ◽  
A. G. Karahan ◽  
M. Ozcan ◽  
G. Ozkan
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Yersinia Enterocolitica ◽  
Inhibition Zone ◽  
Bacterial Strains ◽  
Inhibitory Effects ◽  
Antibacterial Effects ◽  
E Coli ◽  
Inhibition Zones ◽  
Mrs Broth

Eighteen extracts of spices commonly consumed worldwide and grown naturally in Turkey were tested against twenty three bacterial strains to compare their antibacterial effects with eleven antibiotics. Eight pathogens and fifteen lactobacilli isolated from chick intestine were used as the test microorganisms. Pathogens (six different Staphylococcus aureus strains, Escherichia coli ATCC 25922 and Yersinia enterocolitica ATCC 1501) were grown in Nutrient broth and lactobacilli in MRS broth. Hop extracts formed inhibition zones against S. aureus strains of upto 36 mm. Inhibitory effects of hop extracts against S. aureuswere generally higher than that of erythromycin as antibiotic. Helichrysum compactum extract produced an inhibition zone of 23mm to E. coli ATCC 25922 and 26mm to Y. enterocolitica ATCC 1501. Helichrysum compactum extract inhibited the growth of Y. enterocolitica ATCC 1501 more than other spice extracts. While inhibition zones of these extracts against lactobacilli were found smaller than on S. aureus strains, inhibition zones of the same extracts against lactobacilli were found similar to those of E. coli ATCC 25922 and Y. enterocolitica ATCC 1501.

Phosphorylation is switch of L-type pyruvate kinase allostery

2010 ◽  
Vol 5 (2) ◽  
pp. 135-142 ◽  
Author(s):  
Ilona Faustova ◽  
Aleksei Kuznetsov ◽  
Erkki Juronen ◽  
Mart Loog ◽  
Jaak Järv
Keyword(s):  
Pyruvate Kinase ◽  
Hill Coefficient ◽  
Stoichiometric Ratio ◽  
Allosteric Enzyme ◽  
E Coli ◽  
Glycolysis Regulation ◽  
Regulatory Phosphorylation ◽  
Pyruvate Kinase Isoenzymes ◽  
Dependent Protein ◽  
The Hill

AbstractAmong four pyruvate kinase isoenzymes, M1, M2, R and L, only M1 is considered as a nonallosteric enzyme. However, here we show that the non-phosphorylated L-type pyruvate kinase (L-PK) is also a non-allosteric enzyme with respect to its substrate phosphoenolpyruvate (PEP). The allosteric catalytic properties of L-PK are switched on through phosphorylation by cAMP-dependent protein kinase. The non-phosphorylated enzyme was produced by expressing the rat L-PK in E. coli, as the bacterium does not have mammalian-type protein kinases. The resulting tetrameric protein was phosphorylated with a stoichiometric ratio of one mole of phosphate per one L-PK monomer. Activity of the phosphorylated enzyme was allosterically regulated by PEP with the Hill coefficient n=2.5. It was observed that allostery was engaged by phosphorylation of the first subunit in the tetrameric enzyme, while further phosphorylation only modulated this effect. The discovered switching between non-allosteric and allosteric forms of L-PK and the possibility of modulating the allostery by phosphorylation are important for understanding of the interrelationship between allostery and the regulatory phosphorylation in general, and may have implication for further analysis of glycolysis regulation in the liver.

Enhanced Inhibition of Escherichia coli O157:H7 by Lysozyme and Chelators

2003 ◽  
Vol 66 (10) ◽  
pp. 1783-1789 ◽  
Author(s):  
J. S. BOLAND ◽  
P. M. DAVIDSON ◽  
J. WEISS
Keyword(s):  
Escherichia Coli ◽  
Inhibitory Activity ◽  
Chelating Agents ◽  
Inhibitory Effect ◽  
Escherichia Coli O157 ◽  
Inhibitory Effects ◽  
Antimicrobial Spectrum ◽  
E Coli ◽  
Dilution Assay ◽  
Microbroth Dilution

This study examined the effects of three chelating agents (EDTA, disodium pyrophosphate [DSPP], and pentasodium tripolyphosphate [PSTPP]) on the inhibition of the growth of Escherichia coli O157:H7 by lysozyme. The objective of this study was to identify replacement chelators that exhibit synergistic properties similar to those of EDTA. The inhibitory effects of EDTA at 300 to 1,500 μg/ml and of DSPP and PSTPP at 3,000 to 15,000 μg/ml in combination with lysozyme at 200 to 600 μg/ml for up to 48 h at pHs of 6.0, 7.0, and 8.0 on four strains of E. coli O157:H7 was studied with the use of a microbroth dilution assay. The addition of EDTA enhanced lysozyme's inhibitory effect on strains of E. coli O157:H7. EDTA at ≥300 μg/ml combined with lysozyme at 200 to 600 μg/ml was sufficient to inhibit the growth of the strains at pHs of 6.0 and 8.0. At pH 7.0, lysozyme at 200 to 600 μg/ml and EDTA concentrations of ≥1,000 μg/ml were effective in inhibiting three of the four strains. DSPP at pH 6.0 was inhibitory at ≥10,000 μg/ml when combined with lysozyme at 200 to 300 μg/ml. In contrast, PSTPP increased the inhibitory activity of lysozyme more effectively at pH 8.0. Lysozyme at 200 to 600 μg/ml was effective against two strains of E. coli O157:H7 when used in conjunction with PSTPP at ≥5,000 μg/ml. The remaining strains were inhibited by PSTPP at ≥10,000 μg/ml. Our results indicate that inhibition occurred with each lysozyme-chelator combination, but the concentrations of phosphates required to increase the antimicrobial spectrum of lysozyme against E. coli O157:H7 were higher than the EDTA concentrations required to achieve the same effect.

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