Selective oxidation of UDP-glucose to UDP-glucuronic acid using permeabilized Schizosaccharomyces pombe expressing human UDP-glucose 6-dehydrogenase

Biotechnol Lett. 2016 Mar;38(3):477-81. doi: 10.1007/s10529-015-1995-x. Epub 2015 Nov 18.

Abstract

Objectives: To use permeabilized cells of the fission yeast, Schizosaccharomyces pombe, that expresses human UDP-glucose 6-dehydrogenase (UGDH, EC 1.1.1.22), for the production of UDP-glucuronic acid from UDP-glucose.

Results: In cell extracts no activity was detected. Therefore, cells were permeabilized with 0.3 % (v/v) Triton X-100. After washing away all low molecular weight metabolites, the permeabilized cells were directly used as whole cell biocatalyst. Substrates were 5 mM UDP-glucose and 10 mM NAD(+). Divalent cations were not added to the reaction medium as they promoted UDP-glucose hydrolysis. With this reaction system 5 mM UDP-glucose were converted into 5 mM UDP-glucuronic acid within 3 h.

Conclusions: Recombinant permeabilized cells of S. pombe can be used to synthesize UDP-glucuronic acid with 100 % yield and selectivity.

Keywords: Biocatalysis; Permeabilized cells; Schizosaccharomyces pombe; Selective oxidation; UDP-glucose-6-dehydrogenase; UDP-glucuronate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Detergents / metabolism
  • Glucosephosphate Dehydrogenase / genetics
  • Glucosephosphate Dehydrogenase / metabolism*
  • Humans
  • Octoxynol / metabolism
  • Oxidation-Reduction
  • Permeability / drug effects
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Schizosaccharomyces / drug effects
  • Schizosaccharomyces / metabolism*
  • Uridine Diphosphate Glucose / metabolism*
  • Uridine Diphosphate Glucuronic Acid / metabolism*

Substances

  • Detergents
  • Recombinant Proteins
  • Uridine Diphosphate Glucuronic Acid
  • Octoxynol
  • Glucosephosphate Dehydrogenase
  • Uridine Diphosphate Glucose