Effective synthesis of high-content fructooligosaccharides in engineered Aspergillus niger

Microb Cell Fact. 2024 Mar 9;23(1):76. doi: 10.1186/s12934-024-02353-w.

Abstract

Background: Aspergillus niger ATCC 20611 is an industrially important fructooligosaccharides (FOS) producer since it produces the β-fructofuranosidase with superior transglycosylation activity, which is responsible for the conversion of sucrose to FOS accompanied by the by-product (glucose) generation. This study aims to consume glucose to enhance the content of FOS by heterologously expressing glucose oxidase and peroxidase in engineered A. niger.

Results: Glucose oxidase was successfully expressed and co-localized with β-fructofuranosidase in mycelia. These mycelia were applied to synthesis of FOS, which possessed an increased purity of 60.63% from 52.07%. Furthermore, peroxidase was expressed in A. niger and reached 7.70 U/g, which could remove the potential inhibitor of glucose oxidase to facilitate the FOS synthesis. Finally, the glucose oxidase-expressing strain and the peroxidase-expressing strain were jointly used to synthesize FOS, which content achieved 71.00%.

Conclusions: This strategy allows for obtaining high-content FOS by the multiple enzymes expressed in the industrial fungus, avoiding additional purification processes used in the production of oligosaccharides. This study not only facilitated the high-purity FOS synthesis, but also demonstrated the potential of A. niger ATCC 20611 as an enzyme-producing cell factory.

Keywords: Aspergillus niger; Fructooligosaccharides; Glucose oxidase; Heterologous expression; Peroxidase; β-fructofuranosidase.

MeSH terms

  • Aspergillus niger* / genetics
  • Aspergillus*
  • Glucose
  • Glucose Oxidase / genetics
  • Oligosaccharides
  • Peroxidases
  • beta-Fructofuranosidase*

Substances

  • fructooligosaccharide
  • beta-Fructofuranosidase
  • Glucose Oxidase
  • Oligosaccharides
  • Peroxidases
  • Glucose

Supplementary concepts

  • Aspergillus brasiliensis