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Study of enzymes secreted by oomycete Pythium oligandrum
Hrdinová, Karolína ; Ryšlavá, Helena (advisor) ; Kubíčková, Božena (referee)
Pythium oligandrum is one of the non-pathogenic microorganisms of Pythium species which found its use as biological control agent. The main modes of action of this oomycete lie in inducing the plant immunity and in mycoparasitism of pathogenic fungi and fungus-like organisms. The oomycete attacks the pathogens by secretion of hydrolytic enzymes into the environment. In the first part of this bachelor's thesis, activity of hydrolytic enzymes endo-1,3-β-glucanase, cellulase, chitinase and proteases was observed in a commercial product based on Pythium oligandrum, called Polyversum-Biogarden. It was conclu- ded that the direct hydrolysis of phytopathogens is probably not the main mechanism of this product because the activity of glycosidases increased only after six hour-long incubation of the Polyversum-Biogarden in water and the proteolytic activity was not detected. In the second part of the bachelor's thesis, properties of proteases secreted by orga- nism Pythium oligandrum were studied. The highest proteolytic activity was observed at pH 6,5. Ovomucoid acted as an inhibitor of secreted proteases. The stability of pro- teases was lowered by SDS, detergents present in liquid soaps and by a solution of a solid soap or NaOH. Only the highest concentrations of urea lowered the activity of proteases....
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Immobilization of protein macromolecules onto polymer carriers: An overview
Badalcová, Helena ; Holas, Ondřej (advisor) ; Svačinová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of: Pharmaceutical Technology Consultant: PharmDr. Ondřej Holas, Ph.D. Student: Helena Badalcová Title of Thesis: Immobilization of protein macromolecules onto polymer carriers: An overview Since the 70s, the immobilised enzymes have been getting the attention of not only scientific and laboratory workers, but also industrial companies. Enzymes are unique biocatalysts, which are distinguished by their specificity, environment-friendliness and the ability to react under mild conditions can be easily subject of denaturation or inhibition. With regard to the usually high cost of purchase, the use of these enzymes could often be disadvantageous. Immobilization techniques offer an efficient solution to this problem and greatly simplify the use of enzymes in industry and research. Compared to the free forms, immobilized enzymes show greater activity, stability and allow repeated use as well as easier separation from products. This thesis contains an overview of the basic methods of immobilization - physical absorption and covalent bonds to the carrier, entrapment, encapsulation and carrier- free techniques using cross-linking. Finally, we outline possible biomedical applications as well as the use of immobilised enzymes in biosensors.
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Protein macromolecules immobilization onto polymer carriers
Šitnerová, Michaela ; Holas, Ondřej (advisor) ; Ondrejček, Pavel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of: Pharmaceutical Technology Consultant: PharmDr. Ondřej Holas, Ph.D. Student: Michaela Šitnerová Title of Thesis: Protein macromolecules immobilization onto polymer carriers Enzymes are unique biocatalysts because of their properties. They are highly specific, selective and functional even under mild reaction conditions. The method of immobilization is used to increase their operational stability, activity and possible reuse. This process allows the wide use of enzymes in industry, for example in the food industry, analytical chemistry, chemical synthesis and in the pharmaceutical industry. The aim of my thesis was immobilized enzyme acetylcholinesterase (AChE) on the surface pellets of microcrystalline cellulose (MCC). Used method was simple sorption, immobilization using glutaraldehyde, and TEMPO oxidation using MCC. Well known Ellman's method served to measure the activity of AChE. The absorbance of the solution with the immobilized AChE was measured spectrophotometrically at 412 nm.
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