J. Votruba

Degradation of polychlorinated biphenyls by extracellular enzymes of Phanerochaete chrysosporium produced in a perforated plate bioreactor

The white rot fungus Phanerochaete chrysosporium was cultivated in a perforated plate bioreactor and the expression of activities of manganese-dependent peroxidase (MnP) and lignin peroxidase (LiP) was measured. Peak activities of the two enzymes were reached close to day 11 and therefore the cultivation was terminated on that day. Extracellular proteins were concentrated and both peroxidases separated by isoelectric focusing. Degradation of technical PCB mixtures containing low and highly chlorinated congeners (Delor 103 and Delor 106 as equivalents of Aroclor 1242 and Aroclor 1260, respectively) was performed using intact mycelium, crude extracellular liquid and enriched MnP and LiP. A decrease in PCB concentration caused by a 44-h treatment with mycelium (74% w/w for Delor 103 and 73% for Delor 106) or crude extracellular liquid (62% for Delor 103 and 58% for Delor 106) was observed. The degradation was not substrate-specific, because no significant differences between the respective degradation rates were observed with di-, tri-, tetra-, penta-, hexa-, hepta-, and octachlorinated congeners. In contrast, MnP and LiP isolated from the above-mentioned extracellular liquid did not catalyse any degradation.

Organic fertilization changes the response of mycelium of arbuscular mycorrhizal fungi and their sporulation to mineral NPK supply

The synergetic effect of organic (cow manure) and mineral fertilization on the development arbuscular mycorrhizal (AM) fungi was demonstateded. The length of AM mycelium and sporulation were used as sensitive markers of the physiological state of soil AM fungal population. In manured treatments, both parameters increased in proportion with increasing mineral fertilization. In unmanured soil, the opposite trend was observed for the length of AM hyphae, which decreased with increasing mineral fertilization. Correlation analysis showed the dependence of length of AM hyphae and sporulation on soil available phosphorus. The correlation was negative in soil with no mineral fertilization and positive in soil supplied with luxury doses of mineral fertilizer.

Long-term fertilization affects the abundance of saprotrophic microfungi degrading resistant forms of soil organic matter

The effect of mineral and organic fertilization on the occurrence of soil microorganisms was determined in a field experiment. The colony-forming unit counts of saprotrophic microfungi, when estimated on a silicate gel medium containing fulvic acid as a sole carbon source, increased significantly with increasing doses of mineral and organic fertilization. Partial correlation analysis indicated that, unlike bacteria and actinomycetes, microfungi utilizing fulvic acid were significantly associated with soil organic carbon. No significant effects on bacteria and microfungi counted on common microbiological media were observed but counts of actinomycetes increased in a manured soil extensively fertilized by a mineral fertilizer. Fulvic acid utilizing microfungi, which are associated with areas rich in organics, play possibly the main role in mineralization of resistant forms of soil organic matter.

Influence of soluble humic substances on the growth of Algæ and blue-green Algæ

A stimulatory effect of soluble humic acid and sodium humate of soft coal origin on the growth of green algæChlorella andScenedesmus was established. On the contrary, the growth of blue-green algæ was not affected by humic substances. For green microorganisms soluble humic substances act more as a fertilizer than a bacteriostatic substance. The possibility of application of soluble humic substances as selective fertilizer of algæ at the expense of blue-green algæ at concentrations up to 300 mg/L is discussed.

Separation of ergot alkaloids by adsorption on silicates

A mixture of ergot alkaloids (agroclavine, elymoclavine, chanoclavine, and chanoclavine aldehyde) was separated from the Claviceps purpureafermentation broth by adsorption on inorganic adsorbents containing silica. The uptake of alkaloids depended on the concentration of adsorbent and pH. The adsorption capacity for of inorganic materials increased with increasing content of inorganic oxides such as MgO and CaO in the adsorbent. Using statistical thermodynamics, a simple mathematical model describing the multicomponent adsorption equilibrium is proposed and a numerical method suitable for fast computer simulation of multicomponent adsorption was developed.

