Identification of the first steps in phenalenone pigment biosynthesis in Fusarium solani

Publikation: Bidrag til bog/antologi/rapport/konference proceedingKonferenceabstrakt i proceedingForskning

Resumé

Most Fusarium species produce bikaverin and aurofusarin for mycelium pigmentation and fusarubins for perithecial pigmentation [1]. However, Fusarium solani produces fusarubins during mycelial growth and another unknown pigment during sexual reproduction. This unknown pigment is predicted to be biosynthesized by a non-reducing polyketide synthase (PKS35 = PKSN [2]). PKS35 encodes a 2106 aa protein and share 52% identity to PhnA from P. herquei, which catalyzes the first step of the herqueinone biosynthetic pathway [3]. Genetic analyses of the PKS35 gene cluster suggests that it contains eleven genes, of which four are present in the herqueinone cluster. Herqueinone contains tricyclic phenalenone core ring structure cyclized by a FAD-dependent monooxygenase. To unravel the biosynthetic products in the PKS35 pathway the intron stripped PKS was cloned and put under control of a galactose inducible promoter in a 2µ vector, which was transformed into a Saccharomyces cerevisiae strain co-expressing a Sfp-Type 4′-Phosphopantetheinyl Transferase (Ppt1). The transformed yeast strain was cultivated under induced conditions in liquid cultures for five days and before production of secondary metabolites
was analyzed by high-resolution mass spectrometry (HRMS). The results showed that the yeast strain was able to produce prephenalenone, which is also the first step of the herqueinone pathway. We also detected dehydroxyprephenalenone, which is formed through spontaneous dehydration [3]. Through heterologous expression of the other genes in the cluster, we will follow the biosynthetic route and ultimately unravel the entire pathway of the perithecial pigment of F. solani.
[1] Studt L, Wiemann P, Kleigrewe K et al (2012): Appl Environ Microbiol, 78: 4468-4480.
[2] Graziani S, Vasnier C, Daboussi MJ (2004): Appl Environ Microbiol, 70: 2984-2988.
[3] Gao SS, Duan A, Xu W et al (2016): J Am Chem Soc, 138: 4249-4259.
OriginalsprogEngelsk
TitelEFS14 - EUROPEAN FUSARIUM SEMINAR : Poster presentations
Publikationsdato2018
Sider100
ArtikelnummerPP-78
StatusUdgivet - 2018
BegivenhedEuropean Fusarium Seminar 14 - Tulln, Østrig
Varighed: 8 apr. 201811 apr. 2018

Konference

KonferenceEuropean Fusarium Seminar 14
LokationTulln
LandØstrig
Periode08/04/201811/04/2018

Fingerprint

Fusarium solani
pigments
biosynthesis
multigene family
pigmentation
yeasts
polyketide synthases
sexual reproduction
transferases
galactose
Fusarium
mycelium
biochemical pathways
introns
Saccharomyces cerevisiae
promoter regions
mass spectrometry
liquids
genes
proteins

Citer dette

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title = "Identification of the first steps in phenalenone pigment biosynthesis in Fusarium solani",
abstract = "Most Fusarium species produce bikaverin and aurofusarin for mycelium pigmentation and fusarubins for perithecial pigmentation [1]. However, Fusarium solani produces fusarubins during mycelial growth and another unknown pigment during sexual reproduction. This unknown pigment is predicted to be biosynthesized by a non-reducing polyketide synthase (PKS35 = PKSN [2]). PKS35 encodes a 2106 aa protein and share 52{\%} identity to PhnA from P. herquei, which catalyzes the first step of the herqueinone biosynthetic pathway [3]. Genetic analyses of the PKS35 gene cluster suggests that it contains eleven genes, of which four are present in the herqueinone cluster. Herqueinone contains tricyclic phenalenone core ring structure cyclized by a FAD-dependent monooxygenase. To unravel the biosynthetic products in the PKS35 pathway the intron stripped PKS was cloned and put under control of a galactose inducible promoter in a 2µ vector, which was transformed into a Saccharomyces cerevisiae strain co-expressing a Sfp-Type 4′-Phosphopantetheinyl Transferase (Ppt1). The transformed yeast strain was cultivated under induced conditions in liquid cultures for five days and before production of secondary metaboliteswas analyzed by high-resolution mass spectrometry (HRMS). The results showed that the yeast strain was able to produce prephenalenone, which is also the first step of the herqueinone pathway. We also detected dehydroxyprephenalenone, which is formed through spontaneous dehydration [3]. Through heterologous expression of the other genes in the cluster, we will follow the biosynthetic route and ultimately unravel the entire pathway of the perithecial pigment of F. solani.[1] Studt L, Wiemann P, Kleigrewe K et al (2012): Appl Environ Microbiol, 78: 4468-4480.[2] Graziani S, Vasnier C, Daboussi MJ (2004): Appl Environ Microbiol, 70: 2984-2988.[3] Gao SS, Duan A, Xu W et al (2016): J Am Chem Soc, 138: 4249-4259.",
author = "Nielsen, {Mikkel Rank} and Pedersen, {Tobias Bruun} and Rasmus Wollenberg and S{\o}ndergaard, {Teis Esben} and S{\o}rensen, {Jens Laurids}",
year = "2018",
language = "English",
pages = "100",
booktitle = "EFS14 - EUROPEAN FUSARIUM SEMINAR",

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Nielsen, MR, Pedersen, TB, Wollenberg, R, Søndergaard, TE & Sørensen, JL 2018, Identification of the first steps in phenalenone pigment biosynthesis in Fusarium solani. i EFS14 - EUROPEAN FUSARIUM SEMINAR: Poster presentations., PP-78, s. 100, European Fusarium Seminar 14, Østrig, 08/04/2018.

