Analysis of global control of Escherichia coli carbohydrate uptake

<p>Abstract</p> <p>Background</p> <p>Global control influences the regulation of many individual subsystems by superimposed regulator proteins. A prominent example is the control of carbohydrate uptake systems by the transcription factor Crp in <it>Escherichia coli</it>. A detailed understanding of the coordination of the control of individual transporters offers possibilities to explore the potential of microorganisms e.g. in biotechnology.</p> <p>Results</p> <p>An o.d.e. based mathematical model is presented that maps a physiological parameter – the specific growth rate – to the sensor of the signal transduction unit, here a component of the bacterial phosphotransferase system (PTS), namely EIIA^<it>Crr</it>. The model describes the relation between the growth rate and the degree of phosphorylation of EIIA ^{<it>crr </it>}for a number of carbohydrates by a distinctive response curve, that differentiates between PTS transported carbohydrates and non-PTS carbohydrates. With only a small number of kinetic parameters, the model is able to describe a broad range of experimental steady-state and dynamical conditions.</p> <p>Conclusion</p> <p>The steady-state characteristic presented shows a relationship between the growth rate and the output of the sensor system PTS. The glycolytic flux that is measured by this sensor is a good indicator to represent the nutritional status of the cell.</p

Comparative proteomics of uropathogenic Escherichia coli during growth in human urine identify UCA-like (UCL) fimbriae as an adherence factor involved in biofilm formation and binding to uroepithelial cells

Uropathogenic Escherichia coli (UPEC) are the primary cause of urinary tract infection (UTI) in humans. For the successful colonisation of the human urinary tract, UPEC employ a diverse collection of secreted or surface-exposed virulence factors including toxins, iron acquisition systems and adhesins. In this study, a comparative proteomic approach was utilised to define the UPEC pan and core surface proteome following growth in pooled human urine. Identified proteins were investigated for subcellular origin, prevalence and homology to characterised virulence factors. Fourteen core surface proteins were identified, as well as eleven iron uptake receptor proteins and four distinct fimbrial types, including type 1, P, F1C/S and a previously uncharacterised fimbrial type, designated UCA-like (UCL) fimbriae in this study. These pathogenicity island (PAI)-associated fimbriae are related to UCA fimbriae of Proteus mirabilis, associated with UPEC and exclusively found in members of the E. coli B2 and D phylogroup. We further demonstrated that UCL fimbriae promote significant biofilm formation on abiotic surfaces and mediate specific attachment to exfoliated human uroepithelial cells. Combined, this study has defined the surface proteomic profiles and core surface proteome of UPEC during growth in human urine and identified a new type of fimbriae that may contribute to UTI

Minimizing acetate formation in E. coli fermentations

Escherichia coli remains the best established production organisms in industrial biotechnology. However, during aerobic fermentation runs at high growth rates, considerable amounts of acetate are accumulated as by-product. This by-product has negative effects on growth and protein production. Over the last 20 years, substantial research efforts have been spent to reduce acetate accumulation during aerobic growth of E. coli on glucose. From the onset it was clear that this quest should not be a simple nor uncomplicated one. Simple deletion of the acetate pathway, reduced the acetate accumulation, but instead other by-products were formed. This minireview gives a clear outline of these research efforts and the outcome of them, including bioprocess level approaches and genetic approaches. Recently, the latter seems to have some promising results

Microarray-based screening of differentially expressed genes of E. coli O157:H7 Sakai during preharvest survival on butterhead lettuce

Numerous outbreaks of Escherichia coli O157:H7 have been linked to the consumption of leafy vegetables. However, up to the present, little has been known about E. coli O157:H7's adaptive responses to survival on actively growing (and thus responsive) plants. In this study, whole genome transcriptional profiles were generated from E. coli O157:H7 cells (isolate Sakai, stx-) one hour and two days after inoculation on the leaves of growing butterhead lettuce, and compared with an inoculum control. A total of 273 genes of E. coli O157:H7 Sakai (5.04% of the whole genome) were significantly induced or repressed by at least two-fold (p < 0.01) in at least one of the analyzed time points in comparison with the control. Several E. coli O157:H7 genes associated with oxidative stress and antimicrobial resistance were upregulated, including the iron-sulfur cluster and the multiple antibiotic resistance (mar) operon, whereas the Shiga toxin virulence genes were downregulated. Nearly 40% of the genes with significantly different expression were poorly characterized genes or genes with unknown functions. These genes are of special interest for future research as they may play an important role in the pathogens' adaptation to a lifestyle on plants. In conclusion, these findings suggest that the pathogen actively interacts with the plant environment by adapting its metabolism and responding to oxidative stress

