The structure of a resuscitation-promoting factor domain from Mycobacterium tuberculosis shows homology to lysozymes

Resuscitation-promoting factor (RPF) proteins reactivate stationary-phase cultures of (G+C)-rich Gram-positive bacteria including the causative agent of tuberculosis, Mycobacterium tuberculosis. We report the solution structure of the RPF domain from M. tuberculosis Rv1009 (RpfB) solved by heteronuclear multidimensional NMR. Structural homology with various glycoside hydrolases suggested that RpfB cleaved oligosaccharides. Biochemical studies indicate that a conserved active site glutamate is important for resuscitation activity. These data, as well as the presence of a clear binding pocket for a large molecule, indicate that oligosaccharide cleavage is probably the signal for revival from dormancy

Cell-wall synthesis and ribosome maturation are co-regulated by an RNA switch in Mycobacterium tuberculosis

The success of Mycobacterium tuberculosis relies on the ability to switch between active growth and non-replicating persistence, associated with latent TB infection. Resuscitation promoting factors (Rpfs) are essential for the transition between these states. Rpf expression is tightly regulated as these enzymes are able to degrade the cell wall, and hence potentially lethal to the bacterium itself. We have identified a regulatory element in the 5' untranslated region (UTR) of rpfB. We demonstrate that this element is a transcriptionally regulated RNA switch/riboswitch candidate, which appears to be restricted to pathogenic mycobacteria, suggesting a role in virulence. We have used translation start site mapping to re-annotate the RpfB start codon and identified and validated a ribosome binding site that is likely to be targeted by an rpfB antisense RNA. Finally, we show that rpfB is co-transcribed with ksgA and ispE downstream. ksgA encodes a universally conserved methyltransferase involved in ribosome maturation and ispE encodes an essential kinase involved in cell wall synthesis. This arrangement implies co-regulation of resuscitation, cell wall synthesis and ribosome maturation via the RNA switch

Biotechnology can Improve a Traditional Product as Table Olives

Table olives are fermented vegetables very popular in the world and especially in the Mediterranean countries. Five main styles (Spanish or Sevillian, Castelvetrano, Siciliano, Californian, and Greek) are diffused to produce commercial products, beside several traditional styles. Although the main preparation methods of table olives are known for a long time, they are not yet optimized systems, and each of them is characterized by advantages and disadvantages. The use of NaOH for green olive debittering is responsible for the elimination of many aroma compounds and nutritionally important molecules. High volumes of heavily contaminated wastewaters are produced during olive processing. Spontaneous fermentation processes used to ferment black or green olives are difficult either to monitor or control. Microbial starters, selected for specific bio/technological and safety traits, can be useful to (i) improve the table olives organoleptic characteristics, (ii) control the fermentation process and significantly reduce the time to obtain a final product, (iii) monitor the correct evolution of the process, (iv) ensure the maintenance and/or improvement of nutritional and healthy features of the product, (v) protect table olives from undesired spoilage and pathogenic microorganisms, (vi) produce table olives as a carrier of microorganisms with probiotics characters, and (vii) enhance product stability and shelf life

Cascade oxime formation, cyclization to a nitrone, and intermolecular dipolar cycloaddition.

Simple haloaldehydes, including enolisable aldehydes, were found to be suitable for the formation of cyclic products by cascade (domino) condensation, cyclisation, dipolar cycloaddition chemistry. This multi-component reaction approach to heterocyclic compounds was explored by using hydroxylamine, a selection of aldehydes, and a selection of activated dipolarophiles. Initial condensation gives intermediate oximes that undergo cyclisation with displacement of halide to give intermediate nitrones; these nitrones undergo in situ intermolecular dipolar cycloaddition reactions to give isoxazolidines. The cycloadducts from using dimethyl fumarate were treated with zinc/acetic acid to give lactam products and this provides an easy way to prepare pyrrolizinones, indolizinones, and pyrrolo[2,1-a]isoquinolinones. The chemistry is illustrated with a very short synthesis of the pyrrolizidine alkaloid macronecine and a formal synthesis of petasinecine

Novel Approaches in Food Preservation and Their Impact on the Food System

: The increase in consumer demand for safe, convenient, and fresh food with an extended shelf life is accompanied by an interest in the environmental impacts caused by the food industry [...]

