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Cross-Talk between the Canonical and the Nitrogen-Related Phosphotransferase Systems Modulates Synthesis of the KdpFABC Potassium Transporter in Escherichia coli
Many Proteobacteria possess the regulatory nitrogen-related phosphotransferase system (PTSNtr), which operates in parallel to the transport PTS. PTSNtr is composed of the proteins EINtr and NPr and the final phosphate acceptor EIIANtr. Both PTSs can exchange phosphoryl groups among each other. Proteins governing K+ uptake represent a major target of PTSNtr in Escherichia coli. Nonphosphorylated EIIANtr binds and stimulates the K+ sensor KdpD, which activates expression of the kdpFABC operon encoding a K+ transporter. Here we show that this regulation also operates in an ilvG+ strain ruling out previous concern about interf...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

Regulation of the Utilization of Amino Sugars by Escherichia coli and Bacillus subtilis: Same Genes, Different Control
Amino sugars are dual-purpose compounds in bacteria: they are essential components of the outer wall peptidoglycan (PG) and the outer membrane of Gram-negative bacteria and, in addition, when supplied exogenously their catabolism contributes valuable supplies of energy, carbon and nitrogen to the cell. The enzymes for both the synthesis and degradation of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) are highly conserved but during evolution have become subject to different regulatory regimes. Escherichia coli grows more rapidly using GlcNAc as a carbon source than with GlcN. On the other hand, Bacillus subtilis, but...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

The Phosphotransferase System in Solventogenic Clostridia
The acetone-butanol-ethanol fermentation employing solventogenic clostridia was a major industrial process during the 20th century, but declined for economic reasons. In recent times, interest in the process has been revived due to the perceived potential of butanol as a superior biofuel. Redevelopment of an efficient fermentation process will require a detailed understanding of the physiology of carbohydrate utilization by the bacteria. Genome sequences have revealed that, as in other anaerobes, the phosphotransferase system (PTS) and associated regulatory functions are likely to play an important role in sugar uptake and...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

Inactivation of the PTS as a Strategy to Engineer the Production of Aromatic Metabolites in Escherichia coli
Laboratory and industrial cultures of Escherichia coli 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 E. coli is exposed to carbohydrate mixtures, the PTS prevents expression of catabolic and non-PTS transport genes by carbon catab...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

A Search for Ribonucleic Antiterminator Sites in Bacterial Genomes: Not Only Antitermination?
BglG/LicT-like proteins are transcriptional antiterminators that prevent termination of transcription at intrinsic terminators by binding to ribonucleic antiterminator (RAT) sites and stabilizing an RNA conformation which is mutually exclusive with the terminator structure. The known RAT sites, which are located in intergenic regions of sugar utilization operons, show low sequence conservation but significant structural analogy. To assess the prevalence of RATs in bacterial genomes, we employed bioinformatic tools that describe RNA motifs based on both sequence and structural constraints. Using descriptors with different s...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

Fucose-Mediated Transcriptional Activation of the fcs Operon by FcsR in Streptococcus pneumoniae
In this study, we explore the impact of fucose on the transcriptome of S. pneumoniae D39. The expression of various genes and operons, including the fucose uptake PTS and utilization operon (fcs operon) was altered in the presence of fucose. By means of quantitative RT-PCR and β-galactosidase analysis, we demonstrate the role of the transcriptional regulator FcsR, present upstream of the fcs operon, as a transcriptional activator of the fcs operon. We also predict a 19-bp putative FcsR regulatory site (5′-ATTTGAACATTATTCAAGT-3′) in the promoter region of the fcs operon. The functionality of this predicted FcsR regulat...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

Synthesis and Physicochemical Characterization of D-Tagatose-1-Phosphate: The Substrate of the Tagatose-1-Phosphate Kinase in the Phosphotransferase System-Mediated D-Tagatose Catabolic Pathway of Bacillus licheniformis
We report the first enzymatic synthesis of D-tagatose-1-phosphate (Tag-1P) by the multicomponent phosphoenolpyruvate:sugar phosphotransferase system (PEP-PTS) present in tagatose-grown cells of Klebsiella pneumoniae. Physicochemical characterization by 31P and 1H nuclear magnetic resonance spectroscopy reveals that, in solution, this derivative is primarily in the pyranose form. Tag-1P was used to characterize the putative tagatose-1-phosphate kinase (TagK) of the Bacillus licheniformis PTS-mediated D-tagatose catabolic pathway (Bli-TagP). For this purpose, a soluble protein fusion was obtained with the 6 His-tagged trigge...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

PTS 50: Past, Present and Future, or Diauxie Revisited
Past: The title ‘PTS 50 or The PTS after 50 years' relies on the first description in 1964 of the phosphoenolpyruvate-dependent carbohydrate:phosphotransferase system (PTS) by Kundig, Gosh and Roseman [Proc Natl Acad Sci USA 1964;52:1067-1074]. The system comprised proteins named Enzyme I, HPr and Enzymes II, as part of a novel PTS for carbohydrates in Gram-negative and Gram-positive bacteria, whose ‘biological significance remained unclear'. In contrast, studies which would eventually lead to the discovery of the central role of the PTS in bacterial metabolism had been published since before 1942. They are primarily l...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

