Advanced characterization of natural biofilm on nanofiber scaffold.

Nanofiber scaffolds provide numerous advantages over common carriers engineered for microorganisms. The most important advantage is an increased speed of primary surface colonization (up to four times faster), which shortens the time required for the areal biofilm formation and optimum performance of attached microorganisms (higher efficiency of biological activity of up to twice as fast). Image analysis predicts early formation of biofilm even in beginning stages; analysis of biofilm reveals the different structures of bacterial colonies on both scaffolds (higher porosity, size, and number of bacterial colonies on nanofiber's surface). The image analysis correlates well with determinations of dry matter (linear correlation of 0.96) and proteins (linear correlation of 0.89).

Mild hydrolysis of nitriles by Fusarium solani strain O1.

High levels of an aromatic nitrilase (about 37 microkat/L culture) were induced in Fusarium solani O1 after transfer of the mycelium from a rich medium into a medium with 20 mmol/L picolinonitrile. The mycelium was entrapped in lense-shaped particles consisting of a polyvinyl alcohol/polyethylene glycol copolymer (LentiKats). The cell-free extract was immobilized by hydrophobic binding onto a Butyl Sepharose column. The enzyme was useful for the mild hydrolysis of nicotinonitrile, isonicotinonitrile and benzonitrile.

Colonization of surfaces by phenolic compounds utilizing microorganisms.

The aim of the present study is to determine optimal adhesive interaction of phenolic compounds utilizing Candida maltosa and Rhodococcus erythropolis when adhering to kaolin, silicone, synthetic foil (Steriking R40) and fluorinated silicones, comparing cell and support surface hydrophobicity. In parallel, the interfering effect of detergents was investigated. Data obtained show that the less hydrophobic supports display high initial cell adhesion when contacted with the cell type with a lower surface hydrophobicity (yeast cell) but most stable yeast biofilms are those formed on highly hydrophobic fluorinated silicones. On the other hand, support hydrophobicity has no effect on bacterial cell detachment; however, bacterial biofilms are denser when growing on more hydrophobic supports. Both detergents interfere (independently on the cell type) with the early and late phases of biofilm development.

The influence of heavy metals on the production of extracellular polymer substances in the processes of heavy metal ions elimination.

Wastewaters from a chemical industry polluted by heavy metal ions represent a hazard for all living organisms. It can mean danger for ecosystems and human health. New methods are sought alternative to traditional chemical and physical processes. Active elimination process of heavy metals ions provided by living cells, their components and extracellular products represents a potential way of separating toxic heavy metals from industrial wastewaters. While the abilities of bacteria to remove metal ions in solution are extensively used, fungi have been recognized as a promising kind of low-cost adsorbents for removal of heavy-metal ions from aqueous waste sources. Yeasts and fungi differ from each other in their constitution and in their abilities to produce variety of extracellular polymeric substances (EPS) with different mechanisms of metal interactions. The accumulation of Cd(2+), Cr(6+), Pb(2+), Ni(2+) and Zn(2+) by yeasts and their EPS was screened at twelve different yeast species in microcultivation system Bioscreen C and in the shaking Erlenmayer's flasks. This results were compared with the production of yeast EPS and the composition of yeast cell walls. The EPS production was measured during the yeast growth and cell wall composition was studied during the cultivations in the shaking flasks. At the end of the process extracellular polymers and their chemical composition were isolated and amount of bound heavy metals was characterized. The variable composition and the amount of the EPS were found at various yeast strains. It was influenced by various compositions of growth medium and also by various concentrations of heavy metals. It is evident, that the amount of bound heavy metals was different. The work reviews the possibilities of usage of various yeast EPS and components of cell walls in the elimination processes of heavy metal ions. Further the structure and properties of yeasts cell wall and EPS were discussed. The finding of mechanisms mentioned above is necessary to identify the functional groups entered in the metals elimination processes.

Host-vector system for phenol-degrading Rhodococcus erythropolis based on Corynebacterium plasmids.

The strain Rhodococcus erythropolis CCM2595, which was shown to degrade phenol, was chosen for genetic studies. To facilitate strain improvement using the methods of gene manipulation, the technique of genetic transfer was introduced and cloning vectors were constructed. Using the plasmid pFAJ2574, an electrotransformation procedure yielding up to 7x10(4) transformants/microg DNA was optimized. Escherichia coli- R. erythropolis shuttle vectors were constructed using the replicons pSR1 and pGA1 from Corynebacterium glutamicum. The small vector pSRK21 (5.8 kb) provides six unique cloning sites and selection of recombinant clones using alpha-complementation of beta-galactosidase in E. coli. This vector, exhibiting high segregational stability under non-selective conditions in R. erythropolis CCM2595, was applied to cloning and efficient expression of the gene coding for green fluorescent protein (gfpuv).

Comparison of yeast (Candida maltosa) and bacterial (Rhodococcus erythropolis) phenol hydroxylase activity and its properties in the phenolic compounds biodegradation.

