DNA-Binding Protein Dps Protects <i>Escherichia coli</i> Cells against Multiple Stresses during Desiccation

Gradual dehydration is one of the frequent lethal yet poorly understood stresses that bacterial cells constantly face in the environment when their micro ecotopes dry out, as well as in industrial processes. Bacteria successfully survive extreme desiccation through complex rearrangements at the structural, physiological, and molecular levels, in which proteins are involved. The DNA-binding protein Dps has previously been shown to protect bacterial cells from many adverse effects. In our work, using engineered genetic models of E. coli to produce bacterial cells with overproduction of Dps protein, the protective function of Dps protein under multiple desiccation stresses was demonstrated for the first time. It was shown that the titer of viable cells after rehydration in the experimental variants with Dps protein overexpression was 1.5–8.5 times higher. Scanning electron microscopy was used to show a change in cell morphology upon rehydration. It was also proved that immobilization in the extracellular matrix, which is greater when the Dps protein is overexpressed, helps the cells survive. Transmission electron microscopy revealed disruption of the crystal structure of DNA–Dps crystals in E. coli cells that underwent desiccation stress and subsequent watering. Coarse-grained molecular dynamics simulations showed the protective function of Dps in DNA–Dps co-crystals during desiccation. The data obtained are important for improving biotechnological processes in which bacterial cells undergo desiccation

Biocides with Controlled Degradation for Environmentally Friendly and Cost-Effective Fecal Sludge Management

Didecyldimethylammonium chloride (DDAC) and polyhexamethylene guanidine (PHMG) exhibit high antimicrobial activity and are widely used as biocidal agents in chemical toilet additives for the management of fecal sludge (FS). Disposal of such biocide-treated FS to a wastewater treatment plant (WWTP) is a major environmental problem. It is possible to reduce environmental damage through the use of biocidal agents, which easily decompose after performing their main biocidal functions. In this work, it is proposed to use the fact of a gradual increase in pH of FS from the initial 7.5 to 9.0–10.0 due to the decomposition of urea. Six biocidal compounds were selected that are capable of rapidly degrading in an alkaline environment and one that naturally degrades upon prolonged incubation. Four of them: bronopol (30 mg/L), DBNPA (500 mg/L), Sharomix (500 mg/L), and sodium percarbonate (6000 mg/L) have shown promise for environmentally friendly management of FS. In selected dosage, they successfully reduced microbial activity under both aerobic and anaerobic conditions and are cost-effective. After 10 days of incubation, degradation of the biocide occurred as measured by biological oxygen demand (BOD5) in biocide-treated FS. Such FS can be discharged to WWTP without severe damage to the activated sludge process, the need for dilution and additional procedures to neutralize toxicity

Morphological peculiarities of the DNA-protein complexes in starved Escherichia coli cells.

One of the adaptive strategies for the constantly changing conditions of the environment utilized in bacterial cells involves the condensation of DNA in complex with the DNA-binding protein, Dps. With the use of electron microscopy and electron tomography, we observed several morphologically different types of DNA condensation in dormant Escherichia coli cells, namely: nanocrystalline, liquid crystalline, and the folded nucleosome-like. We confirmed the presence of both Dps and DNA in all of the ordered structures using EDX analysis. The comparison of EDX spectra obtained for the three different ordered structures revealed that in nanocrystalline formation the majority of the Dps protein is tightly bound to nucleoid DNA. The dps-null cells contained only one type of condensed DNA structure, liquid crystalline, thus, differing from those with Dps. The results obtained here shed some light on the phenomenon of DNA condensation in dormant prokaryotic cells and on the general problem of developing a response to stress. We demonstrated that the population of dormant cells is structurally heterogeneous, allowing them to respond flexibly to environmental changes. It increases the ability of the whole bacterial population to survive under extreme stress conditions