Some biochemical effects of 4-deoxy-4-fluoro-D-glucose on Escherichia coli

1977 ◽  
Vol 55 (8) ◽  
pp. 911-915 ◽  
Author(s):  
N. F. Taylor ◽  
Li-Yu Louie
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Catabolite Repression ◽  
Dry Weight ◽  
Resting Cells ◽  
Phosphotransferase System ◽  
E Coli ◽  
Biochemical Effects ◽  
Inhibition Of Growth ◽  
Uncompetitive Inhibitor

The uptake of 4-deoxy-4-fluoro-D-glucose (4FG), without subsequent catabolism, by resting cells of Escherichia coli (ATCC 11775) is 0.06 mg/mg dry weight. In frozen–thawed cells of this organism, 4FG is a substrate for the phosphoenolpyruvate phosphotransferase system with a rate of phosphorylation twice that found for the isomeric 3-deoxy-3-fluoro-D-glucose. 4FG is not a carbon source for growth of this organism and it inhibits the extent of growth of cells in the presence of glucose. The inhibition of growth of E. coli K12 on lactose by 4FG is also observed and this is considered to be consistent with the fact that 4FG is an uncompetitive inhibitor of β-galactosidase (EC 3.2.1.23) activity and that 4FG or 4-deoxy-4-fluoro-D-glucose-6-phosphate repress β-galactosidase synthesis. These results support the view that catabolite repression may be produced by compounds which are not necessarily metabolised further than hexose-6-phosphates.

scholarly journals Effect of deregulation of repressor-specific carbon catabolite repression on carbon source consumption in Escherichia coli

2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Hyeon Jeong Seong ◽  
Yu-Sin Jang
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Catabolite Repression ◽  
Sole Carbon Source ◽  
Carbon Catabolite Repression ◽  
Cell Factory ◽  
Essential Information ◽  
E Coli ◽  
Marine Biomass ◽  
Galactose Utilization

AbstractEscherichia coli has been used as a host to construct the cell factory for biobased production of chemicals from renewable feedstocks. Because galactose is found in marine biomass as a major component, the strategy for galactose utilization in E. coli has been gained more attention. Although galactose and glucose co-fermentation has been reported using the engineered E. coli strain, few reports have covered fermentation supplemented with galactose as a sole carbon source in the mutant lacking the repressor-specific carbon catabolite repression (CCR). Here, we report the effects of the deregulation of the repressor-specific CCR (galR− and galS−) in fermentation supplemented with galactose as a sole carbon source, using the engineered E. coli strains. In the fermentation using the galR− and galS− double mutant (GR2 strain), an increase of rates in sugar consumption and cell growth was observed compared to the parent strain. In the glucose fermentation, wild-type W3110 and its mutant GR2 and GR2PZ (galR−, galS−, pfkA−, and zwf−) consumed sugar at a higher rate than those values obtained from galactose fermentation. However, the GR2P strain (galR−, galS−, and pfkA−) showed no difference between fermentations using glucose and galactose as a sole carbon source. This study provides essential information for galactose fermentation using the CCR-deregulated E. coli strains.

scholarly journals Studies on the lipid content and phosphate requirement of glucose- and acetate-grown Escherichia coli

1968 ◽  
Vol 110 (4) ◽  
pp. 775-781 ◽  
Author(s):  
A. P. Damoglou ◽  
E A Dawes
Keyword(s):  
Escherichia Coli ◽  
Energy Source ◽  
Carbon Source ◽  
Lipid Content ◽  
Dry Weight ◽  
The Other ◽  
Ammonium Salts ◽  
Phosphate Limitation ◽  
E Coli ◽  
High Phosphate

1. The phosphate requirement, i.e. the concentration of inorganic orthophosphate that just ceases to be limiting for growth, of Escherichia coli N.C.T.C. 5928 was determined for growth in ammonium–salts media containing glucose or acetate as the carbon and energy source, and compared with that of six other strains of E. coli. 2. The phosphate requirement for E. coli N.C.T.C. 5928 growing on acetate was about ten times that for growth on glucose, but this difference was not observed with any of the other strains. 3. After about 40 generations' growth on acetate with phosphate limitation in a chemostat, the phosphate requirement of the cells gradually decreased until it was equivalent to that of the glucose-grown organism; a single passage through glucose batch culture sufficed to restore the original high phosphate requirement, indicating a permeability phenomenon. 4. The lipid content of E. coli N.C.T.C. 5928 grown on glucose or acetate was measured isotopically by fractionation of cells grown on inorganic [32P]orthophosphate and gravimetrically after extraction from the cells by three different methods; change of carbon source from glucose to acetate did not affect the lipid content, which remained constant at 8–9% of the bacterial dry weight.

