Development of Genetic Dereplication Strains in <i>Aspergillus nidulans</i> Results in the Discovery of Aspercryptin

Chiang, Yi‐Ming; Ahuja, Manmeet; Oakley, C. Elizabeth; Entwistle, Ruth; Asokan, Anabanadam; Zutz, Christoph; Wang, Clay C. C.; Oakley, Berl R.

doi:10.1002/anie.201507097

Cited by 157 publications

(141 citation statements)

References 31 publications

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“…We have previously developed methods for deleting entire A. nidulans SM clusters and we have used these methods to engineer a strain (LO8030) in which the SM clusters responsible for the biosynthesis of the following major SMs are deleted: sterigmatocystin, the emericellamides, asperfuranone, monodictyphenone, terrequinone, F9775A and B, asperthecin and both portions of the split SM cluster that produces austinol and dehydroaustinol (Chiang et al, 2016). These deletions greatly reduce the SM background allowing SMs from other clusters to be detected more easily.…”

Section: Deletion Of An8694 In a Genetic Dereplication Strain Resultsmentioning

confidence: 99%

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Oakley

Ahuja

Sun

et al. 2016

Molecular Microbiology

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Summary Fungal secondary metabolites (SMs) are extremely important in medicine and agriculture, but regulation of their biosynthesis is incompletely understood. We have developed a genetic screen in Aspergillus nidulans for negative regulators of fungal SM gene clusters and we have used this screen to isolate mutations that upregulate transcription of the non-ribosomal peptide synthetase gene required for nidulanin A biosynthesis. Several of these mutations are allelic and we have identified the mutant gene by genome sequencing. The gene, which we designate mcrA, is conserved but uncharacterized, and it encodes a putative transcription factor. Metabolite profiles of mcrA deletant, mcrA overexpressing, and parental strains reveal that mcrA regulates at least ten SM gene clusters. Deletion of mcrA stimulates SM production even in strains carrying a deletion of the SM regulator laeA, and deletion of mcrA homologs in Aspergillus terreus and Penicillum canescens alters the secondary metabolite profile of these organisms. Deleting mcrA in a genetic dereplication strain has allowed us to discover two novel compounds as well as an antibiotic not known to be produced by A. nidulans. Deletion of mcrA upregulates transcription of hundreds of genes including many that are involved in secondary metabolism, while downregulating a smaller number of genes.

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Section: Deletion Of An8694 In a Genetic Dereplication Strain Resultsmentioning

confidence: 99%

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Oakley

Ahuja

Sun

et al. 2016

Molecular Microbiology

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“…nidulans LO7890 (a gift from Prof Berl Oakley, University of Kansas) was used as the heterologous host for expression of elcA. A. nidulans LO7890 is similar to LO8030 as described by Chiang et al (Chiang et al, 2016), which has eight secondary metabolite biosynthetic gene clusters deleted, except that LO7890 still contains the asperthecin gene cluster (only seven gene clusters are deleted). elcA together with the A. nidulans alcA promoter was cloned into pYFAC, a plasmid containing the A. nidulans plasmid replicator AMA1 (Aleksenko and Clutterbuck, 1996) and yeast CEN/ARS, by yeast transformationmediated homologous recombination.…”

Section: Heterologous Expression Of Elca In Aspergillus Nidulansmentioning

confidence: 99%

Functional genomics‐guided discovery of a light‐activated phytotoxin in the wheat pathogen Parastagonospora nodorum via pathway activation