Effect of salinity on the formation of avermectins, odor compounds and fatty acids byStreptomyces avermitilis

Streptomyces avermitilis was cultivated at different concentrations (0–12%) of NaCl in the medium and the content of fatty acids, production of odorous compounds and avermectins were measured. With increasing medium salinity the content of monoenoic fatty acids, particularly palmitoleic acid, increased while the quantity of branched fatty acids dropped. The production of avermectin was constant up to 0.5% of the salt in the medium and then it strongly decreased, reaching zero at 2.5% salt. The biosynthesis of oxolones and geosmin differed: the content of oxolones increased with growing salinity whereas the production of geosmin dropped.

An expert system applied to the physiological analysis of early stage of beer fermentation

A fuzzy expert system was applied to the knowledge analysis of yeast physiology in the early stage of beer fermentation, when the wort was aerated. We used ergosterol and glycogen concentration in the wort as a suitable marker of physiological state of the cell population. The amount of both compounds influences the rate of fermentation, cell growth and the final taste of beer. The concentrations of ergosterol and glycogen including the number of cells can not be measured immediately during the relatively short aeration period, and incomplete experimental data are therefore found in laboratory logbooks. We therefore suggested that the fuzzy relation between the directly measurable dissolved oxygen concentration and the rate of ergosterol or glycogen formation should be identified and a fuzzy expert system should be used to analyze the behavior of the yeast.

Kinetics of soluble glucan production by Claviceps viridis

Among 18 tested strains ofClaviceps spp., 7 produced significant amounts of exocellular polysaccharide (EPS). The maximum production of EPS was found in fermentation broth ofClaviceps viridis. The kinetics of growth, substrate consumption, and EPS production in the batch, aerobic, submerged culture of this fungus were investigated in detail. The experimental data were processed by a simple mathematical model describing mass balance of growth, substrate consumption, formation of intermediates, and production of EPS. The parameters of the model were estimated from data obtained in cultivation performed in flasks and two laboratory fermentors of different size. Physiological similarity was obtained during process scale-up in volumetric ratio 1 : 100. The sugar consumption efficiency (52 %) and observed EPS productivity (1.9 kg/m3 per d) were comparable with literature data.

In vivo and In vitro function of the intracellular proteolytic apparatus in nongrowing bacillus megaterium under heat stress

Abstract. In Bacillus megaterium sporulating at 35°C, up to 90% of 10-min pulse-labeled proteins were degraded. Degradation proceeded in two waves. Short-lived proteins, i.e., intrinsically labile proteins and proteins made short-lived because of starvation, were mostly degraded during the reversible sporulation phase. Their amount corresponded to 20% or slightly more during 2 h. The second wave of protein degradation, which followed during the irreversible sporulation phase at 35°C, increased the amount of total degradable pulse-labeled proteins to about 90%. This wave was absent in the isogenic asporogenic mutant 27-36 or in the wild strain, whose sporulation was inhibited by increased temperature. The proportion of degradable proteins was thus reduced to less than 40% in the asporogenic mutant incubated at 35°C and to 46% in the wild strain whose sporulation was suppressed by the temperature of 47°C. Unlike sporulating cells, these cells were thus capable of degrading short-lived and denatured proteins, but were not able to degrade most of other proteins. The in vitro protein degradation was substantially enhanced by increasing the Ca2+ concentration, suggesting a role of Ca2+-dependent proteinase(s) in the process.

External conditions influencing ethanol oxidation by resting cells ofCandida utilis

Effects of ethanol inhibition, initial pH and buffering capacity of media on the catabolic activity of nongrowing cells ofCandida utilis were studied. Effects of external conditions on the kinetic of ethanol oxidation and cell respiration are described by mathematical models. The results revealed a significant influence of both the external pH and the buffering capacity of the medium on the kinetic parameters of catabolic activity. The inhibitory effect results in the bottleneck of one of the reaction of the citrate cycle, glyoxylate cycle or electron transfer in a respiratory chain although the total rate of ethanol dissimilation increases under these conditions.