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 (Corresponding Author), Hannu Maaheimo, Merja Penttilä, Laura Ruohonen

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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 13C-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 (≥2 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
JournalApplied Microbiology and Biotechnology
Volume85
Issue number3
DOIs
Publication statusPublished - 1 Jan 2010
MoE publication typeA1 Journal article-refereed

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Ribitol
Pentose Phosphate Pathway
Ribose
Saccharomyces cerevisiae
Glucose
Glucose-6-Phosphate Isomerase
Fructose
Transketolase
Sugar Phosphates
Glutamate Dehydrogenase
Glyceraldehyde-3-Phosphate Dehydrogenases
Growth
Bacillus subtilis
NADP
NAD
Genes
Magnetic Resonance Spectroscopy

Keywords

  • D-ribose
  • NMR
  • Pentose phosphate pathway
  • Pentose sugars
  • Ribitol
  • Saccharomyces cerevisiae
  • Sugar alcohols

Cite this

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title = "Enhancing the flux of D-glucose to the pentose phosphate pathway in Saccharomyces cerevisiae for the production of D-ribose and ribitol",
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 13C-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 (≥2 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.",
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T1 - Enhancing the flux of D-glucose to the pentose phosphate pathway in Saccharomyces cerevisiae for the production of D-ribose and ribitol

AU - Toivari, Mervi H.

AU - Maaheimo, Hannu

AU - Penttilä, Merja

AU - Ruohonen, Laura

PY - 2010/1/1

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N2 - 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 13C-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 (≥2 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.

AB - 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 13C-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 (≥2 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.

KW - D-ribose

KW - NMR

KW - Pentose phosphate pathway

KW - Pentose sugars

KW - Ribitol

KW - Saccharomyces cerevisiae

KW - Sugar alcohols

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DO - 10.1007/s00253-009-2184-4

M3 - Article

C2 - 19711072

AN - SCOPUS:74149093593

VL - 85

SP - 731

EP - 739

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 3

ER -