Chooi

Zhang

et al. 2017

Environmental Microbiology

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Parastagonospora nodorum is an important pathogen of wheat. The contribution of secondary metabolites to this pathosystem is poorly understood. A biosynthetic gene cluster (SNOG_08608-08616) has been shown to be upregulated during the late stage of P. nodorum wheat leaf infection. The gene cluster shares several homologues with the Cercospora nicotianae CTB gene cluster encoding the biosynthesis of cercosporin. Activation of the gene cluster by overexpression (OE) of the transcription factor gene (SNOG_08609) in P. nodorum resulted in the production of elsinochrome C, a perelyenequinone phytotoxin structurally similar to cercosporin. Heterologous expression of the polyketide synthase gene elcA from the gene cluster in Aspergillus nidulans resulted in the production of the polyketide precursor nortoralactone common to the cercosporin pathway. Elsinochrome C could be detected on wheat leaves infected with P. nodorum, but not in the elcA disruption mutant. The compound was shown to exhibit necrotic activity on wheat leaves in a light-dependent manner. Wheat seedling infection assays showed that ΔelcA exhibited reduced virulence compared with wild type, while infection by an OE strain overproducing elsinochrome C resulted in larger lesions on leaves. These data provided evidence that elsinochrome C contributes to the virulence of P. nodorum against wheat.

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“…However, the enzyme responsible for the aldol reaction to construct the unique tricyclo[6.2.2.0]dodecane scaffold in 1 and the related betaenone A ( 5 ) from P. betae remained enigmatic. Herein, to further shed light into the biosynthesis of 1 , we employed a similar heterologous pathway reconstruction approach in A. nidulans LO7890 by using a hybrid yeast‐fungal artificial chromosome (pYFAC) expression system . Co‐expression of the polyketide synthase gene sthA and the partnering trans ‐ER gene sthE in A. nidulans led to the production of dehydroprobetaenone I 2 (Figure ), which corresponds to co‐expression of bet1 and bet3 (homologous to sthA and E , respectively) in A. oryzae .…”

Section: Figurementioning

confidence: 99%

Biosynthesis of a Tricyclo[6.2.2.0^2,7]dodecane System by a Berberine Bridge Enzyme‐Like Aldolase

Wei

et al. 2019

Chemistry A European J

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The aldol reaction is one of the most fundamental stereocontrolled carbon–carbon bond‐forming reactions and is mainly catalyzed by aldolases in nature. Despite the fact that the aldol reaction has been widely proposed to be involved in fungal secondary metabolite biosynthesis, a dedicated aldolase that catalyzes stereoselective aldol reactions has only rarely been reported in fungi. Herein, we activated a cryptic polyketide biosynthetic gene cluster that was upregulated in the fungal wheat pathogen Parastagonospora nodorum during plant infection; this resulted in the production of the phytotoxic stemphyloxin II (1). Through heterologous reconstruction of the biosynthetic pathway and in vitro assay by using cell‐free lysate from Aspergillus nidulans, we demonstrated that a berberine bridge enzyme (BBE)‐like protein SthB catalyzes an intramolecular aldol reaction to establish the bridged tricyclo[6.2.2.02,7]dodecane skeleton in the post‐assembly tailoring step. The characterization of SthB as an aldolase enriches the catalytic toolbox of classic reactions and the functional diversities of the BBE superfamily of enzymes.

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Development of Genetic Dereplication Strains in Aspergillus nidulans Results in the Discovery of Aspercryptin

Cited by 157 publications

References 31 publications

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Functional genomics‐guided discovery of a light‐activated phytotoxin in the wheat pathogen Parastagonospora nodorum via pathway activation

Biosynthesis of a Tricyclo[6.2.2.0^2,7]dodecane System by a Berberine Bridge Enzyme‐Like Aldolase

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Development of Genetic Dereplication Strains in Aspergillus nidulans Results in the Discovery of Aspercryptin

Cited by 157 publications

References 31 publications

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Discovery of McrA, a master regulator of Aspergillus secondary metabolism

Functional genomics‐guided discovery of a light‐activated phytotoxin in the wheat pathogen Parastagonospora nodorum via pathway activation

Biosynthesis of a Tricyclo[6.2.2.02,7]dodecane System by a Berberine Bridge Enzyme‐Like Aldolase

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Biosynthesis of a Tricyclo[6.2.2.0^2,7]dodecane System by a Berberine Bridge Enzyme‐Like Aldolase