@phdthesis{Moratti2020, author = {Moratti, Fabio Giulio}, title = {Structural analysis of DYW proteins and identification of the mitochondrial DNA-binding proteome of Arabidopsis thaliana}, school = {Universit{\"a}t Potsdam}, pages = {164}, year = {2020}, language = {en} } @phdthesis{Wenk2020, author = {Wenk, Sebastian}, title = {Engineering formatotrophic growth in Escherichia coli}, school = {Universit{\"a}t Potsdam}, pages = {V, 107}, year = {2020}, abstract = {To meet the demands of a growing world population while reducing carbon dioxide (CO2) emissions, it is necessary to capture CO2 and convert it into value-added compounds. In recent years, metabolic engineering of microbes has gained strong momentum as a strategy for the production of valuable chemicals. As common microbial feedstocks like glucose directly compete with human consumption, the one carbon (C1) compound formate was suggested as an alternative feedstock. Formate can be easily produced by various means including electrochemical reduction of CO2 and could serve as a feedstock for microbial production, hence presenting a novel entry point for CO2 to the biosphere and a storage option for excess electricity. Compared to the gaseous molecule CO2, formate is a highly soluble compound that can be easily handled and stored. It can serve as a carbon and energy source for natural formatotrophs, but these microbes are difficult to cultivate and engineer. In this work, I present the results of several projects that aim to establish efficient formatotrophic growth of E. coli - which cannot naturally grow on formate - via synthetic formate assimilation pathways. In the first study, I establish a workflow for growth-coupled metabolic engineering of E. coli. I demonstrate this approach by presenting an engineering scheme for the PFL-threonine cycle, a synthetic pathway for anaerobic formate assimilation in E. coli. The described methods are intended to create a standardized toolbox for engineers that aim to establish novel metabolic routes in E. coli and related organisms. The second chapter presents a study on the catalytic efficiency of C1-oxidizing enzymes in vivo. As formatotrophic growth requires generation of both energy and biomass from formate, the engineered E. coli strains need to be equipped with a highly efficient formate dehydrogenase, which provides reduction equivalents and ATP for formate assimilation. I engineered a strain that cannot generate reducing power and energy for cellular growth, when fed on acetate. Under this condition, the strain depends on the introduction of an enzymatic system for NADH regeneration, which could further produce ATP via oxidative phosphorylation. I show that the strain presents a valuable testing platform for C1-oxidizing enzymes by testing different NAD-dependent formate and methanol dehydrogenases in the energy auxotroph strain. Using this platform, several candidate enzymes with high in vivo activity, were identified and characterized as potential energy-generating systems for synthetic formatotrophic or methylotrophic growth in E. coli.   In the third chapter, I present the establishment of the serine threonine cycle (STC) - a synthetic formate assimilation pathway - in E. coli. In this pathway, formate is assimilated via formate tetrahydrofolate ligase (FtfL) from Methylobacterium extorquens (M. extorquens). The carbon from formate is attached to glycine to produce serine, which is converted into pyruvate entering central metabolism. Via the natural threonine synthesis and cleavage route, glycine is regenerated and acetyl-CoA is produced as the pathway product. I engineered several selection strains that depend on different STC modules for growth and determined key enzymes that enable high flux through threonine synthesis and cleavage. I could show that expression of an auxiliary formate dehydrogenase was required to achieve growth via threonine synthesis and cleavage on pyruvate. By