Bionanotechnology is a broad field that is receiving increasing attention in society for many economic, environmental, health and social reasons. An important part of bionanotechnology is the development of new materials that could be used for various purposes. With synthetic biology, which introduces engineering approaches to biotechnology, we can design and prepare completely new bionanomaterials based on natural protein building blocks and those that do not exist in nature. In order to prepare bionanostructures that self-assemble into the planned structure, it is necessary to first carefully design the building blocks for the planned nanostructure or material, and then to acquire and characterize these basic building blocks, which is the focus of this master's thesis.
The 34cald56, the 34cald56 version with fluorescence protein and the 3APH4 polypeptide building blocks were prepared for the purpose of constructing two designed two-dimensional protein lattices having different geometries and self-assembling from one polypeptide building block. All three proteins were successfully produced in bacteria E. coli, isolated and characterized with biochemical methods.
We have confirmed that the secondary structure of the 34cald56 and the 3APH4 polypeptide building blocks is in accordance with the designed. From the data of the circular dichroism measurements, we concluded that the temperature stability of the protein 34cald56 is higher than the temperature stability of the 3APH4. Measurements of the dynamic light scattering have revealed that the folded polypeptides 34cald56 and 3APH4 in solution self-assemble into dimers and larger nanostructures that become insoluble and precipitate out of solution, but can also be assembled into soluble nanostructures, which in the case of 34cald56 are large around 120 nm, and in the case of 3APH4 about 60 nm.
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