Combination of Three Methods to Reduce Glucose Metabolic Rate for Improving N-Acetylglucosamine Production in Saccharomyces cerevisiae

Sang Woo Lee, Bo Young Lee, Min-Kyu Oh

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Previously, the production of N-acetylglucosamine (GlcNAc) in Saccharomyces cerevisiae was improved by deletion of the genes encoding phosphofructokinase 2 (PFK-2) isoforms, which reduced the glycolytic flux by eliminating the pathway to produce fructose-2,6-bisphosphate, an allosteric activator of phosphofructokinase 1 (PFK-1). We further examined the effects of an additional reduction in glucose metabolic rate on N-acetylglucosamine production. Glucose uptake rate was lowered by expressing a gene encoding truncated glucose-sensing regulator (MTH1-ΔT). In addition, catalytically dead Cas9 (dCas9) was introduced in order to down-regulate the expression levels of PFK-1 and pyruvate kinase-1 (Pyk1). Finally, the three strategies were introduced into S. cerevisiae strains in a combinatorial way; the strain containing all three modules resulted in the highest N-acetylglucosamine production yield. The results showed that the three modules cooperatively reduced the glucose metabolism and improved N-acetylglucosamine production up to 3.0 g/L in shake flask cultivation.

Original languageEnglish
Pages (from-to)13191-13198
Number of pages8
JournalJournal of Agricultural and Food Chemistry
Volume66
Issue number50
DOIs
Publication statusPublished - 2018 Dec 19

Keywords

  • dCas9
  • glucose metabolism
  • N-acetylglucosamine
  • Saccharomyces cerevisiae
  • truncated MTH1

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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