Enhancing the flux of D-glucose to the pentose phosphate pathway in Saccharomyces cerevisiae for the production of D-ribose and ribitol

Mervi H. Toivari*, Hannu Maaheimo, Merja Penttila, Laura Ruohonen

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Phosphoglucose isomerase-deficient (pgi1) strains of Saccharomyces cerevisiae were studied for the production of D-ribose and ribitol from D-glucose via the intermediates of the pentose phosphate pathway. Overexpression of the genes coding for NAD(+)-specific glutamate dehydrogenase (GDH2) of S. cerevisiae or NADPH-utilising glyceraldehyde-3-phosphate dehydrogenase (gapB) of Bacillus subtilis enabled growth of the pgi1 mutant strains on D-glucose. Overexpression of the gene encoding sugar phosphate phosphatase (DOG1) of S. cerevisiae was needed for the production of D-ribose and ribitol; however, it reduced the growth of the pgi1 strains expressing GDH2 or gapB in the presence of higher D-glucose concentrations. The CEN.PK2-1D laboratory strain expressing both gapB and DOG1 produced approximately 0.4 g l(-1) of D-ribose and ribitol when grown on 20 g l(-1) (w/v) D-fructose with 4 g l(-1) (w/v) D-glucose. Nuclear magnetic resonance measurements of the cells grown with (13)C-labelled D-glucose showed that about 60% of the D-ribose produced was derived from D-glucose. Strains deficient in both phosphoglucose isomerase and transketolase activities, and expressing DOG1 and GDH2 tolerated only low D-glucose concentrations (a parts per thousand currency sign2 g l(-1) (w/v)), but produced 1 g l(-1) (w/v) D-ribose and ribitol when grown on 20 g l(-1) (w/v) D-fructose with 2 g l(-1) (w/v) D-glucose.

Original languageEnglish
Pages (from-to)731-739
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume85
Issue number3
DOIs
Publication statusPublished - Jan 2010
MoE publication typeA1 Journal article-refereed

Keywords

  • Sugar alcohols
  • Pentose sugars
  • Saccharomyces cerevisiae
  • Pentose phosphate pathway
  • D-ribose
  • Ribitol
  • NMR
  • KLUYVEROMYCES-LACTIS
  • BACILLUS-SUBTILIS
  • ESCHERICHIA-COLI
  • ISOMERASE MUTANT
  • AMINO-ACIDS
  • STRAINS
  • YEAST
  • IDENTIFICATION
  • METABOLISM
  • VECTORS

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