Selection of indigenous yeast strains for the production of sparkling wines from native Apulian grape varieties

Abstract We report the first polyphasic characterization of native Saccharomyces cerevisiae in order to select candidate strains for the design of starter cultures tailored for Apulian sparkling wines obtained from local grape variety. In addition, it is the first survey in our region that propose the selection of autochthonous starter cultures for sparkling wine i) including a preliminary tailored genotypic and technological screening, and ii) monitoring analytical contribution during secondary fermentation in terms of volatile compounds (VOCs). Furthermore, we exploit the potential contribute of autochthonous cultures throughout the productive chain, including the possible improvement of base wine. One representative strain from each cluster was characterized i) for tolerance to abiotic and biotic stressors peculiar of sparkling wine fermentation, ii) for the performances in base wine production, and iii) for the aptitudes to promote in-bottle secondary fermentation in white and rose sparkling wines, both obtained from Apulian grape varieties. Genetic characterization led to group 164 S. cerevisiae in 16 genetic clusters based on interdelta profiles. Stress tolerance assays shown a certain correlation with fermentative attitude. Our evidences demonstrated a different fermentative behavior and release of VOCs of the different strains in association with primary and secondary fermentations and as function of wine and rose sparkling wine. Furthermore, performances in white/rose sparkling wines have been found to be strain-dependent characters. Overall, we propose different strains as biotechnological resources suitable to improve the quality of regional sparkling wines and to provide a driver of innovation/segmentation in the market

Two-pronged attack: dual inhibition of Plasmodium falciparum M1 and M17 metalloaminopeptidases by a novel series of hydroxamic acid-based inhibitors

Plasmodium parasites, the causative agents of malaria, have developed resistance to most of our current antimalarial therapies, including artemisinin combination therapies which are widely described as our last line of defense. Antimalarial agents with a novel mode of action are urgently required. Two Plasmodium falciparum aminopeptidases, PfA-M1 and PfA-M17, play crucial roles in the erythrocytic stage of infection and have been validated as potential antimalarial targets. Using compound-bound crystal structures of both enzymes, we have used a structure-guided approach to develop a novel series of inhibitors capable of potent inhibition of both PfA-M1 and PfA-M17 activity and parasite growth in culture. Herein we describe the design, synthesis, and evaluation of a series of hydroxamic acid-based inhibitors and demonstrate the compounds to be exciting new leads for the development of novel antimalarial therapeutics

Effect of soil management and training system on negroamaro wine aroma

This study aimed to assess the impact of two soil managements and training systems on yield and wine aroma compounds of Negroamaro variety grown in a warm climate region (southern Italy). Cover crop (CC) and soil tillage (ST) as soil management, whilst bilateral Guyot (BG) and monolateral Guyot (MG) as training systems were compared. Free and bound volatile fractions were evaluated by GC-MS. ST and CC as well as BG and MG significantly affected yield parameters. In particular, yield was higher in ST and BG than in CC and MG, respectively; moreover, it was found to be positively influenced by interaction between BG and ST. Regarding aroma compounds, significant interactions between soil management and training system factors were observed. In case of free volatiles, the most positive interaction was found between BG and ST, whereas, for bound volatiles, the best interaction was represented by MG with both soil tillage and cover crop. Vine leaf area and development over vine growth stages along with water stress levels played an important role in determining the aroma profile as well as yield parameters. In conclusion, the training system significantly interacted with soil management and affected most of important aroma compounds in Negroamaro wine

Complexity and dynamics of the winemaking bacterial communities in berries, musts, and wines from apulian grape cultivars through time and space

Currently, there is very little information available regarding the microbiome associated with the wine production chain. Here, we used an amplicon sequencing approach based on high-throughput sequencing (HTS) to obtain a comprehensive assessment of the bacterial community associated with the production of three Apulian red wines, from grape to final product. The relationships among grape variety, the microbial community, and fermentation was investigated. Moreover, the winery microbiota was evaluated compared to the autochthonous species in vineyards that persist until the end of the winemaking process. The analysis highlighted the remarkable dynamics within the microbial communities during fermentation. A common microbial core shared among the examined wine varieties was observed, and the unique taxonomic signature of each wine appellation was revealed. New species belonging to the genus Halomonas were also reported. This study demonstrates the potential of this metagenomic approach, supported by optimized protocols, for identifying the biodiversity of the wine supply chain. The developed experimental pipeline offers new prospects for other research fields in which a comprehensive view of microbial community complexity and dynamics is desirable.Peer ReviewedPostprint (published version