Functional expression of a valencene dioxygenase from Pleurotus sapidus in E. coli
- authored by
- Kateryna Zelena, Ulrich Krings, Ralf G. Berger
- Abstract
Valencene dioxygenase (ValOx) from the edible basidiomycete Pleurotus sapidus converted the sesquiterpene (+)-valencene to the valuable grapefruit flavour (+)-nootkatone and to nootkatols through intermediate hydroperoxides. Expression of the enzyme was carried out in the cytosol and periplasm of Escherichia coli. The heterologous production led to high yields of inclusion bodies. The poor yield of soluble recombinant protein was improved by various strategies including cold shock expression, chaperone co-expression, and employment of mutant E. coli strains. Up to 60. mg of the biologically active, soluble ValOx was produced by cold shock under control of the cspA promoter at 8 °C in the BL21(DE3)Star strain and co-expression of the E. coli trigger factor. The recombinant enzyme, purified using the N-terminal His tag, showed the catalytic properties of the wild-type enzyme, as was confirmed by the LC-MS analysis of hydroperoxide intermediates and GC-MS analysis of the volatile products.
- Organisation(s)
-
Institute of Food Chemistry
- Type
- Article
- Journal
- Bioresource technology
- Volume
- 108
- Pages
- 231-239
- No. of pages
- 9
- ISSN
- 0960-8524
- Publication date
- 24.12.2011
- Publication status
- Published
- Peer reviewed
- Yes
- ASJC Scopus subject areas
- Bioengineering, Environmental Engineering, Renewable Energy, Sustainability and the Environment, Waste Management and Disposal
- Sustainable Development Goals
- SDG 7 - Affordable and Clean Energy
- Electronic version(s)
-
https://doi.org/10.1016/j.biortech.2011.12.097 (Access:
Unknown)