Biologia最新文献

Salmonella Typhi infection is showing as a negative alarm for human health due to the occurrence of life-threatening typhoid fever and the emergence of antibiotic resistance. The infection becomes chronic due to the biofilm formation of S. Typhi, which plays a significant role in antibiotic resistance. The S. Typhi can survive under various stress conditions such as antibiotic stress, host immune stress, environmental stress, and nutritional stress in the biofilm stage. Biofilm provides a protective cover to the S. Typhi and prevents the entry of antibiotics inside the bacterial cells. Thus, biofilm is the root cause of the problem. Therefore, this study explores the easy detection of S. Typhi biofilms using safranin dye with specific concentrations. We performed the biofilm growth kinetics, qualitative assessment of the biofilm formations, and safranin interactions to finalize the specific concentrations of safranin (0.5 mg/mL) to detect the S. Typhi biofilm condition. The study was validated at 0.5 mg/mL safranin concentrations with the help of RSM analysis and kinetics study of the typhoidal biofilm. The study is important because the obtained results indicated the precise detection of S. Typhi biofilm, which is useful for all phases of biofilm detection. The study provided new directions for typhoidal biofilm staining that could be an emerging methodology in the area of health sciences.

The black timber bark beetle (Xylosandrus germanus) is a strongly invasive ambrosia beetle and an important forest pest in Slovakia. This pest is closely associated with symbiotic fungi used as its food source. We investigated the fungi associated with X. germanus adults in Slovakia. In this study, Beauveria bassiana, B. pseudobassiana, Clonostachys rosea, Fusarium oxysporum, Ophiostoma quercus, Phaeoacremonium scolyti, and Talaromyces amestolkiae were isolated and identified by morphological and molecular analyses. The fungus Ophiostoma quercus was most frequently isolated from living beetles, while the entomopathogenic Beauveria bassiana was the most commonly isolated from dead beetles. The morphological descriptions of fungi based on isolates from the surface of X. germanus adults are provided.

N-glycosylation of recombinant proteins using bacterial glycosylation system has proven to be a valuable although developing tool ultimately applicable to various industries. When used for enzyme engineering, it offers the possibility of increased stability or immobilization route and thus increasing effectiveness of e.g. biotransformation or other biocatalysis procedures. One such promising enzyme is alcohol dehydrogenase (ADH) for use in redox biotransformation reactions. Given the current possibilities of recombinant enzyme production, including major advances in glycoengineering and glycoprotein production in bacterial organisms, the aim of this work was the production of thermotolerant ADH from Rhodococcus ruber (RrADH) in glycosylated form in Escherichia coli. We have successfully developed a dual plasmid expression system enabling glycosylation of target proteins utilizing a glyco-tag approach. We were able to produce RrADH in soluble form and at the same time we detected a bacterial glycan conjugated to RrADH as well as the activity of the enzyme. The glycan bound to recombinant enzyme can be used for oriented covalent immobilization of the enzyme, which would increase the potential for its practical application in biotransformation of various compounds.

In this work, we focus on the identification of novel fungal peroxygenase gene belonging to the peroxidase-peroxygenase superfamily. We applied a metagenomic approach on soil samples from primeval forest and appropriate bioinformatics tools for analysis of obtained genomic DNA sequence. Peroxidases are ubiquitous metalloenzymes that are able to reduce reactive peroxides, mainly hydrogen peroxide, into water, whereas several substrates can be concomitantly oxidized during their catalytic reaction. Our purpose was to collect unique peroxygenase sequence data originating from a preserved biotope for a robust phylogenetic reconstruction of a particular gene family coding for highly versatile heme-thiolate peroxidases that has peculiar yet undiscovered representatives among ectomycorrhizal fungi. We identified unique DNA sequence, 812 bp long, from ectomycorrhizal Suillus species coding for a heme-thiolate peroxidase with 1 typical intron that appears distinctive for Carpathian forests. After translation in corresponding protein sequence 251 amino acids long we could identify typical signatures of this peroxygenase. On the proximal side of heme we found the conserved P-C-P triad responsible for efficient ligation of heme iron thus influencing the reactivity of this peroxidase. On the distal side we recognized the E-H-D-X-S-L motif for interaction with a stabilizing magnesium ion. Maximum likelihood reconstruction of protein phylogeny revealed with a high bootstrap support the presence of a monophyletic HTP4 clade originating in numerous Suillus representatives. Together with sister clades of edible Boletus and poisonous Paxillus containing diverse peroxygenases these newly discovered heme catalyst can be considered for application of oxyfunctionalization of organic molecules.

Biostimulants such as ascorbic acid, known as vitamin C, have been reported to have numerous positive roles in plant tolerance to abiotic stresses. However, little is known about the biostimulant effects of ascorbic acid on alfalfa (Medicago sativa). Accordingly, a pot experiment was conducted to investigate the effects of 1 mM ascorbic acid, applied as foliar spray, on the salt tolerance of a Moroccan alfalfa population Demnate 201. One month-old M. sativa seedlings were exposed to 200 mM NaCl for four weeks with or without 1 mM of exogenous ascorbic acid treatment. The results showed that salinity stress significantly (p < 0.001) reduced plant biomass, disturbed photosynthesis-related parameters and induced oxidative stress. However, ascorbic acid foliar spray counteracted the observed negative effects of salinity. It significantly (p < 0.001) improved plant growth and photosynthetic parameters. Besides, stress indicators, including Na⁺ in shoot and root, hydrogen peroxide and electrolyte leakage, were significantly reduced by 42%, 29%, 12% and 34%, respectively, in treated and salt-stressed alfalfa plants. Interestingly, the decrease in oxidative stress markers was positively correlated to the ability of ascorbic acid to induce the accumulation of flavonoids and to increase the antioxidant activity of guaiacol peroxidase. Furthermore, compatible solutes, such as proline and soluble sugars, were found higher especially in salt-stressed alfalfa plants treated with 1 mM ascorbic acid. Our findings showed that ascorbic acid supply could be an eco-friendly and sustainable technique to mitigate the toxic effect of salt and could improve alfalfa forage production when grown in salt-affected soils.