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- Source :
- Microbial Cell Factories.
-
Abstract
- meso-Galactaric acid is a dicarboxylic acid that can be produced by the oxidation of d-galacturonic acid, the main constituent of pectin. Mould strains can be engineered to perform this oxidation by expressing the bacterial enzyme uronate dehydrogenase. In addition, the endogenous pathway for d-galacturonic acid catabolism has to be inactivated. The filamentous fungus Aspergillus niger would be a suitable strain for galactaric acid production since it is efficient in pectin hydrolysis, however, it is catabolizing the resulting galactaric acid via an unknown catabolic pathway. In this study, a transcriptomics approach was used to identify genes involved in galactaric acid catabolism. Several genes were deleted using CRISPR/Cas9 together with in vitro synthesized sgRNA. As a result, galactaric acid catabolism was disrupted. An engineered A. niger strain combining the disrupted galactaric and d-galacturonic acid catabolism with an expression of a heterologous uronate dehydrogenase produced galactaric acid from d-galacturonic acid. The resulting strain was also converting pectin-rich biomass to galactaric acid in a consolidated bioprocess. In the present study, we demonstrated the use of CRISPR/Cas9 mediated gene deletion technology in A. niger in an metabolic engineering application. As a result, a strain for the efficient production of galactaric acid from d-galacturonic acid was generated. The present study highlights the usefulness of CRISPR/Cas9 technology in the metabolic engineering of filamentous fungi.
- Subjects :
- 0106 biological sciences
0301 basic medicine
2. Zero hunger
chemistry.chemical_classification
biology
Catabolism
Aspergillus niger
Bioengineering
Mucic acid
biology.organism_classification
01 natural sciences
Applied Microbiology and Biotechnology
Sugar acids
Microbiology
Metabolic engineering
03 medical and health sciences
chemistry.chemical_compound
030104 developmental biology
Dicarboxylic acid
Biochemistry
chemistry
010608 biotechnology
Uronate dehydrogenase
D-Galacturonic acid
Biotechnology
Subjects
Details
- ISSN :
- 14752859
- Database :
- OpenAIRE
- Journal :
- Microbial Cell Factories
- Accession number :
- edsair.doi...........f7eb295e74f5a2379f129400e39a41b1