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Gewählte Publikation:

Spadiut, O; Posch, G; Ludwig, R; Haltrich, D; Peterbauer, CK.
(2010): Evaluation of different expression systems for the heterologous expression of pyranose 2-oxidase from Trametes multicolor in E. coli
MICROB CELL FACT. 2010; 9: FullText FullText_BOKU

The heterologous production of the industrially relevant fungal enzyme pyranose 2-oxidase in the prokaryotic host E. coli was investigated using 3 different expression systems, i.e. the well-studied T7 RNA polymerase based pET21d(+), the L-arabinose inducible pBAD and the pCOLD system. Preliminary experiments were done in shaking flasks at 25 degrees C and optimized induction conditions to compare the productivity levels of the different expression systems. The pET21d(+) and the pCOLD system gave 29 U/L.h and 14 U/L.h of active pyranose 2-oxidase, respectively, whereas the pBAD system only produced 6 U/L.h. Process conditions for batch fermentations were optimized for the pET21d(+) and the pCOLD systems in order to reduce the formation of inactive inclusion bodies. The highest productivity rate with the pET21d(+) expression system in batch fermentations was determined at 25 C with 32 U/L.h. The pCOLD system showed the highest productivity rate (19 U/L.h) at 25 degrees C and induction from the start of the cultivation. Using the pCOLD system in a fed batch fermentation at 25 degrees C with a specific growth rate of mu = 0.15 h(-1) resulted in the highest productivity rate of active pyranose oxidase with 206 U/L.h.
Autor/innen der BOKU Wien:
Haltrich Dietmar
Ludwig Roland
Peterbauer Clemens Karl
Posch Gerald
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Carbohydrate Dehydrogenases/genetics*;Carbohydrate Dehydrogenases/metabolism;Culture Techniques;Escherichia coli/genetics*;Escherichia coli/growth & development;Escherichia coli/metabolism;Fungal Proteins/genetics*;Fungal Proteins/metabolism;Gene Expression*;Genetic Engineering/methods*;Genetic Vectors/genetics;Genetic Vectors/metabolism;Trametes/enzymology*;

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