Class
Article
College
College of Engineering
Department
Biological Engineering Department
Faculty Mentor
Jixun Zhan
Presentation Type
Poster Presentation
Abstract
Flavonoids are an important class of plant secondary metabolites with a variety of biological functions in the development processes of hosts. These molecules have also shown health-benefiting properties such as antioxidant and anti-inflammatory activities. As such, they are commonly used as pharmaceuticals and nutraceuticals. In plants, flavonoids are assembled through a type III polyketide synthase (PKS) and associated tailoring enzymes such as hydroxylase and glycosyltransferase. Some filamentous fungi also produce flavonoids, such as chlorflavonin from Aspergillus candidus. Chlorflavonin is a novel halogenated flavonoid found in nature reported to possess antifungal and anti-tuberculosis activity. Unlike plants, the biosynthetic process of flavonoids in fungi is not well understood, which has largely hampered our ability to engineer this type of biosynthetic machinery to create new flavonoids. The purpose of this project was to find out whether Cfv1 plays an essential role in chlorflavonin biosynthesis using a gene knockout approach through the Agrobacterium-mediated transformation method. A knockout plasmid, pOKF93, was constructed that harbors a hygromycin resistance gene flanked by the regions upstream and downstream of the cfv1 gene in Aspergillus candidus NRRL 5214. This plasmid was introduced into A. candidus NRRL 5214 through Agrobacterium-mediated transformation. A. candidus NRRL 5214 mutants were selected on Czapek Dox agar plates containing 100 µg/ml hygromycin B and 100 µg/ml cefotaxime. Their products were extracted with methanol and analyzed using High Performance Liquid Chromatography (HPLC). The result showed that knockout of cfv1 has abolished the production of chlorflavonin, indicating that Cfv1 is the dedicated PKS for the assembly of chlorflavonin.
Location
Logan, UT
Start Date
4-11-2023 12:30 PM
End Date
4-11-2023 1:30 PM
Understanding the Role of a Putative Type I Polyketide Synthase in Chlorflavonin Biosynthesis
Logan, UT
Flavonoids are an important class of plant secondary metabolites with a variety of biological functions in the development processes of hosts. These molecules have also shown health-benefiting properties such as antioxidant and anti-inflammatory activities. As such, they are commonly used as pharmaceuticals and nutraceuticals. In plants, flavonoids are assembled through a type III polyketide synthase (PKS) and associated tailoring enzymes such as hydroxylase and glycosyltransferase. Some filamentous fungi also produce flavonoids, such as chlorflavonin from Aspergillus candidus. Chlorflavonin is a novel halogenated flavonoid found in nature reported to possess antifungal and anti-tuberculosis activity. Unlike plants, the biosynthetic process of flavonoids in fungi is not well understood, which has largely hampered our ability to engineer this type of biosynthetic machinery to create new flavonoids. The purpose of this project was to find out whether Cfv1 plays an essential role in chlorflavonin biosynthesis using a gene knockout approach through the Agrobacterium-mediated transformation method. A knockout plasmid, pOKF93, was constructed that harbors a hygromycin resistance gene flanked by the regions upstream and downstream of the cfv1 gene in Aspergillus candidus NRRL 5214. This plasmid was introduced into A. candidus NRRL 5214 through Agrobacterium-mediated transformation. A. candidus NRRL 5214 mutants were selected on Czapek Dox agar plates containing 100 µg/ml hygromycin B and 100 µg/ml cefotaxime. Their products were extracted with methanol and analyzed using High Performance Liquid Chromatography (HPLC). The result showed that knockout of cfv1 has abolished the production of chlorflavonin, indicating that Cfv1 is the dedicated PKS for the assembly of chlorflavonin.