Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds

ABSTRACT Advances in genome sequencing have revitalized natural product discovery efforts, revealing the untapped biosynthetic potential of fungi. While the volume of genomic data continues to expand, discovery efforts are slowed due to the time-consuming nature of experiments required to characteri...

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Autores principales:	Lindsay K. Caesar, Matthew T. Robey, Michael Swyers, Md N. Islam, Rosa Ye, Purav P. Vagadia, Gary E. Schiltz, Paul M. Thomas, Chengcang C. Wu, Neil L. Kelleher, Nancy P. Keller, Jin Woo Bok
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	IDO heterologous expression natural products Aspergillus terreus Aspergillus nidulans genome mining Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0c3cdc60038744a98df4944edd8388aa
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spelling	oai:doaj.org-article:0c3cdc60038744a98df4944edd8388aa2021-11-15T15:56:44ZHeterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds10.1128/mBio.01691-202150-7511https://doaj.org/article/0c3cdc60038744a98df4944edd8388aa2020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01691-20https://doaj.org/toc/2150-7511ABSTRACT Advances in genome sequencing have revitalized natural product discovery efforts, revealing the untapped biosynthetic potential of fungi. While the volume of genomic data continues to expand, discovery efforts are slowed due to the time-consuming nature of experiments required to characterize new molecules. To direct efforts toward uncharacterized biosynthetic gene clusters most likely to encode novel chemical scaffolds, we took advantage of comparative metabolomics and heterologous gene expression using fungal artificial chromosomes (FACs). By linking mass spectral profiles with structural clues provided by FAC-encoded gene clusters, we targeted a compound originating from an unusual gene cluster containing an indoleamine 2,3-dioxygenase (IDO). With this approach, we isolate and characterize R and S forms of the new molecule terreazepine, which contains a novel chemical scaffold resulting from cyclization of the IDO-supplied kynurenine. The discovery of terreazepine illustrates that FAC-based approaches targeting unusual biosynthetic machinery provide a promising avenue forward for targeted discovery of novel scaffolds and their biosynthetic enzymes, and it also represents another example of a biosynthetic gene cluster “repurposing” a primary metabolic enzyme to diversify its secondary metabolite arsenal. IMPORTANCE Here, we provide evidence that Aspergillus terreus encodes a biosynthetic gene cluster containing a repurposed indoleamine 2,3-dioxygenase (IDO) dedicated to secondary metabolite synthesis. The discovery of this neofunctionalized IDO not only enabled discovery of a new compound with an unusual chemical scaffold but also provided insight into the numerous strategies fungi employ for diversifying and protecting themselves against secondary metabolites. The observations in this study set the stage for further in-depth studies into the function of duplicated IDOs present in fungal biosynthetic gene clusters and presents a strategy for accessing the biosynthetic potential of gene clusters containing duplicated primary metabolic genes.Lindsay K. CaesarMatthew T. RobeyMichael SwyersMd N. IslamRosa YePurav P. VagadiaGary E. SchiltzPaul M. ThomasChengcang C. WuNeil L. KelleherNancy P. KellerJin Woo BokAmerican Society for MicrobiologyarticleIDOheterologous expressionnatural productsAspergillus terreusAspergillus nidulansgenome miningMicrobiologyQR1-502ENmBio, Vol 11, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	IDO heterologous expression natural products Aspergillus terreus Aspergillus nidulans genome mining Microbiology QR1-502
spellingShingle	IDO heterologous expression natural products Aspergillus terreus Aspergillus nidulans genome mining Microbiology QR1-502 Lindsay K. Caesar Matthew T. Robey Michael Swyers Md N. Islam Rosa Ye Purav P. Vagadia Gary E. Schiltz Paul M. Thomas Chengcang C. Wu Neil L. Kelleher Nancy P. Keller Jin Woo Bok Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
description	ABSTRACT Advances in genome sequencing have revitalized natural product discovery efforts, revealing the untapped biosynthetic potential of fungi. While the volume of genomic data continues to expand, discovery efforts are slowed due to the time-consuming nature of experiments required to characterize new molecules. To direct efforts toward uncharacterized biosynthetic gene clusters most likely to encode novel chemical scaffolds, we took advantage of comparative metabolomics and heterologous gene expression using fungal artificial chromosomes (FACs). By linking mass spectral profiles with structural clues provided by FAC-encoded gene clusters, we targeted a compound originating from an unusual gene cluster containing an indoleamine 2,3-dioxygenase (IDO). With this approach, we isolate and characterize R and S forms of the new molecule terreazepine, which contains a novel chemical scaffold resulting from cyclization of the IDO-supplied kynurenine. The discovery of terreazepine illustrates that FAC-based approaches targeting unusual biosynthetic machinery provide a promising avenue forward for targeted discovery of novel scaffolds and their biosynthetic enzymes, and it also represents another example of a biosynthetic gene cluster “repurposing” a primary metabolic enzyme to diversify its secondary metabolite arsenal. IMPORTANCE Here, we provide evidence that Aspergillus terreus encodes a biosynthetic gene cluster containing a repurposed indoleamine 2,3-dioxygenase (IDO) dedicated to secondary metabolite synthesis. The discovery of this neofunctionalized IDO not only enabled discovery of a new compound with an unusual chemical scaffold but also provided insight into the numerous strategies fungi employ for diversifying and protecting themselves against secondary metabolites. The observations in this study set the stage for further in-depth studies into the function of duplicated IDOs present in fungal biosynthetic gene clusters and presents a strategy for accessing the biosynthetic potential of gene clusters containing duplicated primary metabolic genes.
format	article
author	Lindsay K. Caesar Matthew T. Robey Michael Swyers Md N. Islam Rosa Ye Purav P. Vagadia Gary E. Schiltz Paul M. Thomas Chengcang C. Wu Neil L. Kelleher Nancy P. Keller Jin Woo Bok
author_facet	Lindsay K. Caesar Matthew T. Robey Michael Swyers Md N. Islam Rosa Ye Purav P. Vagadia Gary E. Schiltz Paul M. Thomas Chengcang C. Wu Neil L. Kelleher Nancy P. Keller Jin Woo Bok
author_sort	Lindsay K. Caesar
title	Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
title_short	Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
title_full	Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
title_fullStr	Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
title_full_unstemmed	Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds
title_sort	heterologous expression of the unusual terreazepine biosynthetic gene cluster reveals a promising approach for identifying new chemical scaffolds
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/0c3cdc60038744a98df4944edd8388aa
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Heterologous Expression of the Unusual Terreazepine Biosynthetic Gene Cluster Reveals a Promising Approach for Identifying New Chemical Scaffolds

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