Saponaria officinalis saponins as a factor increasing permeability of Candida yeasts' biomembrane.

Saponins are a large group of compounds, produced mostly by plants as a side product of their metabolic activity. These compounds have attracted much attention over the years mostly because of their surface activity and antibacterial, anti-inflammatory and antifungal properties. On the other hand, most of the hitherto research has concerned the action of saponins against microbial cells as a whole. Therefore, knowing the possible interaction of saponins with biomembrane, we decided to check in-vitro the influence of saponin-rich extract of Saponaria officinalis on spheroplasts of two Candida sp. The obtained results show that 10 mg L- 1 of extract increased the permeability of spheroplasts up to 21.76% relative to that of the control sample. Moreover, the evaluation of surface potential has revealed a decrease by almost 10 mV relative to that of the untreated samples. Such results suggest its direct correlation to integration of saponins into the biomembrane structure. The obtained results have proved the antifungal potential of saponins and their ability of permeabilization of cells. This proves the high potential of saponins use as additives to antifungal pharmaceutics, which is expected to lead to improvement of their action or reduction of required dosage.

Thermodynamic Consideration of the Solid Saponin Extract Drop-Air System.

The aim of this research was to elucidate the surface active properties of Saponaria officinalis (soapwort) extract containing the plant surfactants saponins. To this end, the advancing contact angle (θ) of water, formamide and diiodomethane on the glass, as well as θ of the aqueous solution of S. officinalis extract fractions on PTFE, PMMA and glass, were studied. Based on the obtained results, the wetting behaviour of saponins was considered with regard to the surface tension components and parameters of the solutions and solids. The investigations also involved the description of the θ isotherms, the dependences between the cosine of contact angle and/or the adhesion of the solution to the solid surfaces and solution surface tension, as well as the critical surface tension of PTFE, PMMA and glass wetting. These dependences were studied based on the saponin adsorption at the different interfaces, which was deduced from the dependence between the adhesion and surface tension of the solution, as well as using the Gibbs and Frumkin isotherm equations. This proved that the saponins are poor wetting agents and that the contact angle isotherm can be described by the exponential function of the second order as well as the Szyszkowski equation, but only for PTFE.

Co-interaction of nitrofuran antibiotics and the saponin-rich extract on gram-negative bacteria and colon epithelial cells.

Large-scale use of nitrofurans is associated with a number of risks related to a growing resistance to these compounds and the toxic effects following from their increasing presence in wastewater and the environment. The aim of the study was to investigate an impact of natural surfactant, saponins from Sapindus mukorossi, on antimicrobial properties of nitrofuran antibiotics. Measurements of bacterial metabolic activity indicated a synergistic bactericidal effect in samples with nitrofurantoin or furazolidone, to which saponins were added. Their addition led to more than 50% greater reduction in viable cells than in the samples without saponins. On the other hand, no toxic effect against human colon epithelial cell was observed. It was found that exposure to antibiotics and surfactants caused the cell membranes to be dominated by branched fatty acids. Moreover, the presence of saponins reduced the hydrophobicity of the cell surface making them almost completely hydrophilic. The results have confirmed a high affinity of saponins to the cells of Pseudomonas strains. Their beneficial synergistic effect on the action of antibiotics from the nitrofuran group was also demonstrated. This result opens promising prospects for the use of saponins from S. mukorossi as an adjuvant to reduce the emission of antibiotics into the environment.

3D printed polylactide scaffolding for laccase immobilization to improve enzyme stability and estrogen removal from wastewater.

This study reports a biocatalytic system of immobilized laccase and 3D printed open-structure biopolymer scaffoldings. The scaffoldings were computer-designed and 3D printed using polylactide (PLA) filament. The immobilization of laccase onto the 3D printed PLA scaffolds were optimized with regard to pH, enzyme concentration, and immobilization time. Laccase immobilization resulted in a small reduction in reactivity (in terms of Michaelis constant and maximum reaction rate) but led to significant improvement in chemical and thermal stability. After 20 days of storage, the immobilized and free laccase showed 80% and 35% retention of the initial enzymatic activity, respectively. The immobilized laccase on 3D printed PLA scaffolds achieved 10% improvement in the removal of estrogens from real wastewater as compared to free laccase and showed the significant reusability potential. Results here are promising but also highlight the need for further study to improve enzymatic activity and reusability.

Study of Interactions between Saponin Biosurfactant and Model Biological Membranes: Phospholipid Monolayers and Liposomes.

