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Evaluation of different expression systems for the heterologous expression of pyranose 2-oxidase from Trametes multicolor in E. coli
KTH, School of Biotechnology (BIO).
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2010 (English)In: Microbial Cell Factories, ISSN 1475-2859, Vol. 9, 14- p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2010. Vol. 9, 14- p.
National Category
Industrial Biotechnology
URN: urn:nbn:se:kth:diva-28426DOI: 10.1186/1475-2859-9-14ISI: 000275734700001ScopusID: 2-s2.0-77950413255OAI: diva2:388386
QC 20110117Available from: 2011-01-17 Created: 2011-01-14 Last updated: 2011-01-17Bibliographically approved

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