Genetic engineering of Synechocystis PCC6803 for the photoautotrophic production of the sweetener erythritol

Microb Cell Fact. 2016 Apr 8:15:60. doi: 10.1186/s12934-016-0458-y.

Abstract

Background: Erythritol is a polyol that is used in the food and beverage industry. Due to its non-caloric and non-cariogenic properties, the popularity of this sweetener is increasing. Large scale production of erythritol is currently based on conversion of glucose by selected fungi. In this study, we describe a biotechnological process to produce erythritol from light and CO2, using engineered Synechocystis sp. PCC6803.

Methods: By functionally expressing codon-optimized genes encoding the erythrose-4-phosphate phosphatase TM1254 and the erythrose reductase Gcy1p, or GLD1, this cyanobacterium can directly convert the Calvin cycle intermediate erythrose-4-phosphate into erythritol via a two-step process and release the polyol sugar in the extracellular medium. Further modifications targeted enzyme expression and pathway intermediates.

Conclusions: After several optimization steps, the best strain, SEP024, produced up to 2.1 mM (256 mg/l) erythritol, excreted in the medium.

Keywords: Calvin cycle; Cyanobacteria; Erythritol; Erythrose; Erythrose-4-phosphate; Metabolic engineering; Sweetener.

MeSH terms

  • Aldehyde Reductase / genetics
  • Aldehyde Reductase / metabolism
  • Autotrophic Processes
  • Erythritol / biosynthesis*
  • Escherichia coli
  • Genetic Engineering / methods*
  • Light
  • Organisms, Genetically Modified
  • Photosynthesis / genetics
  • Sugar Phosphates / metabolism
  • Sweetening Agents / metabolism
  • Synechocystis / genetics*
  • Synechocystis / growth & development*
  • Synechocystis / metabolism*

Substances

  • Sugar Phosphates
  • Sweetening Agents
  • erythrose 4-phosphate
  • Aldehyde Reductase
  • Erythritol