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- Title
Production of d-glucaric acid with phosphoglucose isomerase-deficient Saccharomyces cerevisiae.
- Authors
Toivari, Mervi; Vehkomäki, Maija-Leena; Ruohonen, Laura; Penttilä, Merja; Wiebe, Marilyn G.
- Abstract
d-Glucaric acid is a potential biobased platform chemical. Previously mainly Escherichia coli, but also the yeast Saccharomyces cerevisiae, and Pichia pastoris, have been engineered for conversion of d-glucose to d-glucaric acid via myo-inositol. One reason for low yields from the yeast strains is the strong flux towards glycolysis. Thus, to decrease the flux of d-glucose to biomass, and to increase d-glucaric acid yield, the four step d-glucaric acid pathway was introduced into a phosphoglucose isomerase deficient (Pgi1p-deficient) Saccharomyces cerevisiae strain. High d-glucose concentrations are toxic to the Pgi1p-deficient strains, so various feeding strategies and use of polymeric substrates were studied. Uniformly labelled 13C-glucose confirmed conversion of d-glucose to d-glucaric acid. In batch bioreactor cultures with pulsed d-fructose and ethanol provision 1.3 g d-glucaric acid L−1 was produced. The d-glucaric acid titer (0.71 g d-glucaric acid L−1) was lower in nitrogen limited conditions, but the yield, 0.23 g d-glucaric acid [g d-glucose consumed]−1, was among the highest that has so far been reported from yeast. Accumulation of myo-inositol indicated that myo-inositol oxygenase activity was limiting, and that there would be potential to even higher yield. The Pgi1p-deficiency in S. cerevisiae provides an approach that in combination with other reported modifications and bioprocess strategies would promote the development of high yield d-glucaric acid yeast strains.
- Subjects
SACCHAROMYCES cerevisiae; PICHIA pastoris; ACIDS; ESCHERICHIA coli; ISOMERASES
- Publication
Biotechnology Letters, 2024, Vol 46, Issue 1, p69
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-023-03443-2