Multiple gene integration to promote erythritol production on glycerol in Yarrowia lipolytica

Biotechnol Lett. 2021 Jul;43(7):1277-1287. doi: 10.1007/s10529-021-03113-1. Epub 2021 Apr 2.

Abstract

Objective: Erythritol (1,2,3,4-butanetetrol) is a 4-carbon sugar alcohol that occurs in nature as a metabolite or storage compound. In this study, a multiple gene integration strategy was employed to enhance erythritol production in Y. lipolytica.

Results: The effects on the production of erythritol in Y. lipolytica of seven key genes involved in the erythritol synthesis pathway were evaluated individually, among which transketolase (TKL1) and transaldolase (TAL1) showed important roles in enhancing erythritol production. The combined overexpression of four genes (GUT1, TPI1, TKL1, TAL1) and disruption of the EYD1 gene (encoding erythritol dehydrogenase), resulted in produce approximately 40 g/L erythritol production from glycerol. Further enhanced erythritol synthesis was obtained by overexpressing the RKI1 gene (encoding ribose 5-phosphate isomerase) and the AMPD gene (encoding AMP deaminase), indicating for the first time that these two genes are also related to the enhancement of erythritol production in Y. lipolytica.

Conclusions: A combined gene overexpression strategy was developed to efficiently improve the production of erythritol in Y. lipolytica, suggesting a great capacity and promising potential of this non-conventional yeast in converting glycerol into erythritol.

Keywords: Erythritol; Glycerol utilization; Multiple gene integration; Ribose 5-phosphate isomerase; Yarrowia lipolytica.

MeSH terms

  • AMP Deaminase / genetics
  • Aldose-Ketose Isomerases / genetics
  • Batch Cell Culture Techniques
  • Erythritol / biosynthesis*
  • Fungal Proteins / genetics*
  • Glycerol / metabolism
  • Metabolic Engineering / methods*
  • Transaldolase / genetics
  • Transketolase / genetics
  • Yarrowia / genetics
  • Yarrowia / growth & development*
  • Yarrowia / metabolism

Substances

  • Fungal Proteins
  • Transketolase
  • Transaldolase
  • AMP Deaminase
  • Aldose-Ketose Isomerases
  • ribosephosphate isomerase
  • Glycerol
  • Erythritol