Biofilm Growth of Escherichia coli Is Subject to cAMP-Dependent and cAMP-Independent Inhibition

2015 ◽  
Vol 25 (2-3) ◽  
pp. 209-225 ◽  
Author(s):  
Sarah L. Sutrina ◽  
Kia Daniel ◽  
Michael Lewis ◽  
Naomi T. Charles ◽  
Cherysa K.E. Anselm ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Catabolite Repression ◽  
Ph Effects ◽  
Wild Type ◽  
Inhibitory Effects ◽  
Phosphotransferase System ◽  
Biofilm Growth ◽  
E Coli ◽  
Low Concentrations ◽  
High Concentrations

We established that <i>Escherichia coli </i>strain 15 (ATCC 9723) produces both curli and cellulose, and forms robust biofilms. Since this strain is wild type with respect to the phosphoenolpyruvate:sugar phosphotransferase system (PTS), it is an ideal strain in which to investigate the effects of the PTS on the biofilm growth of <i>E. coli</i>. We began by looking into the effects of PTS and non-PTS sugars on the biofilm growth of this strain. All the sugars tested tended to activate biofilm growth at low concentrations but to inhibit biofilm growth at high concentrations. Acidification of the medium was an inhibitory factor in the absence of buffer, but buffering to prevent a pH drop did not prevent the inhibitory effects of the sugars. The concentration at which inhibition set in varied from sugar to sugar. For most sugars, cyclic (c)AMP counteracted the inhibition at the lowest inhibitory concentrations but became ineffective at higher concentrations. Our results suggest that cAMP-dependent catabolite repression, which is mediated by the PTS in <i>E. coli</i>, plays a role in the regulation of biofilm growth in response to sugars. cAMP-independent processes, possibly including Cra, also appear to be involved, in addition to pH effects.

Isolation and characterization of catabolite repression-resistant mutants of Escherichia coli

1977 ◽  
Vol 23 (10) ◽  
pp. 1384-1393 ◽  
Author(s):  
Glen D. Armstrong ◽  
Hiroshi Yamazaki
Keyword(s):  
Escherichia Coli ◽  
Catabolite Repression ◽  
Coli Strain ◽  
Wild Type ◽  
Phosphotransferase System ◽  
E Coli ◽  
Isolation And Characterization ◽  
In The Wild ◽  
Resistant Mutants

A method has been developed for the isolation of Escherichia coli mutants which are resistant to catabolite repression. The method is based on the fact that a mixture of glucose and gluconate inhibits the development of chemotactic motility in the wild type, but not in the mutants. A motile E. coli strain was mutagenized and grown in glucose and gluconate. Mutants which were able to swim into a tube containing a chemotactic attractant (aspartic acid) were isolated. Most of these mutants were able to produce β-galactosidase in the presence of glucose and gluconate and were normal in their ability to degrade adenosine 3′,5′-cyclic monophosphate. Some of these mutants were defective in the glucose phosphotransferase system.

Inactivation of the PTS as a Strategy to Engineer the Production of Aromatic Metabolites in Escherichia coli

2015 ◽  
Vol 25 (2-3) ◽  
pp. 195-208 ◽  
Author(s):  
Susy Beatriz Carmona ◽  
Fabián Moreno ◽  
Francisco Bolívar ◽  
Guillermo Gosset ◽  
Adelfo Escalante
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Transport Systems ◽  
Phosphotransferase System ◽  
Adaptive Laboratory Evolution ◽  
E Coli ◽  
Inducer Exclusion ◽  
Genetic Modifications ◽  
Phosphate Donor ◽  
Selection Of

