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Testing of biological effect of glucans on cell lines in vitro
Šimková, Eva ; Holub, Jiří (referee) ; Němcová, Andrea (advisor)
The aim of this bachelor thesis is isolation and testing of -glucans from selected strains of yeast. In the theoretical part, characterisation of -glucans, microbial production of -glucans by yeast, determination of polysaccharides, exopolysaccharides and in vitro testing are described. Yeast have a cell wall made up of -glucans and -glucans are known for their healing properties. In the experimental part of this work deals with characterization of selected biomass yeasts in different forms and comparison of their effects and the amount of -glucans in each form. Stains of Metschnikowia pulcherrima, Cystofilobasidium infirmominiatum and Rhodotorula kratochvilovae have been selected for this thesis. Subsequently, the thesis deals with the isolation and testing of isolated -glucans from a selected lyophilised strain of yeast Rhodotorula kratochvilovae. The selected strain was isolated in 2 ways. The first method of isolation that was more friendly to conformation of -glucans due to the effect of ultrasonic homogenization of cells. The second method of isolation was less friendly to -glucans, using acid and base to homogenise them. Analyses of both extraction methods confirmed the presence of isolated -glucans, however it was found that the recovery of both methods was relatively low and requires further optimisation of the procedure. Overall, the better extraction procedure was the second method. A sample of this extract was further tested on the HaCaT cell line. The results of the pilot tests confirmed the cytotoxicity of the extract obtained by us.
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Testing of biological effect of glucans on cell lines in vitro
Šimková, Eva ; Holub, Jiří (referee) ; Němcová, Andrea (advisor)
The aim of this bachelor thesis is isolation and testing of -glucans from selected strains of yeast. In the theoretical part, characterisation of -glucans, microbial production of -glucans by yeast, determination of polysaccharides, exopolysaccharides and in vitro testing are described. Yeast have a cell wall made up of -glucans and -glucans are known for their healing properties. In the experimental part of this work deals with characterization of selected biomass yeasts in different forms and comparison of their effects and the amount of -glucans in each form. Stains of Metschnikowia pulcherrima, Cystofilobasidium infirmominiatum and Rhodotorula kratochvilovae have been selected for this thesis. Subsequently, the thesis deals with the isolation and testing of isolated -glucans from a selected lyophilised strain of yeast Rhodotorula kratochvilovae. The selected strain was isolated in 2 ways. The first method of isolation that was more friendly to conformation of -glucans due to the effect of ultrasonic homogenization of cells. The second method of isolation was less friendly to -glucans, using acid and base to homogenise them. Analyses of both extraction methods confirmed the presence of isolated -glucans, however it was found that the recovery of both methods was relatively low and requires further optimisation of the procedure. Overall, the better extraction procedure was the second method. A sample of this extract was further tested on the HaCaT cell line. The results of the pilot tests confirmed the cytotoxicity of the extract obtained by us.
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