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Novel strategies for automated engineering of enzymatic systems: structural and functional insights to human pyruvate dehydrogenase complex
Citation Link: https://doi.org/10.15480/882.3290
Publikationstyp
Thesis
Thesis Type
Doctoral Thesis
Publikationsdatum
2021-02-15
Sprache
English
Author
Advisor
Referee
Title Granting Institution
Technische Universität Hamburg
Place of Title Granting Institution
Hamburg
Examination Date
2020-11-16
TORE-URI
Citation
Technische Universität Hamburg (2021)
This thesis comprises the development of a process strategy for the targeted exploitation of beneficial enzyme properties via rapid and automated protein production and screening. Furthermore, to provide a better understanding of the structure and functionality of the human pyruvate dehydrogenase complex (hPDC). Structurally, interactions of different protein domains within hPDC were investigated using truncated core variants. Moreover, the hPDC activity was examined by multi-parameter screening, whereat an oscillating behaviour of hPDC was determined experimentally under certain parameter conditions. Finally, an automated process chain was developed to manipulate DNA sequences, produce proteins and subsequently test for functionality within few hours.
Schlagworte
Pyruvate Dehydrogenase Complex
Process automation
Nonlinear enzyme dynamics
Protein interaction
DDC Class
570: Biowissenschaften, Biologie
600: Technik