Effective synthesis of high-content fructooligosaccharides in engineered Aspergillus niger

• Published in: Microbial cell factories Vol. 23; no. 1; p. 76
• Main Authors: Wan, Xiufen, Wang, Lu, Chang, Jingjing, Zhang, Jing, Zhang, Zhiyun, Li, Kewen, Sun, Guilian, Liu, Caixia, Zhong, Yaohua
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 09.03.2024; BioMed Central; BMC
• Subjects: Analysis; Aspergillus; Aspergillus niger; b-Fructofuranosidase; Biocatalysts; Composition; Control; Enzymes; Fermentation; Fructooligosaccharides; Genetic engineering; Genomes; Glucose; Glucose oxidase; Heterologous expression; Identification and classification; Invertase; Localization; Metabolic engineering; Methods; Oligosaccharides; Peroxidase; Polyethylene glycol; Purity; Sucrose; Synthesis; Yeast; β-fructofuranosidase; China; Glucose oxidase; Peroxidase; β-fructofuranosidase; Aspergillus niger; Fructooligosaccharides; Heterologous expression
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-024-02353-w

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. 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%. 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.