RNA-Binding Protein Hfq: A Role in Cellulose Decomposition

ObjectiveTo study the function of the RNA-binding protein Hfq in Bacillus subtilis cellulose decomposition.ResultsIn the medium with sodium carboxymethylcellulose (Na-CMC) as the sole carbon source, the knockout of Hfq resulted in a 38.0% ± 2.1% and 76.6% ± 7.1% decrease in cellulose hydrolysis ability and cellulase activity, respectively. The results of real-time quantitative PCR revealed that several cellulase genes (eglS, bglA, and bglC) were significantly downregulated in the Hfq knockout strain. The isogenic ΔHfq complemented strain recovered the cellulose hydrolysis ability, cellulase activity, and expression level of cellulase genes. In addition, the survival of Hfq mutant in stationary phase was significantly affected. ConclusionRNA-binding protein Hfq is involved in the regulation of cellulose hydrolysis ability, cellulase activity, cellulase gene expression, and stationary phase survival.

Reprioritization of biofilm metabolism is associated with nutrient adaptation and long-term survival of Haemophilus influenzae

Nontypeable Haemophilus influenzae (NTHI) is a human-restricted pathogen with an essential requirement for heme–iron acquisition. We previously demonstrated that microevolution of NTHI promotes stationary phase survival in response to transient heme–iron restriction. In this study, we examine the metabolic contributions to biofilm formation using this evolved NTHI strain, RM33. Quantitative analyses identified 29 proteins, 55 transcripts, and 31 metabolites that significantly changed within in vitro biofilms formed by RM33. The synthesis of all enzymes within the tryptophan and glycogen pathways was significantly increased in biofilms formed by RM33 compared with the parental strain. In addition, increases were observed in metabolite transport, adhesin production, and DNA metabolism. Furthermore, we observed pyruvate as a pivotal point in the metabolic pathways associated with changes in cAMP phosphodiesterase activity during biofilm formation. Taken together, changes in central metabolism combined with increased stores of nutrients may serve to counterbalance nutrient sequestration.

A Bumpy Pathway to Stationary-Phase Survival in Bacillus subtilis

ABSTRACT Bacillus subtilis cells can mount a number of responses to nutritional deprivation but ultimately either form dormant spores or enter a metabolically quiescent state. In a recent article (mBio 10:e01414-19, https://doi.org/10.1128/mBio.01414-19, 2019), R. Hashuel and S. Ben-Yehuda report on a novel means by which nutrient-starved B. subtilis cells escape from aging (days-old) colonies by accumulating mutations enabling them to continue growth under nutrient-limited conditions. They postulate that such a strategy may be a major factor determining the dynamics of bacterial populations in natural environments.

Blueberries as an additive to increase the survival of Lactobacillus johnsonii N6.2 to lyophilisation

Effective cultivation methods, total cost, and biomass preservation are key factors that have a significant impact on the commercialisation and effectiveness of probiotics, such as Lactobacillus. Sugar polymers, milk and whey proteins have been suggested as good additives for industrial preparations. Alternative compounds, such as phytophenols, are a more attractive option, given their potential benefits to human health. The overall goal of this study was to determine if the addition of blueberry phytophenols improves the survival of Lactobacillus johnsonii N6.2 during the freeze-drying process. The addition of blueberry aqueous extract (BAE) stimulated the growth of L. johnsonii under aerobic conditions and improved the stationary phase survival of the bacteria. Furthermore, the addition of BAE to the culture media improved the endurance of L. johnsonii N6.2 to freeze-drying stress, as well as to storage at 4 °C for up to 21 weeks. Moreover, blueberry extract performed more effectively as a lyophilising additive compared to skim milk and microencapsulation with whey protein/sodium alginate. In sum, this study demonstrates that BAE is an effective additive to increase the growth and survival of L. johnsonii N6.2 when added to the culture medium and/or used as a lyophilising preservative. Moreover, BAE or other polyphenols sources might likely enhance growth and increase survival of more probiotic lactic acid bacterial strains.

Structural and functional insights into the stationary-phase survival protein SurE, an important virulence factor ofBrucella abortus

The stationary-phase survival protein SurE fromBrucella abortus(BaSurE) is a metal-dependent phosphatase that is essential for the survival of this bacterium in the stationary phase of its life cycle. Here, BaSurE has been biochemically characterized and its crystal structure has been determined to a resolution of 1.9 Å. BaSurE was found to be a robust enzyme, showing activity over wide ranges of temperature and pH and with various phosphoester substrates. The active biomolecule is a tetramer and each monomer was found to consist of two domains: an N-terminal domain, which forms an approximate α + β fold, and a C-terminal domain that belongs to the α/β class. The active site lies at the junction of these two domains and was identified by the presence of conserved negatively charged residues and a bound Mg2+ion. Comparisons of BaSurE with its homologues have revealed both common features and differences in this class of enzymes. The number and arrangement of some of the equivalent secondary structures, which are seen to differ between BaSurE and its homologues, are responsible for a difference in the size of the active-site area and the overall oligomeric state of this enzyme in other organisms. As it is absent in mammals, it has the potential to be a drug target.