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https://doi.org/10.15480/882.3290
Title: | Novel strategies for automated engineering of enzymatic systems: structural and functional insights to human pyruvate dehydrogenase complex | Language: | English | Authors: | Ilhan, Sibel | Keywords: | Pyruvate Dehydrogenase Complex;Process automation;Nonlinear enzyme dynamics;Protein interaction | Issue Date: | 15-Feb-2021 | Examination Date: | 16-Nov-2020 | Abstract (german): | Diese Arbeit umfasst die Entwicklung einer Prozessstrategie für die schnelle und automatisierte Proteinherstellung und deren anschließendes Screening zur gezielten Nutzung vorteilhafter Enzymeigenschaften. Weiterhin soll ein besseres Verständnis über Struktur und Funktionalität des humanen Pyruvatdehydrogenase Komplexes (hPDC) erlangt werden. Strukturell wurden Wechselwirkungen verschiedener Proteindomänen in hPDC mittels trunkierter Kernvarianten erforscht. Darüber hinaus wurde die hPDC-Aktivität durch ein Mehrparameter-Screening untersucht, wobei diese unter bestimmten Parameterbedingungen ein oszillierendes Verhalten aufzeigte. Schlussendlich wurde eine automatisierte Prozesskette entwickelt, welche innerhalb weniger Stunden DNA Sequenzen manipulieren, Proteine herstellen und auf Funktionalität testen kann. |
Abstract (english): | 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. |
URI: | http://hdl.handle.net/11420/8790 | DOI: | 10.15480/882.3290 | Institute: | Bioprozess- und Biosystemtechnik V-1 | Document Type: | Thesis | Thesis Type: | Doctoral Thesis | Advisor: | Zeng, An-Ping ![]() |
Referee: | Johannsen, Monika ![]() |
More Funding information: | BMBF | Project: | Biotechnologie 2020+ | License: | ![]() |
Appears in Collections: | Publications with fulltext |
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Dissertation_S._Ilhan.pdf | Novel strategies for automated engineering of enzymatic systems: Structural and functional insights to human pyruvate dehydrogenase complex --- Dissertation S. Ilhan | 8,31 MB | Adobe PDF | View/Open![]() |
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