overexpressing most of the pathway enzymes from the genome, and applying adaptive laboratory evolution, growth on glycine and formate was achieved, indicating the activity of the complete cycle. The fourth chapter shows the establishment of the reductive glycine pathway (rGP) - a short, linear formate assimilation route - in E. coli. As in the STC, formate is assimilated via M. extorquens FtfL. The C1 from formate is condensed with CO2 via the reverse reaction of the glycine cleavage system to produce glycine. Another carbon from formate is attached to glycine to form serine, which is assimilated into central metabolism via pyruvate. The engineered E. coli strain, expressing most of the pathway genes from the genome, can grow via the rGP with formate or methanol as a sole carbon and energy source.}, language = {en} } @phdthesis{Kubis2020, author = {Kubis, Armin}, title = {Synthetic carbon neutral photorespiration bypasses}, school = {Universit{\"a}t Potsdam}, pages = {68}, year = {2020}, abstract = {With populations growing worldwide and climate change threatening food production there is an urgent need to find ways to ensure food security. Increasing carbon fixation rate in plants is a promising approach to boost crop yields. The carbon-fixing enzyme Rubisco catalyzes, beside the carboxylation reaction, also an oxygenation reaction that generates glycolate-2P, which needs to be recycled via a metabolic route termed photorespiration. Photorespiration dissipates energy and most importantly releases previously fixed CO2, thus significantly lowering carbon fixation rate and yield. Engineering plants to omit photorespiratory CO2 release is the goal of the FutureAgriculture consortium and this thesis is part of this collaboration. The consortium aims to establish alternative glycolate-2P recycling routes that do not release CO2. Ultimately, they are expected to increase carbon fixation rates and crop yields. Natural and novel reactions, which require enzyme engineering, were considered in the pathway design process. Here I describe the engineering of two pathways, the arabinose-5P and the erythrulose shunt. They were designed to recycle glycolate-2P via glycolaldehyde into a sugar phosphate and thereby reassimilate glycolate-2P to the Calvin cycle. I used Escherichia coli gene deletion strains to validate and characterize the activity of both synthetic shunts. The strains' auxotrophies can be alleviated by the activity of the synthetic route, thus providing a direct way to select for pathway activity. I introduced all pathway components to these dedicated selection strains and discovered inhibitions, limitations and metabolic cross talk interfering with pathway activity. After resolving these issues, I was able to show the in vivo activity of all pathway components and combine them into functional modules.. Specifically, I demonstrate the activity of a new-to-nature module of glycolate reduction to glycolaldehyde. Also, I successfully show a new glycolaldehyde assimilation route via arabinose-5P to ribulose-5P. In addition, all necessary enzymes for glycolaldehyde assimilation via L-erythrulose were shown to be active and an L-threitol assimilation route via L-erythrulose was established in E. coli. On their own, these findings demonstrate the power of using an easily engineerable microbe to test novel pathways; combined, they will form the basis for implementing photorespiration bypasses in plants.}, language = {en} } @phdthesis{Kožul2020, author = {Kožul, Danijela}, title = {Systematic identification of loci determining chloroplast and nuclear genome incompatibility in the evening primrose (Oenothera)}, school = {Universit{\"a}t Potsdam}, pages = {126}, year = {2020}, language = {en} } @phdthesis{Kehr1998, author = {Kehr, Julia}, title = {Mikroanalyse einzelner Zellen und Kompartimente transgener Pflanzen mittels biophysikalischer Methoden}, address = {Potsdam}, pages = {VII, 102 S. : graph. Darst.