Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 231-239 |
| Seitenumfang | 9 |
| Fachzeitschrift | Bioresource technology |
| Jahrgang | 108 |
| Publikationsstatus | Veröffentlicht - 24 Dez. 2011 |
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.
ASJC Scopus Sachgebiete
- Chemische Verfahrenstechnik (insg.)
- Bioengineering
- Umweltwissenschaften (insg.)
- Environmental engineering
- Energie (insg.)
- Erneuerbare Energien, Nachhaltigkeit und Umwelt
- Umweltwissenschaften (insg.)
- Abfallwirtschaft und -entsorgung
Ziele für nachhaltige Entwicklung
Zitieren
- Standard
- Harvard
- Apa
- Vancouver
- BibTex
- RIS
in: Bioresource technology, Jahrgang 108, 24.12.2011, S. 231-239.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Functional expression of a valencene dioxygenase from Pleurotus sapidus in E. coli
AU - Zelena, Kateryna
AU - Krings, Ulrich
AU - Berger, Ralf G.
N1 - Funding information: Support of the work by the BMBF cluster Biokatalyse2021 (FKZ0315172B) is gratefully acknowledged.
PY - 2011/12/24
Y1 - 2011/12/24
N2 - 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.
AB - 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.
KW - Basidiomycete
KW - Chaperone
KW - Heterologous expression
KW - Nootkatone
KW - Oxygenase
UR - http://www.scopus.com/inward/record.url?scp=84857238768&partnerID=8YFLogxK
U2 - 10.1016/j.biortech.2011.12.097
DO - 10.1016/j.biortech.2011.12.097
M3 - Article
C2 - 22264428
AN - SCOPUS:84857238768
VL - 108
SP - 231
EP - 239
JO - Bioresource technology
JF - Bioresource technology
SN - 0960-8524
ER -