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
| Originalsprog | Engelsk |
|---|---|
| Publikationsdato | 14 okt. 2019 |
| Status | Udgivet - 14 okt. 2019 |
| Begivenhed | SBA2019: Synthetic Biology Australasia Conference 2019 - The Old Customs House, Brisbane, Australien Varighed: 14 okt. 2019 → 16 okt. 2019 https://synbioaustralasia.org/2019-sba-conference/ |
Konference
| Konference | SBA2019 |
|---|---|
| Lokation | The Old Customs House |
| Land/Område | Australien |
| By | Brisbane |
| Periode | 14/10/2019 → 16/10/2019 |
| Internetadresse |