Laboratory and industrial cultures of <i>Escherichia coli</i> employ media containing glucose which is mainly transported and phosphorylated by the phosphotransferase system (PTS). In these strains, 50% of the phosphoenolpyruvate (PEP), which results from the catabolism of transported glucose, is used as a phosphate donor for its phosphorylation and translocation by the PTS. This characteristic of the PTS limits the production of industrial biocommodities that have PEP as a precursor. Furthermore, when <i>E. coli</i> is exposed to carbohydrate mixtures, the PTS prevents expression of catabolic and non-PTS transport genes by carbon catabolite repression and inducer exclusion. In this contribution, we discuss the main strategies developed to overcome these potentially limiting effects in production strains. These strategies include adaptive laboratory evolution selection of PTS<sup>-</sup> Glc<sup>+</sup> mutants, followed by the generation of strains that recover their ability to grow with glucose as a carbon source while allowing the simultaneous consumption of more than one carbon source. We discuss the benefits of using alternative glucose transport systems and describe the application of these strategies to <i>E. coli</i> strains with specific genetic modifications in target pathways. These efforts have resulted in significant improvements in the production of diverse biocommodities, including aromatic metabolites, biofuels and organic acids.

scholarly journals Rsd balances (p)ppGpp level by stimulating the hydrolase activity of SpoT during carbon source downshift inEscherichia coli

2018 ◽  
Vol 115 (29) ◽  
pp. E6845-E6854 ◽  
Author(s):  
Jae-Woo Lee ◽  
Young-Ha Park ◽  
Yeong-Jae Seok
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Control Mechanism ◽  
Stringent Response ◽  
Hydrolase Activity ◽  
Inescherichia Coli ◽  
Phosphotransferase System ◽  
E Coli ◽  
Cellular Concentration ◽  
Spot Activity

Bacteria respond to nutritional stresses by changing the cellular concentration of the alarmone (p)ppGpp. This control mechanism, called the stringent response, depends on two enzymes, the (p)ppGpp synthetase RelA and the bifunctional (p)ppGpp synthetase/hydrolase SpoT inEscherichia coliand related bacteria. Because SpoT is the only enzyme responsible for (p)ppGpp hydrolysis in these bacteria, SpoT activity needs to be tightly regulated to prevent the uncontrolled accumulation of (p)ppGpp, which is lethal. To date, however, no such regulation of SpoT (p)ppGpp hydrolase activity has been documented inE. coli. In this study, we show that Rsd directly interacts with SpoT and stimulates its (p)ppGpp hydrolase activity. Dephosphorylated HPr, but not phosphorylated HPr, of the phosphoenolpyruvate-dependent sugar phosphotransferase system could antagonize the stimulatory effect of Rsd on SpoT (p)ppGpp hydrolase activity. Thus, we suggest that Rsd is a carbon source-dependent regulator of the stringent response inE. coli.

scholarly journals Relaxation of catabolite repression in streptomycin-dependent Escherichia coli

1969 ◽  
Vol 111 (3) ◽  
pp. 279-286 ◽  
Author(s):  
M. B. Coukell ◽  
W. J. Polglase
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Catabolite Repression ◽  
Citrate Synthase ◽  
Wild Type ◽  
Experimental Conditions ◽  
E Coli ◽  
Equivalent Quantity ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Escherichia Coli B

Acetohydroxy acid synthetase, which is sensitive to catabolite repression in wild-type Escherichia coli B, was relatively resistant to this control in a streptomycin-dependent mutant. The streptomycin-dependent mutant was found to be inducible for β-galactosidase in the presence of glucose, although repression of β-galactosidase by glucose occurred under experimental conditions where growth of the streptomycin-dependent mutant was limited. Additional glucose-sensitive enzymes of wild-type E. coli B (citrate synthase, fumarase, aconitase and isocitrate dehydrogenase) were found to be insensitive to the carbon source in streptomycin-dependent mutants: these enzymes were formed by streptomycin-dependent E. coli B in equivalent quantities when either glucose or glycerol was the carbon source. Two enzymes, glucokinase and glucose 6-phosphate dehydrogenase, that are glucose-insensitive in wild-type E. coli B were formed in equivalent quantity on glucose or glycerol in both streptomycin-sensitive and streptomycin-dependent E. coli B. The results indicate a general decrease or relaxation of catabolite repression in the streptomycin-dependent mutant. The yield of streptomycin-dependent cells from glucose was one-third less than that of the streptomycin-sensitive strain. We conclude that the decreased efficiency of glucose utilization in streptomycin-dependent E. coli B is responsible for the relaxation of catabolite repression in this mutant.

