Heterologous expression of the fungal polyketide bikaverin and effects of PPTase co-expression

Publikation: Konferencebidrag uden forlag/tidsskriftPosterForskning

Resumé

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
OriginalsprogEngelsk
Publikationsdato14 okt. 2019
StatusUdgivet - 14 okt. 2019
BegivenhedSBA2019: Synthetic Biology Australasia Conference 2019 - The Old Customs House, Brisbane, Australien
Varighed: 14 okt. 201916 okt. 2019
https://synbioaustralasia.org/2019-sba-conference/

Konference

KonferenceSBA2019
LokationThe Old Customs House
LandAustralien
ByBrisbane
Periode14/10/201916/10/2019
Internetadresse

Fingerprint

polyketides
biochemical pathways
Saccharomyces cerevisiae
chemical analysis
plasmids
pigments
extracts
enzymes
Fusarium verticillioides
tandem mass spectrometry

Citer dette

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title = "Heterologous expression of the fungal polyketide bikaverin and effects of PPTase co-expression",
abstract = "The biosynthetic pathway of the Fusarium verticillioides polyketide bikaverin (PKS16) was reconstructedutilizing 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 ofthe biosynthetic pathway was validated and the red pigment of bikaverin was produced. Whileheterologous 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 polyketidesynthase (PKS) from an inactive apo-conformation to the active holo-conformation. This study elucidatesthe effect of seven different PPTases on the overall yield of bikaverin and determines the most efficient tobe FvPPT, a PPTase also originating from Fusarium verticillioides, yielding 58 mg/L bikaverin. This couldindicate an evolutionary advantage when pairing PPTases and pathways sharing the same origin, inheterologous expression systems",
author = "Pedersen, {Tobias Bruun} and Gardiner, {Donald Max} and S{\o}rensen, {Jens Laurids}",
year = "2019",
month = "10",
day = "14",
language = "English",
note = "SBA2019 : Synthetic Biology Australasia Conference 2019, SBA2019 ; Conference date: 14-10-2019 Through 16-10-2019",
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Heterologous expression of the fungal polyketide bikaverin and effects of PPTase co-expression. / Pedersen, Tobias Bruun; Gardiner, Donald Max; Sørensen, Jens Laurids.

2019. Poster præsenteret på SBA2019, Brisbane, Australien.

Publikation: Konferencebidrag uden forlag/tidsskriftPosterForskning

TY - CONF

T1 - Heterologous expression of the fungal polyketide bikaverin and effects of PPTase co-expression

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AU - Gardiner, Donald Max

AU - Sørensen, Jens Laurids

PY - 2019/10/14

Y1 - 2019/10/14

N2 - The biosynthetic pathway of the Fusarium verticillioides polyketide bikaverin (PKS16) was reconstructedutilizing 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 ofthe biosynthetic pathway was validated and the red pigment of bikaverin was produced. Whileheterologous 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 polyketidesynthase (PKS) from an inactive apo-conformation to the active holo-conformation. This study elucidatesthe effect of seven different PPTases on the overall yield of bikaverin and determines the most efficient tobe FvPPT, a PPTase also originating from Fusarium verticillioides, yielding 58 mg/L bikaverin. This couldindicate an evolutionary advantage when pairing PPTases and pathways sharing the same origin, inheterologous expression systems

AB - The biosynthetic pathway of the Fusarium verticillioides polyketide bikaverin (PKS16) was reconstructedutilizing 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 ofthe biosynthetic pathway was validated and the red pigment of bikaverin was produced. Whileheterologous 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 polyketidesynthase (PKS) from an inactive apo-conformation to the active holo-conformation. This study elucidatesthe effect of seven different PPTases on the overall yield of bikaverin and determines the most efficient tobe FvPPT, a PPTase also originating from Fusarium verticillioides, yielding 58 mg/L bikaverin. This couldindicate an evolutionary advantage when pairing PPTases and pathways sharing the same origin, inheterologous expression systems

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