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Microbial extradiol dioxygenases have a great potential in bioremediation, but their structure is very sensitive to various environmental and chemical agents. Immobilization techniques make the enzyme properties’ improvement possible. This is the first report of the usage of κ-carrageenan as a matrix for the immobilization of catechol 2,3-dioxygenase. The storage stability of entrapped catechol 2,3-dioxygenase from Stenotrophomonas maltophilia KB2 in κ-carrageenan hydrogel at 4ºC was found up to 14 days, while the free enzyme lost its activity within 24 hours. The immobilization of dioxygenase decreased the optimum temperature by 10ºC, while both soluble and immobilized enzyme showed similar pH properties. The Km, Vmax, and Hill constant values for the immobilized enzyme were 0.17 μM, 106.68 mU, and 1.00, respectively. The immobilized catechol 2,3-dioxygenase showed higher activity against 3-methylcatechol, hydroquinone, and tetrachlorohydroquinone than the soluble enzyme. Immobilization of catechol 2,3-dioxygenase protected the enzyme from inhibition and enhanced its resistance to inactivation during catalysis.
For L-aspartic acid biosynthesis, high production cells of Escherichia coli mutant B-715 and P1 were immobilized in chitosan gel using a technique developed in our laboratory. The immobilization process reduced initial activity of the intact cells, however, the biocatalyst produced was very stabile for long-term use in multi-repeated batch or continuous processes. Temperature influence on the conversion of ammonium fumarate to L-aspartic acid was investigated. In long-term experiments, over 603 hours, the temperature 40°C was found to be the best for both biocatalyst stability and high conversion rate. The optimum substrate concentration was 1.0 M. Continuous production of L-aspartic acid was investigated in three types of column bioreactors characterized by different volumes as well as different high to biocatalyst bed volume rations (Hz/Vz). The highest conversion rate, 99.8%, and the productivity 6 g/g/h (mass of L-aspartic acid per dry mass of cells in biocatalyst per time unit) was achieved in the bioreactor with the highest value Hz/Vz = 3.1, and liquid hour space velocity value of 5.2, defined as the volume of feeding substrate passed per volume of catalyst in bioreactor per one hour.
The effect of soil contamination with diesel oil and petrol on the nitrification process was investigated in a laboratory experiment. Samples of typical brown soil developed from loamy sand, of pH of 6.6 in 1M KCl, Hh -11.38 mmol+ kg-1 soil, S — 77.67 mmol+ kg-1 soil and Corg - 8.50 g kg-1 were analyzed. The experiment was performed in three replications, and for each test 100 g air-dry soil sample was placed in 150 cm3 beakers. Soil samples were contaminated with diesel oil and petrol with the addition of rapeseed oil and ethanol. The source of nitrogen was ammonium sulfate in the amount of 0 and 250 mg N per kg-1 soil. The content of N-NO3- and N-NH4+ was determined on experimental days 14, 28 and 42. Soil moisture was kept constant at 50% capillary water capacity throughout the experiment. Fertilizer nitrogen was subject to strong immobilization in soil contaminated with diesel oil and petrol. Both pollutants strongly inhibited the nitrification process. Diesel oil had a much stronger inhibitory effect on nitrification than petrol. Rapeseed oil also proved to be a powerful inhibiting factor. On experimental day 42, diesel oil reduced ammonium cation oxidation by 99%, and petrol - by 88%.
This experiment was conducted in a bioreactor with biomass immobilized in ceramic carriers. The influence of hydraulic retention time (HRT), carrier structure and intrinsic circulation rate on carbon and nitrogen removal from municipal wastewater were investigated. Two types of ceramic carriers were used at HRT 70, 60, 40, 30 min for carrier I, and 70, 60, 30, 15 min for carrier II, and at the circulation rate of 60, 40 and 20 dm3·h-1. The highest nitrogen removal efficiency was achieved in carrier II at 30 min of reaction. The carbon removal efficiency was similar for both carriers. An increase in internal circulation rate from 20 to 60 dm3·h-1 enhanced nitrogen removal efficiency from 33.0 to 47.2% and decreased in the production of surplus sludge in carrier II.
We performed the ultrastructural evaluation of presence and of some basic morphometric parameters of nemaline structures (NS) and Z band malformations in the musculus soleus fibres of rats kept in 7 months lasting immobilisation. Disturbances in Z band, leading to NS formation, were observed very often. Also numerous, classically rod-shaped NS were found in analyzed material. We demonstrate, for the first time in literature, that long-lasting hypokinesia induces NS formation. The pictures obtained may suggest successive steps in NS formation and maturation.
Ethanol producing bacteria Zymomonas mobilis (strain 3881 and 3883) were used in batch and continuous fermentation as free cells as well as immobilized in alginate beads. Continuous fermentation helped to increase the productivity of fermentors significantly and continuous fermentation with immobilized in alginate cells gave as high productivities as 49,5 g/dm3*h.
The aim of this work was to study the influence of hypokinetic conditions on the ovary and corpus luteum of pregnant rats. The rats were kept in hypokinetic conditions for 5 days in the period between the 13th and 18th days of pregnancy. A three-dimensional reconstruction of the ovary and corpora lutea and also a stereological evaluation of the luteal cells and their nuclei were performed using serially cut material. Hypokinesia caused a decrease in the mean volume of the ovary and individual corpus luteum and in the total volume of corpora lutea per ovary in immobilised animals as compared to the control. Moreover, a decrease was observed in the mean number of luteal cells and an increase in the size of these cells, as well as in the mean volume fraction of their nuclei. These results indicate that immobilisation of pregnant rats for 5 days considerably influences the morphology of the corpus luteum and luteal cells.
This paper presents the results of laboratory research on the process of heavy metals extraction from sludge, where heavy metals content exceeds permissible levels. The method of immobilization of heavy metals depends on using the developed technology hygienization process of wastewater sludge (which is pending by patent) to environment utilization. The base of this technology is treatment of municipal wastewater sludge by dust from electrofilters of cement mills with the addition of roasted raw detrital basalt. The modification of this technology involves using the additional unit operation that is followed by extraction of the excessive amount of heavy metals from sludge. The results of heavy metals extraction from wastewater sludge were presented.
The conditions of red clover dressing are defined to a great extent by the fertility of sward-podzolic soils. Having soil with low level of fertility (pH 5.2; P₂O₅ – 110; K₂O – 120 mg/kg of soil) after harvesting cover crop (in the fall) it is necessary to apply P₆₀K₉₀. Top dressing during the phase of budding at the beginning of clover blossom (in spring) is done with microfertilizers containing molybdenum. On soils with average level of fertility (pH 5.7; P₂O₅ – 185; K₂O – 190 mg/kg of soil) after harvesting cover crop it is necessary to apply P₄₀K₆₀ to dress clover. Top dressing during the phase of budding at the beginning of clover blossom is done with microfertilizers containing boron and molybdenum. On soils with high level of fertility (pH 6.2; P₂O₅ – 300; K₂O – 260 mg/kg of soil) after harvesting cover crop it is necessary to apply P₄₀K₆₀. Top dressing during the phase of budding at the beginning of clover blossom is made with microfertilizers containing boron.
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