}, year = {1998}, language = {de} } @phdthesis{FuentesTaladriz2015, author = {Fuentes Taladriz, Paulina Andrea}, title = {High-level production of the antimalarial drug precursor artemisinic acid in plastids and in vivo visualization of plastid-to-nucleus gene transfer}, school = {Universit{\"a}t Potsdam}, pages = {148}, year = {2015}, language = {en} } @phdthesis{Bissinger2003, author = {Bissinger, Vera}, title = {Factors determining growth and vertical distribution of planktonic algae in extremely acidic mining lakes (pH 2.7)}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-0000695}, school = {Universit{\"a}t Potsdam}, year = {2003}, abstract = {Die vorliegende Dissertation besch{\"a}ftigt sich mit den Faktoren, die das Wachstum und die Vertikalverteilung von Planktonalgen in extrem sauren Tagebaurestseen (TBS; pH 2-3) beeinflussen. Im exemplarisch untersuchten TBS 111 (pH 2.7; Lausitzer Revier) dominiert die Goldalge Ochromonas sp. in oberen und die Gr{\"u}nalge Chlamydomonas sp. in tieferen Wasserschichten, wobei letztere ein ausgepr{\"a}gtes Tiefenchlorophyll-Maximum (DCM) ausbildet. Es wurde ein deutlicher Einfluss von Limitation durch anorganischen Kohlenstoff (IC) auf das phototrophe Wachstum von Chlamydomonas sp. in oberen Wasserschichten nachgewiesen, die mit zunehmender Tiefe von Lichtlimitation abgel{\"o}st wird. Im Vergleich mit Arbeiten aus neutralen Seen zeigte Chlamydomonas sp. erniedrigte maximale Wachstumsraten, einen gesteigerten Kompensationspunkt und erh{\"o}hte Dunkelrespirationsraten, was auf gesteigerte metabolische Kosten unter den extremen physikalisch-chemischen Bedingungen hinweist. Die Photosyntheseleistungen von Chlamydomonas sp. waren in Starklicht-adaptierten Zellen durch IC-Limitation deutlich verringert. Außerdem ergaben die ermittelten minimalen Zellquoten f{\"u}r Phosphor (P) einen erh{\"o}hten P-Bedarf unter IC-Limitation. Anschließend konnte gezeigt werden, dass Chlamydomonas sp. ein mixotropher Organismus ist, der seine Wachstumsraten {\"u}ber die osmotrophe Aufnahme gel{\"o}sten organischen Kohlenstoffs (DOC) erh{\"o}hen kann. Dadurch ist dieser Organismus f{\"a}hig, in tieferen, Licht-limitierten Wasserschichten zu {\"u}berleben, die einen h{\"o}heren DOC-Gehalt aufweisen. Da die Vertikalverteilung der Algen im TBS 111 jedoch weder durch IC-Limitation, P-Verf{\"u}gbarkeit noch die in situ DOC-Konzentrationen abschließend erkl{\"a}rt werden konnte (bottom-up Kontrolle), wurde eine neue Theorie zur Entstehung der Vertikalverteilung gepr{\"u}ft. Grazing der phagotrophen und phototrophen Alge Ochromonas sp. auf der phototrophen Alge Chlamydomonas sp. erwies sich als herausragender Faktor, der {\"u}ber top-down Kontrolle die Abundanz der Beute in h{\"o}heren Wasserschichten beeinflussen kann. Gemeinsam mit der Tatsache, dass Chlamydomonas sp. DOC zur Wachstumssteigerung verwendet, f{\"u}hrt dies zu einer Akkumulation von Chlamydomonas sp. in der Tiefe, ausgepr{\"a}gt als DCM. Daher erscheint grazing als der Hauptfaktor, der die beobachtete Vertikalschichtung der Algen im TBS 111 hervorruft. Die erzielten Ergebnisse liefern grundlegende Informationen, um die Auswirkungen von Strategien zur Neutralisierung der TBS auf das Nahrungsnetz absch{\"a}tzen zu k{\"o}nnen.}, language = {en} } @phdthesis{Artins2023, author = {Artins, Anthony}, title = {Crosstalk between Target Of Rapamycin (TOR) and sugar signaling in Arabidopsis thaliana}, school = {Universit{\"a}t Potsdam}, pages = {125}, year = {2023}, language = {en} } @phdthesis{Ting2021, author = {Ting, Michael Kien Yin}, title = {Circadian-regulated dynamics of translation in Arabidopsis thaliana}, school = {Universit{\"a}t Potsdam}, pages = {130}, year = {2021}, language = {en} } @phdthesis{Brunacci2021, author = {Brunacci, Nadia}, title = {Oligodepsipeptides as matrix for drug delivery systems and submicron particulate carriers}, school = {Universit{\"a}t Potsdam}, year = {2021}, language = {en} }