Substrate induction and glucose repression of maltose utilization by Streptomyces coelicolor A3(2) is controlled by malR, a member of the lacI-galR family of regulatory genes

malR of Strepomyces coelicolor A3(2) encodes a homologue of the Lacl/Galr family of repressor proteins, and is divergently transcribed from the malEFG gene cluster, which encodes components of an ATP-dependent transport system that is required for maltose utilization. Transcription of malE was induced by maltose and repressed by glucose. Disruption or deletion of malR resulted in constitutive, glucose-insensitive malE transcription at a level markedly above that observed in the parental malR+ strain, and overproduction of MalR prevented growth on maltose as carbon source. Consequently, MalR plays a crucial role in both substrate induction and glucose repression of maltose utilization. MalR is expressed from a single promoter with transcription initiating at the first G of the predicted GTG translataion start codon

Effects of deletion of the Streptococcus pneumoniae lipoprotein diacylglyceryl transferase gene lgt on ABC transporter function and on growth in vivo

Lipoproteins are an important class of surface associated proteins that have diverse roles and frequently are involved in the virulence of bacterial pathogens. As prolipoproteins are attached to the cell membrane by a single enzyme, prolipoprotein diacylglyceryl transferase (Lgt), deletion of the corresponding gene potentially allows the characterisation of the overall importance of lipoproteins for specific bacterial functions. We have used a Δlgt mutant strain of Streptococcus pneumoniae to investigate the effects of loss of lipoprotein attachment on cation acquisition, growth in media containing specific carbon sources, and virulence in different infection models. Immunoblots of triton X-114 extracts, flow cytometry and immuno-fluorescence microscopy confirmed the Δlgt mutant had markedly reduced lipoprotein expression on the cell surface. The Δlgt mutant had reduced growth in cation depleted medium, increased sensitivity to oxidative stress, reduced zinc uptake, and reduced intracellular levels of several cations. Doubling time of the Δlgt mutant was also increased slightly when grown in medium with glucose, raffinose and maltotriose as sole carbon sources. These multiple defects in cation and sugar ABC transporter function for the Δlgt mutant were associated with only slightly delayed growth in complete medium. However the Δlgt mutant had significantly reduced growth in blood or bronchoalveolar lavage fluid and a marked impairment in virulence in mouse models of nasopharyngeal colonisation, sepsis and pneumonia. These data suggest that for S. pneumoniae loss of surface localisation of lipoproteins has widespread effects on ABC transporter functions that collectively prevent the Δlgt mutant from establishing invasive infection

Integrating dark and light biohydrogen production strategies: towards the hydrogen economy

Biological methods of hydrogen production are preferable to chemical methods because of the possibility to use sunlight, CO2 and organic wastes as substrates for environmentally benign conversions, under moderate conditions. By combining different microorganisms with different capabilities, the individual strengths of each may be exploited and their weaknesses overcome. Mechanisms of bio-hydrogen production are described and strategies for their integration are discussed. Dual systems can be\ud divided broadly into wholly light-driven systems (with microalgae/cyanobacteria as the 1st stage) and partially light-driven systems (with a dark, fermentative initial reaction). Review and evaluation of published data suggests that the latter type of system holds greater promise for industrial application. This is because the calculated land area required for a wholly light-driven dual system would be too large for either centralised (macro-) or decentralised(micro-) energy generation. The potential contribution to the hydrogen economy of partially light-driven dual systems is overviewed alongside that of other biofuels such as bio-methane and bio-ethanol

Effect of cra Gene Mutation on the Metabolism of Escherichia Coli for a Mixture of Multiple Carbon Sources

The major player for catabolite repression is the phosphotransferase systems (PTSs) and cAMP-Crp. Moreover, Cra controls the carbon flow in the metabolic network. In the present research, the effect of modulating cra gene (Δcra) on the consumption of multiple carbon sources such as glucose and fructose (as well as xylose) was investigated under both aerobic and anaerobic conditions. It was shown that glucose and fructose could be co-metabolized with fructose consumed faster than glucose in cra mutant under both aerobic and anaerobic conditions. It was also implied that cra mutant consumed higher amount of total carbon sources, which contributed to the highest metabolite production as compared to the wild type strain. Thus, cra mutant can be a good candidate for the efficient utilization of multiple carbon sources such as glucose and fructose, where xylose consumption was repressed by catabolite repression. The overall regulation mechanisms were clarified based on fermentation data and gene transcript analysis