Enhancing active substance production of genetically transformed hairy root cultures, using fermentation

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Supervisor:
Bányai Péter
SE Biomedical Engineering MSc

During our task, we were dealing with cultivation of genetically modified Rubia tinctorum hairy root cultures using fermentation.

Rubia tinctorum L. (European madder) is a perennial plant from the Rubiaceae family. It is a source of a natural dye; it produces a variety of anthraquinone pigments in its roots and rhizomes. These substances show bactericidal, antifungal and spasmolytic activity and facilitate the loosening of kidney concrements.

Genetically modified Rubia tinctorum hairy root cultures provide a good perspective in active substance production, thanks to its high biosynthetical properties. During our experimental work, our aim is producing biomass in higher scale, to increase pharmacologically active substance production.

Within this thesis, first I review the basic theories, in connection with the topic, the Rubia tinctorum plant, the hairy root cultures, the anthraquinones, the fermentation of hairy root cultures, and different types of bioreactors, and the chromatographical methods we used, like solid phase extraction, and HPLC.

After the review of the basic theories, I demonstrate our experimental work. Later, I move on how we cultivated the Rubia tinctorum hairy root cultures in the laboratory, including the media, and media production used for these cultures. I introduce the fermentor, the fermentation process including the preparation, and also the collection and lyophilization of the biomass. On the end of the thesis, I demonstrate the results of spectrophotometric and HPLC analysis of global anthraquinones, alizarin, and purpurin concentrations. I assess our experiences, and refer to the further perspectives of our experimental work.

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