scholarly journals Inducible phosphoenolpyruvate-dependent hexose phosphotransferase activities in Escherichia coli

1972 ◽  
Vol 128 (5) ◽  
pp. 1339-1344 ◽  
Author(s):  
H. L. Kornberg ◽  
Richard E. Reeves
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Phosphotransferase System ◽  
E Coli ◽  
Low Glucose ◽  
Specific Activities

1. A method is described for measuring the rate of phosphoenolpyruvate-dependent phosphotransferase activity for a variety of hexoses in toluene-treated suspensions of Escherichia coli. 2. The specific activities of the phosphotransferases that catalyse the phosphorylation of hexoses are greatly affected by the carbon source for growth. 3. In all strains of E. coli tested, fructose phosphotransferase activity is induced by growth on fructose. 4. Strains of E. coli differ greatly in the rate at which they phosphorylate glucose, but all strains possess at least a low glucose phosphotransferase activity under any tested condition of growth. Glucose phosphotransferase activity is further induced by growth on glucose; this does not occur in a mutant that lacks the ability to take up methyl α-d-[14C]glucopyranoside and hence grows poorly on glucose. 5. When growing on fructose, two strains of E. coli synthesize the inducible glucose phosphotransferase system gratuitously, and to specific activities higher than observed during growth on glucose. A phosphotransferase catalysing the phosphorylation of mannose is similarly induced.

Inhibition of Escherichia coli Trimethylamine-N-oxide Reductase by Food Preservatives1

1982 ◽  
Vol 45 (3) ◽  
pp. 241-243 ◽  
Author(s):  
M. KRUK ◽  
J. S. LEE
Keyword(s):  
Escherichia Coli ◽  
Benzoic Acid ◽  
Sorbic Acid ◽  
Reductase Activity ◽  
Resting Cells ◽  
E Coli

Trimethylamine-N-oxide (TMA-O) reductase activity of resting cells of Escherichia coli was inhibited by tetrasodium ethylenediaminetetraacetate (Na4EDTA), benzoic acid (BA and methylparaben (MP). The 50% inhibitory concentrations of Na4EDTA, BA and MP were 20.2, 1.2 and 32.4 mM, respectively. BA at pH 6.5 or below most effectively inhibited the TMA-O reductase. Sorbic acid (SA), up to 0.70 mM, had no effect on TMA-O reductase activity, but SA inhibited the growth and subsequent TMA production in E. coli at or above 0.3S mM.

Mannose utilization in Escherichia coli requires cyclic AMP but not an exogenous inducer

1980 ◽  
Vol 26 (12) ◽  
pp. 1508-1511 ◽  
Author(s):  
Ann D. E. Fraser ◽  
Hiroshi Yamazaki
Keyword(s):  
Escherichia Coli ◽  
Carbon Source ◽  
Cyclic Amp ◽  
Adenylyl Cyclase ◽  
Sole Carbon Source ◽  
Receptor Protein ◽  
Deficient Mutant ◽  
Phosphotransferase System ◽  
Cyclic Amp Receptor Protein ◽  
Cyclic Monophosphate

It has not been clarified whether the utilization of mannose by Escherichia coli requires adenosine 3′,5′-cyclic monophosphate (cyclic AMP). Using an adenylyl cyclase deficient mutant (CA8306B) and a cyclic AMP receptor protein (CRP) deficient mutant (5333B) we have shown that the utilization of mannose is dependent on the cyclic AMP–CRP complex. 2-Deoxyglucose (DG) is a nonmetabolizable glucose analog specific for the phosphotransferase system (PTS) which transports mannose (termed here PTSM). Growth of CA8306B on glycerol is unaffected by addition of the analog, whereas growth of the strain on glycerol plus cyclic AMP ceases im mediately upon addition of DG. These results suggest that the formation of PTSM is dependent on cyclic AMP. In addition, CA8306B grown on glycerol plus cyclic AMP can immediately utilize mannose when transferred to a medium containing mannose as a sole carbon source, whereas the same strain grown on glycerol without cyclic AMP cannot utilize mannose when so transferred. These results suggest that the formation of PTSM does not require an exogenous inducer.

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