Identification of the first steps in phenalenone pigment biosynthesis in Fusarium solani. / Nielsen, Mikkel Rank; Pedersen, Tobias Bruun; Wollenberg, Rasmus; Søndergaard, Teis Esben; Sørensen, Jens Laurids.

EFS14 - EUROPEAN FUSARIUM SEMINAR: Poster presentations. 2018. s. 100 PP-78.

Publikation: Bidrag til bog/antologi/rapport/konference proceedingKonferenceabstrakt i proceedingForskning

TY - ABST

T1 - Identification of the first steps in phenalenone pigment biosynthesis in Fusarium solani

AU - Nielsen, Mikkel Rank

AU - Pedersen, Tobias Bruun

AU - Wollenberg, Rasmus

AU - Søndergaard, Teis Esben

AU - Sørensen, Jens Laurids

PY - 2018

Y1 - 2018

N2 - Most Fusarium species produce bikaverin and aurofusarin for mycelium pigmentation and fusarubins for perithecial pigmentation [1]. However, Fusarium solani produces fusarubins during mycelial growth and another unknown pigment during sexual reproduction. This unknown pigment is predicted to be biosynthesized by a non-reducing polyketide synthase (PKS35 = PKSN [2]). PKS35 encodes a 2106 aa protein and share 52% identity to PhnA from P. herquei, which catalyzes the first step of the herqueinone biosynthetic pathway [3]. Genetic analyses of the PKS35 gene cluster suggests that it contains eleven genes, of which four are present in the herqueinone cluster. Herqueinone contains tricyclic phenalenone core ring structure cyclized by a FAD-dependent monooxygenase. To unravel the biosynthetic products in the PKS35 pathway the intron stripped PKS was cloned and put under control of a galactose inducible promoter in a 2µ vector, which was transformed into a Saccharomyces cerevisiae strain co-expressing a Sfp-Type 4′-Phosphopantetheinyl Transferase (Ppt1). The transformed yeast strain was cultivated under induced conditions in liquid cultures for five days and before production of secondary metaboliteswas analyzed by high-resolution mass spectrometry (HRMS). The results showed that the yeast strain was able to produce prephenalenone, which is also the first step of the herqueinone pathway. We also detected dehydroxyprephenalenone, which is formed through spontaneous dehydration [3]. Through heterologous expression of the other genes in the cluster, we will follow the biosynthetic route and ultimately unravel the entire pathway of the perithecial pigment of F. solani.[1] Studt L, Wiemann P, Kleigrewe K et al (2012): Appl Environ Microbiol, 78: 4468-4480.[2] Graziani S, Vasnier C, Daboussi MJ (2004): Appl Environ Microbiol, 70: 2984-2988.[3] Gao SS, Duan A, Xu W et al (2016): J Am Chem Soc, 138: 4249-4259.

AB - Most Fusarium species produce bikaverin and aurofusarin for mycelium pigmentation and fusarubins for perithecial pigmentation [1]. However, Fusarium solani produces fusarubins during mycelial growth and another unknown pigment during sexual reproduction. This unknown pigment is predicted to be biosynthesized by a non-reducing polyketide synthase (PKS35 = PKSN [2]). PKS35 encodes a 2106 aa protein and share 52% identity to PhnA from P. herquei, which catalyzes the first step of the herqueinone biosynthetic pathway [3]. Genetic analyses of the PKS35 gene cluster suggests that it contains eleven genes, of which four are present in the herqueinone cluster. Herqueinone contains tricyclic phenalenone core ring structure cyclized by a FAD-dependent monooxygenase. To unravel the biosynthetic products in the PKS35 pathway the intron stripped PKS was cloned and put under control of a galactose inducible promoter in a 2µ vector, which was transformed into a Saccharomyces cerevisiae strain co-expressing a Sfp-Type 4′-Phosphopantetheinyl Transferase (Ppt1). The transformed yeast strain was cultivated under induced conditions in liquid cultures for five days and before production of secondary metaboliteswas analyzed by high-resolution mass spectrometry (HRMS). The results showed that the yeast strain was able to produce prephenalenone, which is also the first step of the herqueinone pathway. We also detected dehydroxyprephenalenone, which is formed through spontaneous dehydration [3]. Through heterologous expression of the other genes in the cluster, we will follow the biosynthetic route and ultimately unravel the entire pathway of the perithecial pigment of F. solani.[1] Studt L, Wiemann P, Kleigrewe K et al (2012): Appl Environ Microbiol, 78: 4468-4480.[2] Graziani S, Vasnier C, Daboussi MJ (2004): Appl Environ Microbiol, 70: 2984-2988.[3] Gao SS, Duan A, Xu W et al (2016): J Am Chem Soc, 138: 4249-4259.

UR - https://www.efs14.at/pub/EFS14_Abstractbook_V1.7.pdf

M3 - Conference abstract in proceeding

SP - 100

BT - EFS14 - EUROPEAN FUSARIUM SEMINAR

ER -