Simultaneous Biodegradation of Phenol and n-Hexadecane by Cryogel Immobilized Biosurfactant Producing Strain Rhodococcus wratislawiensis BN38.

The capability of the biosurfactant-producing strain Rhodococcus wratislawiensis BN38 to mineralize both aromatic and aliphatic xenobiotics was proved. During semicontinuous cultivation 11 g/l phenol was completely degraded within 22 cycles by Rhodococcus free cells. Immobilization in a cryogel matrix was performed for the first time to enhance the biodegradation at multiple use. A stable simultaneous hydrocarbon biodegradation was achieved until the total depletion of 20 g/l phenol and 20 g/l n-hexadecane (40 cycles). The alkanotrophic strain R. wratislawiensis BN38 preferably degraded hexadecane rather than phenol. SEM revealed well preserved cells entrapped in the heterogeneous super-macroporous structure of the cryogel which allowed unhindered mass transfer of xenobiotics. The immobilized strain can be used in real conditions for the treatment of contaminated industrial waste water.

In Vitro Multiplication and Rooting of Plum Rootstock 'Saint Julien' (Prunus domestica subsp. insititia) under Fluorescent Light and Different LED Spectra.

In recent years, light emitting diodes (LEDs), due to their low energy consumption, low heat emission and specific wavelength irradiation, have become an alternative to fluorescent lamps (FLs) in plant tissue culture. The aim of this study was to investigate the effects of various LED light sources on the in vitro growth and rooting of plum rootstock Saint Julien (Prunus domestica subsp. insititia). The test plantlets were cultivated under a Philips GreenPower LEDs research module illumination system with four spectral regions: white (W), red (R), blue (B) and mixed (W:R:B:far-red = 1:1:1:1). The control plantlets were cultivated under fluorescent lamps (FL) and the photosynthetic photon flux density (PPFD) of all treatments was set at 87 ± 7.5 µmol m-2 s-1. The effect of light source on the selected physiological, biochemical and growth parameters of plantlets was monitored. Additionally, microscopic observations of leaf anatomy, leaf morphometric parameters and stomata characteristics were carried out. The results showed that the multiplication index (MI) varied from 8.3 (B) to 16.3 (R). The MI of plantlets grown under mixed light (WBR) was 9, lower compared to the control (FL) and white light (W), being 12.7 and 10.7, respectively. In addition, a mixed light (WBR) favored plantlets' stem growth and biomass accumulation at the multiplication stage. Considering these three indicators, we could conclude that under the mixed light, the microplants were of better quality and therefore mixed light (WBR) was more suitable during the multiplication phase. A reduction in both net photosynthesis rate and stomatal conductance in the leaves of plants grown under B were observed. The quantum yield (Yield = FV/FM), which represents the potential photochemical activity of PS II, ranged from 0.805 to 0.831 and corresponded to the typical photochemical activity (0.750-0.830) in the leaves of unstressed healthy plants. The red light had a beneficial effect on the rooting of plum plants; the rooting was over 98%, significantly higher than for the control (FL, 68%) and the mixed light (WBR, 19%). In conclusion, the mixed light (WBR) turned out to be the best choice during the multiplication phase and the red LED light was more suitable during the rooting stage.

Bacterial Natural Disaccharide (Trehalose Tetraester): Molecular Modeling and in Vitro Study of Anticancer Activity on Breast Cancer Cells.

Isolation and characterization of new biologically active substances affecting cancer cells is an important issue of fundamental research in biomedicine. Trehalose lipid was isolated from Rhodococcus wratislaviensis strain and purified by liquid chromatography. The effect of trehalose lipid on cell viability and migration, together with colony forming assays, were performed on two breast cancer (MCF7-low metastatic; MDA-MB231-high metastatic) and one "normal" (MCF10A) cell lines. Molecular modeling that details the structure of the neutral and anionic form (more stable at physiological pH) of the tetraester was carried out. The tentative sizes of the hydrophilic (7.5 Å) and hydrophobic (12.5 Å) portions of the molecule were also determined. Thus, the used trehalose lipid is supposed to interact as a single molecule. The changes in morphology, adhesion, viability, migration, and the possibility of forming colonies in cancer cell lines induced after treatment with trehalose lipid were found to be dose and time dependent. Based on the theoretical calculations, a possible mechanism of action and membrane asymmetry between outer and inner monolayers of the bilayer resulting in endosome formation were suggested. Initial data suggest a mechanism of antitumor activity of the purified trehalose lipid and its potential for biomedical application.

Characterization and potential antitumor effect of a heteropolysaccharide produced by the red alga Porphyridium sordidum.

Taking into account the rising trend of the incidence of cancers of various organs, effective therapies are urgently needed to control human malignancies. However, almost all chemotherapy drugs currently on the market cause serious side effects. Fortunately, several studies have shown that some non-toxic biological macromolecules, including algal polysaccharides, possess anti-cancer activities or can increase the efficacy of conventional chemotherapy drugs. Polysaccharides are characteristic secondary metabolites of many algae. The efficacy of polysaccharides on the normal and cancer cells is not well investigated, but our investigations proved a cell specific effect of a newly isolated extracellular polysaccharide from the red microalga Porphyridium sordidum. The investigated substance was composed of xylose:glucose and galactose:manose:rhamnose in a molar ratio of 1:0.52:0.44:0.31. Reversible electroporation has been exploited to increase the transport through the plasma membrane into the tested breast cancer tumor cells MCF-7 and MDA-MB231. Application of 75 µg/mL polysaccharide in combination with 200 V/cm electroporation induced 40% decrease in viability of MDA-MB231 cells and changes in cell morphology while control cells (MCF10A) remained with normal morphology and kept vitality.

Examination of Humicola lutea immobilized in sol-gel matrices: effective source of alpha-galactosidase.

Alpha-Galactosidase production by the fungus Humicola lutea 120-5 immobilized in a hybrid sol-gel matrix, consisting of tetraethylorthosilicate (TEOS) as a precursor and a mixture of polyethyleneglycol (PEG) and polyvinylalcohol (PVA), was investigated under semicontinuous shake flask cultivation and compared to the enzyme secretion by free cells. The influence of the carrier weight on the alpha-galactosidase biosynthesis in repeated batch experiments was followed. Best results were obtained with 2 g of the sol-gel particles per culture flask using 144-h runs. The growth behaviour of the immobilized mycelium during both the growth and productive phases was observed by scanning electron microscopy. The presence of abundant mycelial growth of intact hyphae correlated with a 2-fold higher enzyme activity compared to free cells. The obtained biocatalyst retained a high level of enzyme titer exceeding the activity of free cells during four cycles of operation (24 days). This result is confirmed by the micrographs showing the retained viability of the growing vegetative cells due to the protective role of the carrier.

Ultracytochemical localization of acid phosphatase in Humicola lutea conidia and mycelia.

Electron microscopic cytochemical procedures were used to determine the cellular location of acid phosphatase in the fungus Humicola lutea grown in casein-containing medium lacking in mineral orthophosphates. In our investigations acid phosphatase in nongerminating conidia was localized on the outer side of the cell wall, in the cell wall, and on the exterior surface of the plasma membrane. The reaction product of acid phosphatase in germinating conidia was seen in the outer wall layer while in young mycelium on the cell surface and in the exocellular space. The relationship between phosphatase activities localized in the cell wall and their role in the enzymatic degradation of the phosphoprotein casein providing available phosphates for cell growth is discussed.