Aromatic contaminants of the environment, to which belongs phenol and its derivatives, are toxic and in most of the cases hard to degrade. Removal of these pollutants by biological, gentle and effective way, depends on specific environmental conditions in the locality and on the biodegradation potential of the used microbial population. Closer characterization of the biodegradation and enzyme mechanisms is therefore an essential assumption of the successful implementation of microbes. This paper is focused on comparison of the biodegradation activity between the soil yeast Candida maltosa and bacteria Rhodococcus erythropolis towards various aromatics connected with determination of the first enzyme of the phenol biodegradation pathway: phenol hydroxylase (PH). The effect of substrate type, substrate concentration, growth phase of the microorganisms and presence of humic acids in the cultivation medium, on phenol biodegradation and PH activity are discussed.

The enhancement of reproduction and biodegradation activity of eukaryiotic cells by humic acids.

Fourteen samples of humic acids (HA) were screened for ability to influence reproduction and biodegradation activity of eukaryotic cells in the presence of chosen toxic pollutants. Microorganisms Candida maltosa and Rhodotorula mucilaginosa (soil isolates) were used for all tests. It was observed during our experiments that some samples of humic acids served as a protection against the high concentration of toxic pollutants (phenol, naphtalene etc). This effect can be widely used in many bioremediation technologies.

Significance of physical attachment of fungi for bio-treatment of water.

The inhibitory effect of xenobiotics known to damage cell surface structures was studied. The sensitivity of suspended cells of the two fungi Candida maltosa and Fusarium proliferatum was compared with that of artificial or natural biofilms of these fungi. The results obtained indicate that the resistance of attached cell populations to model xenobiotics is increased compared with suspended cells. Only the attached fungal cells had the capacity to degrade acetone and phenol and to adapt to increasing concentrations of these substances, so they seem ideally suited for bioremediation of waste water.

[A rare case of necrotizing fasciitis and subsequent sepsis with multiorgan failure caused by Vibrio vulnificus]. / Vzácný prípad nekrotizující fasciitidy a následné sepse s multiorgánovým selháním zpusobený agens Vibrio vulnificus.

The authors report a rare case of necrotizing fasciitis, sepsis caused by a strain of Vibrio vulnificus rare in Czech Republic.

Changes in the starvation response through covalent cell attachment.

Covalent attachment of Candida utilis cells, possibly simulating natural microbial immobilizations, stimulated stable and significant enhancement of extracellular production of alkaline protease, specifically induced by four different starvation conditions. The enzyme analysis confirmed the identity of the proteases released under all conditions of starvation and no parallel production of other proteolytic enzyme. The enhancement phenomenon as a uniform and stable effect of the whole cell immobilization is discussed in relation to the effect of multipoint, cell-solid surface contact, potentially bringing positive modulations of complex, cellular functions.

Use of mineral nutrients and surface-active substances in a biodegradation process modulation.

The capability of a bacterial population to degrade oil hydrocarbons and naphthalene was found to be markedly enhanced by an optimized P:N ratio as well as by proper application of a surface-active compound. The importance of this optimization procedure was shown by both laboratory and technologically performed experiments.

Covalent immobilization as a stimulus of cell wall composition changes.

Covalent immobilization of yeast cells by an activated diamine spacer is accompanied by increased levels of cell wall proteins, lipids, amino sugars, amino acids and acid phosphatase leakage, and by altered composition of mannoproteins. The observed changes in cell wall composition are attributed to the effect of cell-solid surface contact.

Novel way to study the chronological changes in yeast outer-wall proteins.

The time course of changes in the capacity of synchronized yeast cells to be covalently immobilized was determined and interpreted as an indication of chronological changes in the cell's outer-wall proteins. Corroborative evidence was obtained indicating that these transient changes are dependent on protein synthesis and connected with the progression of the cell cycle through the late S and/or early G2 phases.

Nystatin and killer toxin sensitivity of free and immobilizedSaccharomyces cerevisiae.

The inhibitory effect of nystatin and killer toxin on the growth of free and covalently-immobilizedSaccharomyces cerevisiae cells was studied. The resistance of immobilized cells to both agents was accompanied by increased amounts of phospholipids and sterols. The possible relationship between these changes in the membrane composition and the transduction of a signal across the cytoplasmic membrane is discussed.

Quantitative changes of wall polysaccharides in an immobilized yeast.

Whole cells of Saccharomyces cerevisiae were covalently linked to modified polyphenyleneoxide. The immobilization was accompanied by retention of cell division and increased concentrations of wall alkali-insoluble glucans and mannan.

Immobilized cells.

Three basic types of immobilization (i.e. without carrier, entrapment and immobilization on the carrier surface) of microbial cells, nonmicrobial cell populations and subcellular organelles are reviewed. These are further developed into a number of combined and less frequently used techniques of immobilization and application of cell biocatalysts for industrial biotransformation in pharmacy, food industry and agriculture, including novel approached and some unpublished authors' results.

Inhibitory effect of 5-bromo-6-azauracil on growth and cell division of Saccharomyces cerevisiae.

The most marked effect of 5-bromo-6-azauracil (BrAzU) on yeast cells is to cause cell lysis. The inhibition of the lytic process is delayed and this delay coincides in time with the capacity of preformed pyrimidines to reverse the effect of BrAzU.