The aim of this study was to determine the effect of saponins-rich plant extract on two model biological membranes: phospholipid monolayers and liposomes. The Langmuir monolayer technique was used to study the interactions of model phospholipid membranes with saponins. The π-A isotherms were determined for DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine) monolayer with the addition of various concentrations of licorice saponins extracts and subjected to qualitative as well as quantitative analysis. Additionally, relaxation studies of the obtained monolayers were carried out and morphological changes were examined using Brewster angle microscopy. Moreover, changes in the structure of phospholipid vesicles treated with solutions of saponins-rich plant extracts were assessed using the FTIR technique. The size and zeta potential of the liposomes were estimated based on DLS methods. The obtained results indicated that the saponins interact with the phospholipid membrane formed by DPPE molecules and that the stability of the mixed DPPE/saponins monolayer strongly depends on the presence of impurities in saponins. Furthermore, it was found that the plant extract rich in saponins biosurfactant interacts mainly with the hydrophilic part of liposomes.

New Biocomposite Electrospun Fiber/Alginate Hydrogel for Probiotic Bacteria Immobilization.

Biotechnological use of probiotic microorganisms involves providing them with appropriate conditions for growth, but also protection against environmental changes caused by an exchange of the medium, isolation of metabolites, etc. Therefore, the research on effective immobilization of probiotic microorganisms should be focused in this direction. The present study aimed to evaluate the effectiveness of an innovative hybrid immobilization system based on electrospun nanofibers and alginate hydrogel. The analyses carried out included the study of properties of the initial components, the evaluation of the degree and durability of cell immobilization in the final material, and their survival under stress conditions. Effective binding of microorganisms to the hydrogel and nanofibers was confirmed, and the collected results proved that the proposed biocomposite is an efficient method of cell protection. In addition, it was shown that immobilization on electrospun nanofibers leads to the preservation of the highest cell activity and the least cell growth restriction as compared to free or lyophilized cells only. The completed research opens new perspectives for the effective immobilization of microorganisms of significant economic importance.

Evaluation of the physico-chemical properties of hydrocarbons-exposed bacterial biomass.

In the efforts for the removal of hazardous materials from the environment biological processes are a valuable tool. Although much attention has been paid to the changes in bacteria at the omics level, another, physical-chemical perspective on the issue is essential, as little is known of microbial response to continuous exposition on harmful substances. This study provides in-depth characterization of the physical-chemical parameters of bacterial biomass after hydrocarbons exposure. To provide comparability of the harmful effects of chlorotoluenes and xylenes non-exposed and 12-months hydrocarbons exposed cells were analyzed, using the advanced spectrometric methods, inverse gas chromatography and low-temperature N2 sorption to evaluate acid-base as well as dispersive properties of the studied biomass. Presented results indicate P. fluorescens B01 cells strategy aimed at protecting the cell, thus lowering its' biodegradation efficiency as a result of metabolic stress. The outcome of the study was that prolonged exposure to pollutants might reduce the bioavailability of hydrocarbons to bacteria cells, and consequently decrease the effectiveness of decontamination of polluted sites by indigenous microorganisms.

Electrospun poly(methyl methacrylate)/polyaniline fibres as a support for laccase immobilisation and use in dye decolourisation.

Novel electrospun poly(methyl methacrylate)/polyaniline electrospun fibres were produced, characterised, modified, and used as a support for laccase immobilisation by two methods: adsorption and covalent binding. Effective deposition of laccase by both methods was confirmed by FTIR and CLSM results. Nevertheless, the main objective of the study was to select the most favourable immobilisation conditions and prepare heterogeneous biocatalysts with the best possible catalytic properties. The highest relative activity of enzymes immobilised by adsorption and covalent binding were obtained after 1 h of immobilisation using laccase solution at a concentration of 1 mg/mL, at pH 5 and 25 °C. It was found that the immobilised enzymes, which were present in amounts of 110 mg/g and 185 mg/g for systems with adsorbed and covalently bonded laccase respectively, exhibited slightly lower substrate affinity, and in consequence also a lower maximum reaction rate, than the free enzyme. The stability of laccase improved significantly upon immobilisation: both heterogeneous biocatalysts retained over 80% relative activity even after 10 repeated catalytic cycles and 30 days of storage. The obtained systems were used for decolourisation of Remazol Brilliant Blue R dye from a model aqueous solution, resulting in removal efficiencies of 87% and 58% using adsorbed and covalently bonded laccase, respectively. The described approach to the removal of textile dye from model solution is significant for the sustainable and environmentally friendly decolourisation of various compounds from wastewater.