Abstract
The biosynthetic pathway of the Fusarium verticillioides polyketide bikaverin (PKS16) was reconstructed
utilizing a plasmid-based system and heterologously expressed in Saccharomyces cerevisiae BY4743.
Through NGS-sequencing and subsequent HPLC MS-MS analysis of chemical extracts, the reconstruction of
the biosynthetic pathway was validated and the red pigment of bikaverin was produced. While
heterologous expressing fungal polyketides in S. cerevisiae the co-expression of a helper enzyme, a 4-
phosphopantetheinyltransferase (PPTase), is necessary for transforming the ACP-domain of the polyketide
synthase (PKS) from an inactive apo-conformation to the active holo-conformation. This study elucidates
the effect of seven different PPTases on the overall yield of bikaverin and determines the most efficient to
be FvPPT, a PPTase also originating from Fusarium verticillioides, yielding 58 mg/L bikaverin. This could
indicate an evolutionary advantage when pairing PPTases and pathways sharing the same origin, in
heterologous expression systems
utilizing a plasmid-based system and heterologously expressed in Saccharomyces cerevisiae BY4743.
Through NGS-sequencing and subsequent HPLC MS-MS analysis of chemical extracts, the reconstruction of
the biosynthetic pathway was validated and the red pigment of bikaverin was produced. While
heterologous expressing fungal polyketides in S. cerevisiae the co-expression of a helper enzyme, a 4-
phosphopantetheinyltransferase (PPTase), is necessary for transforming the ACP-domain of the polyketide
synthase (PKS) from an inactive apo-conformation to the active holo-conformation. This study elucidates
the effect of seven different PPTases on the overall yield of bikaverin and determines the most efficient to
be FvPPT, a PPTase also originating from Fusarium verticillioides, yielding 58 mg/L bikaverin. This could
indicate an evolutionary advantage when pairing PPTases and pathways sharing the same origin, in
heterologous expression systems
| Original language | English |
|---|---|
| Publication date | 14 Oct 2019 |
| Publication status | Published - 14 Oct 2019 |
| Event | SBA2019: Synthetic Biology Australasia Conference 2019 - The Old Customs House, Brisbane, Australia Duration: 14 Oct 2019 → 16 Oct 2019 https://synbioaustralasia.org/2019-sba-conference/ |
Conference
| Conference | SBA2019 |
|---|---|
| Location | The Old Customs House |
| Country/Territory | Australia |
| City | Brisbane |
| Period | 14/10/2019 → 16/10/2019 |
| Internet address |