TY - THES AU - Unthan, Simon TI - Robot-assisted phenotyping of genome-reduced Corynebacterium glutamicum strain libraries to draft a chassis organism VL - 132 PB - RWTH Aachen University VL - Dissertation CY - Jülich M1 - RWTH-2016-07483 T2 - Schriften des Forschungszentrums Jülich. Reihe Schlüsseltechnologien / Key Technologies SP - 1 Online-Ressource (122 Seiten) : Illustrationen, Diagramme PY - 2016 N1 - Druckausgabe: 2016. - Onlineausgabe: 2016. - Auch veröffentlicht auf dem Publikationsserver der RWTH Aachen University N1 - Dissertation, RWTH Aachen University, 2015 AB - In this work, concepts were developed and applied to guide the construction of a Corynebacterium glutamicum chassis organism for synthetic biology approaches. The aim was to delete irrelevant genes from the wild type strain in order to obtain a chassis growing on defined CGXII medium with D-glucose with unaltered biological fitness, which was defined by the maximum specific growth rate and biomass yield.Initially, workflows were developed on a robotic Mini Pilot Plant (MPP), for example, to harvest cell-free cultivation supernatants from BioLector cultivations in response to individually defined triggers. Subsequently, assays for amino acids and D-glucose were established in 384-well plate scale in order to quantify these metabolites in cell-free culture supernatants in fully automated workflows [1].During initial reference experiments, protocatechuic acid was identified as a hidden co-substrate in the well-known defined CGXII medium. The additional TCA feed via acetyl-CoA and succinyl-CoA, which are derived from protocatechuic acid, elevates the growth rate by about 50 LB - PUB:(DE-HGF)11 ; PUB:(DE-HGF)3 UR - https://publications.rwth-aachen.de/record/670366 ER -