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)