The Bacterial Phosphotransferase System: New Frontiers 50 Years after Its Discovery
In 1964, Kundig, Ghosh and Roseman reported the discovery of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), which they subsequently proposed might catalyze sugar transport as well as sugar phosphorylation. What we have learned in the 50 years since its discovery is that, in addition to these primary functions, the PTS serves as a complex protein kinase system that regulates a wide variety of transport, metabolic and mutagenic processes as well as the expression of numerous genes. Recent operon- and genome-sequencing projects have revealed novel PTS protein-encoding genes, many of which have yet to be functi...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

PTS-Mediated Regulation of the Transcription Activator MtlR from Different Species: Surprising Differences despite Strong Sequence Conservation
The hexitol D-mannitol is transported by many bacteria via a phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS). In most Firmicutes, the transcription activator MtlR controls the expression of the genes encoding the D-mannitol-specific PTS components and D-mannitol-1-P dehydrogenase. MtlR contains an N-terminal helix-turn-helix motif followed by an Mga-like domain, two PTS regulation domains (PRDs), an EIIBGat- and an EIIAMtl-like domain. The four regulatory domains are the target of phosphorylation by PTS components. Despite strong sequence conservation, the mechanisms controlling the activity of MtlR ...
Source: Journal of Molecular Microbiology and Biotechnology - July 11, 2015 Category: Microbiology Source Type: research

Caldariomyces fumago DSM1256 Contains Two Chloroperoxidase Genes, Both Encoding Secreted and Active Enzymes
Inspection of transcriptome data from the chloroperoxidase (CPO)-producing fungus Caldariomyces fumago DSM1256 led to the discovery of two distinct CPO mRNA sequences. This strain could be shown to contain the newly identified isogene as well as produce and secrete both isoenzymes. The CPO2 enzyme bears high sequence similarity to the well-characterized CPO (87% identity for the mature proteins). It shows two insertions in the signal peptide and in the C-terminal propeptide, and one deletion in the mature polypeptide close to the C-terminus. Furthermore, it lacks one of the serine residues known to be O-glycosylated in the...
Source: Journal of Molecular Microbiology and Biotechnology - June 30, 2015 Category: Microbiology Source Type: research

In vitro Antifungal Activity and Mechanism of Action of Tea Polyphenols and Tea Saponin against Rhizopus stolonifer
The in vitro antifungal activities and mechanism of action of tea polyphenols (TP), tea saponin (TS) and their combination were evaluated against Rhizopus stolonifer. The results showed that both TP and TS inhibited the mycelial growth in a dose-dependent manner, and their combination at the ratio of 7:3 exhibited synergistic antifungal interaction. We also observed that the treatment of TP or TS significantly induced the production of H2O2 and resulted in membrane lipid peroxidation, thus leading to an increase in cell membrane permeability and the leakage of K+, soluble protein and soluble sugar. Moreover, combining them...
Source: Journal of Molecular Microbiology and Biotechnology - June 30, 2015 Category: Microbiology Source Type: research

Developmental Dynamic Analysis of the Excreted Microbiome of Chickens Using Next-Generation Sequencing
Poultry contamination can be largely attributed to the presence of chicken feces during the production process. Fecal contamination is often found in raw chicken products sold for human consumption. Quantitative analysis of the fecal microbial community of chickens using next-generation sequencing techniques is the focus of this study. Fecal samples were collected from 30 broiler chickens at two time points: days 1 and 35 of development. 454 pyrosequencing was conducted on 16S rRNA extracted from each sample, and microbial population dynamics were investigated using various automated bioinformatics pipelines. Diversity of ...
Source: Journal of Molecular Microbiology and Biotechnology - June 30, 2015 Category: Microbiology Source Type: research

Optimization of Culture Medium for Maximal Production of Spinosad Using an Artificial Neural Network - Genetic Algorithm Modeling
Conclusion: The hybrid ANN/GA approach provides a viable alternative to the conventional RSM approach for the modeling and optimization of fermentation processes.J Mol Microbiol Biotechnol 2015;25:253-261 (Source: Journal of Molecular Microbiology and Biotechnology)
Source: Journal of Molecular Microbiology and Biotechnology - June 30, 2015 Category: Microbiology Source Type: research

Gene Expression of Lytic Endopeptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonads
Development of an efficient expression system for (especially secreted) bacterial lytic enzymes is a complicated task due to the specificity of their action. The substrate for such enzymes is peptidoglycan, the main structural component of bacterial cell walls. For this reason, expression of recombinant lytic proteins is often accompanied with lysis of the producing bacterium. This paper presents data on the construction of an inducible system for expression of the lytic peptidases AlpA and AlpB from Lysobacter sp. XL1 in Pseudomonas fluorescens Q2-87, which provides for the successful secretion of these proteins into the ...
Source: Journal of Molecular Microbiology and Biotechnology - June 30, 2015 Category: Microbiology Source Type: research