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| 001 | 877282 | ||
| 005 | 20220930130241.0 | ||
| 020 | _ | _ | |a 978-3-95806-480-5 |
| 024 | 7 | _ | |2 Handle |a 2128/25338 |
| 024 | 7 | _ | |2 URN |a urn:nbn:de:0001-2020072314 |
| 024 | 7 | _ | |2 ISSN |a 1866-1807 |
| 037 | _ | _ | |a FZJ-2020-02105 |
| 041 | _ | _ | |a English |
| 100 | 1 | _ | |0 P:(DE-Juel1)168455 |a Milke, Lars |b 0 |e Corresponding author |g male |u fzj |
| 245 | _ | _ | |a Engineering of Corynebacterium glutamicum towards increased malonyl-CoA availability for polyketide synthesis |f 2017-01-01 - 2020-05-18 |
| 260 | _ | _ | |a Jülich |b Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag |c 2020 |
| 300 | _ | _ | |a IX, 117 S. |
| 336 | 7 | _ | |2 DataCite |a Output Types/Dissertation |
| 336 | 7 | _ | |0 PUB:(DE-HGF)3 |2 PUB:(DE-HGF) |a Book |m book |
| 336 | 7 | _ | |2 ORCID |a DISSERTATION |
| 336 | 7 | _ | |2 BibTeX |a PHDTHESIS |
| 336 | 7 | _ | |0 2 |2 EndNote |a Thesis |
| 336 | 7 | _ | |0 PUB:(DE-HGF)11 |2 PUB:(DE-HGF) |a Dissertation / PhD Thesis |b phd |m phd |s 1595832391_14593 |
| 336 | 7 | _ | |2 DRIVER |a doctoralThesis |
| 490 | 0 | _ | |a Schriften des Forschungszentrums Jülich. Reihe Schlüsseltechnologien / Key Technologies |v 223 |
| 500 | _ | _ | |a Biotechnologie 1 |
| 502 | _ | _ | |a Heinrich-Heine-Universität Düsseldorf, Diss., 2020 |b Dr. |c Heinrich-Heine-Universität Düsseldorf |d 2020 |
| 520 | _ | _ | |a Polyketides are a structurally highly diverse group of natural products with interesting healthpromoting effects on humans. Despite all structural differences, polyketides are synthesized from simple CoA-activated carboxylic acid derivatives, such as acetyl-CoA or malonyl-CoA following a mechanism closely related to fatty acid biosynthesis. Unfortunately, polyketides are only synthesized in small quantities by the respective native organism. In contrast, microbial polyketide synthesis using engineered bacteria is a promising approach to get access to the desired products. Against this background, $\textit{Corynebacterium glutamicum}$ strains for the production of different plant polyketides such as stilbenes and flavonoids have been constructed recently. However, it soon became evident that the intracellular availability of malonyl-CoA is limiting the overall product formation in these strains. Hence, the main goal of this thesis was to optimize the intracellular malonyl-CoA availability in $\textit{C. glutamicum}$ by metabolic engineering. Additionally, the tailored strains should be used for establishing synthesis of biotechnological interesting polyketides. The following results were obtained: 1) Reduction of the citrate synthase activity to 5.5 % compared to the $\textit{C. glutamicum}$ wild type by exchanging the promotor of the encoding $\textit{gltA}$ gene, reduced acetyl-CoA consumption via the tricarboxylic acid cycle, which in turn improved malonyl-CoA availability. Upon transcriptional deregulation of $\textit{accBC}$ and $\textit{accD1}$ encoding the two subunits of acetyl-CoA carboxylase, malonyl-CoA synthesis from acetyl-CoA was drastically improved allowing for the synthesis of 65 mg/L (0.24 mM) naringenin und 450 mg/L (1.97 mM) resveratrol. Furthermore, improving the glucose uptake and elimination of anaplerotic pyruvate carboxylation reaction further contributed to an improved intracellular malonyl-CoA availability in the ultimately constructed strain $\textit{C. glutamicum}$ M-CoA. 2) Through episomal expression of genes encoding heterologous type III polyketide synthases from various plant species in the constructed strain $\textit{C. glutamicum}$ M-CoA, microbial synthesis of a pentaketide (noreugenin) but also phenylbutanoids (raspberry ketone, zingerone, benzylacetone) with a $\textit{ldhA}$-deficient variant could be established. The respective strains allowed for the synthesis of up to 53.3 mg/L (0.28 mM) noreugenin, 100 mg/L (0.61 mM) raspberry ketone, 70 mg/L (0.36 mM) zingerone and 10.5 mg/L (0.07 mM) benzylacetone from simple precursor molecules, respectively. 3) Hitherto, only type III polyketides can be synthesized by engineered $\textit{C. glutamicum}$ strains. In the context of this study, functional expression of a codon-optimized gene variant encoding the type I polyketide synthase 6-methylsalicylic acid synthase ChlB1 from $\textit {Streptomyces antibioticus}$ of 1,756 amino acids size was achieved. This allowed for the synthesis of up to 41 mg/L (0.27 mM) 6-methylsalicylic acid. It was found that $\textit{C. glutamicum}$ has an endogenous phosphopantetheinyltransferase activity, which can post-translationally activate ChlB1. This makes $\textit{C. glutamicum}$ a promising host for the production of other interesting type I polyketides. |
| 536 | _ | _ | |0 G:(DE-HGF)POF3-581 |a 581 - Biotechnology (POF3-581) |c POF3-581 |f POF III |x 0 |
| 856 | 4 | _ | |u https://juser.fz-juelich.de/record/877282/files/Schluesseltech_223.pdf |y OpenAccess |
| 856 | 4 | _ | |u https://juser.fz-juelich.de/record/877282/files/Schluesseltech_223.pdf?subformat=pdfa |x pdfa |y OpenAccess |
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| 914 | 1 | _ | |y 2020 |
| 915 | _ | _ | |0 StatID:(DE-HGF)0510 |2 StatID |a OpenAccess |
| 915 | _ | _ | |0 LIC:(DE-HGF)CCBY4 |2 HGFVOC |a Creative Commons Attribution CC BY 4.0 |
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