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Leitlinien Unfallchirurgie
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Lectura de prueba, PDF (2,4 MB)
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The fungus Ashbya gossypii is an important industrial producer of riboflavin, i.e. vitamin B2. Here, we developed and then used a highly sophisticated set-up of parallel 13C tracer studies with labeling analysis by GC/MS, LC/MS, 1D, and 2D NMR to resolve carbon fluxes and obtain a detailed picture of the underlying metabolism in the overproducing strain A. gossypii B2 during growth and riboflavin production from a complex industrial medium using vegetable oil as carbon source. Glycine was exclusively used as carbon-two – but not carbon-one (C1) – donor of the vitamin’s pyrimidine ring due to the proven absence of a functional glycine cleavage system. Yeast extract was the main carbon source during growth, while still contributing 8 % overall carbon to riboflavin. Overall carbon flux from rapeseed oil into riboflavin equaled 80 %. Transmembrane formate flux simulations revealed that the C1-supply displayed a severe bottleneck during the initial riboflavin production, which was overcome in later phases of the cultivation by intrinsic formate accumulation. The transiently limiting C1-pool was successfully replenished by time-resolved feeding of formate or serine. This increased the intracellular precursor availability and resulted in a riboflavin titer increase of 45 %. This study is the first that successfully sheds light on carbon fluxes during the growth and riboflavin production phase by use of 13C tracers and a complementary platform of analytical techniques.
ISBN-13 (Impresion) | 9783736998810 |
ISBN-13 (E-Book) | 9783736988811 |
Idioma | Inglés |
Numero de paginas | 174 |
Laminacion de la cubierta | mate |
Edicion | 1. |
Serie | Hochschulschriften - Institut für Systembiotechnologie, Universität des Saarlandes |
Volumen | 6 |
Lugar de publicacion | Göttingen |
Lugar de la disertacion | Saarbrücken |
Fecha de publicacion | 26.11.2018 |
Clasificacion simple | Tesis doctoral |
Area |
Microbiología y biotécnica
|
Palabras claves | Ashbya gossypii, Riboflavin, Vitamin B2, 13C tracer, 13C Metabolic flux analysis, Isotope labeling, GC/MS, LC/MS, NMR, Yeast extract, Industrial process, Systems biotechnology, Industrial biotechnology, Metabolic network analysis, Bacillus subtilis, Carbon-one metabolism, Glycine metabolism, Vegetable oil, Rapeseed oil, Overproducer, Vitamin, Riboflavin biosynthesis, Purine biosynthesis, Metabolic engineering, Positional enrichment, Biotechnology, Parallel tracer experiments |