Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism

The pyridine nucleotides NAD(H) and NADP(H) play major roles in the formation of by-products. To analyse how Saccharomyces cerevisiae (S. cerevisiae) metabolism during growth on glucose might be altered when intracellular NADH pool is decreased, we expressed noxE encoding a water-forming NADH oxidas...

Full description

Saved in:
Bibliographic Details
Published in:Metabolic engineering 2006-07, Vol.8 (4), p.303-314
Main Authors: Heux, Stéphanie, Cachon, Rémy, Dequin, Sylvie
Format: Article
Language:English
Subjects:
Cell Proliferation
Coenzymes - genetics
Coenzymes - metabolism
Energy Metabolism - physiology
Glucose - metabolism
Life Sciences
Multienzyme Complexes - genetics
Multienzyme Complexes - metabolism
NADH, NADPH Oxidoreductases - genetics
NADH, NADPH Oxidoreductases - metabolism
Oxidation-Reduction
Oxygen - metabolism
Protein Engineering - methods
Recombinant Proteins - metabolism
Saccharomyces cerevisiae - physiology
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
Water - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The pyridine nucleotides NAD(H) and NADP(H) play major roles in the formation of by-products. To analyse how Saccharomyces cerevisiae (S. cerevisiae) metabolism during growth on glucose might be altered when intracellular NADH pool is decreased, we expressed noxE encoding a water-forming NADH oxidase from Lactococcus lactis (L. lactis) in the S. cerevisiae strain V5. During batch fermentation under controlled microaeration conditions, expression of the NADH oxidase under the control of a yeast promoter lead to large decreases in the intracellular NADH concentration (five-fold) and NADH/NAD+ ratio (six-fold). This increased NADH consumption caused a large redistribution of metabolic fluxes. The ethanol, glycerol, succinate and hydroxyglutarate yields were significantly reduced as a result of the lower NADH availability, whereas the formation of more oxidized metabolites, acetaldehyde, acetate and acetoin was favoured. The biomass yield was low and consumption of glucose, at concentration above 10%, was impaired. The metabolic redistribution in cells expressing the NADH oxidase was a consequence of the maintenance of a redox balance and of the management of acetaldehyde formation, which accumulated at toxic levels early in the process.
ISSN:1096-7176
1096-7184
DOI:10.1016/j.ymben.2005.12.003