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Tools to make Stachybotrys chartarum genetically amendable: Key to unlocking cryptic biosynthetic gene clusters
Katharina Steinert1, Anna K Atanasoff-Kardjalieff2, Elias Messner2
1Institute of Food Chemistry, University of Münster, Münster, Germany.
Abstract:
The soil and indoor fungus Stachybotrys chartarum can induce respiratory disorders, collectively referred to as stachybotryotoxicosis, owing to its prolific production of diverse bioactive secondary metabolites (SMs) or mycotoxins. Although many of these toxins responsible for the harmful effects on animals and humans have been identified in the genus Stachybotrys, however a number of SMs remain elusive. Through in silico analyses, we have identified 37 polyketide synthase (PKS) genes, highlighting that the chemical profile potential of Stachybotrys is far from being fully explored. Additionally, by leveraging phylogenetic analysis of known SMs produced by non-reducing polyketide synthases (NR-PKS) in other filamentous fungi, we showed that Stachybotrys possesses a rich reservoir of untapped SMs. To unravel natural product biosynthesis in S. chartarum, genetic engineering methods are crucial. For this purpose, we have developed a reliable protocol for the genetic transformation of S. chartarum and applied it to the ScPKS14 biosynthetic gene cluster. This cluster is homologous to the already known Claviceps purpurea CpPKS8 BGC, responsible for the production of ergochromes. While no novel SMs were detected, we successfully applied genetic tools, such as the generation of deletionand overexpression strains of single cluster genes. This toolbox can now be readily employed to unravel not only this particular BGC but also other candidate BGCs present in S. chartarum, making this fungus accessible for genetic engineering.
Insights
Stachybotrys chartarum produces toxins causing respiratory issues. Researchers identified new genes and developed genetic tools to explore its untapped chemical potential for future discoveries.
Area of Science:
- Mycology
- Biochemistry
- Genetics
Background:
- Stachybotrys chartarum is a fungus known for producing mycotoxins that cause respiratory problems (stachybotryotoxicosis).
- While many toxins are identified, a significant number of secondary metabolites (SMs) remain undiscovered.
- In silico analyses suggest a vast, unexplored chemical potential within Stachybotrys, with 37 identified polyketide synthase (PKS) genes.
Purpose of the Study:
- To investigate the unexplored secondary metabolite (SM) biosynthesis in Stachybotrys chartarum.
- To develop and apply genetic engineering tools for unraveling natural product biosynthesis in this fungus.
Main Methods:
- In silico analysis to identify polyketide synthase (PKS) genes.
- Phylogenetic analysis of known SMs produced by non-reducing polyketide synthases (NR-PKS).
- Development of a genetic transformation protocol for S. chartarum and its application to the ScPKS14 gene cluster, including deletion and overexpression studies.
Main Results:
- Identified 37 PKS genes in S. chartarum, indicating significant unexplored biosynthetic potential.
- Established a reliable genetic transformation protocol for S. chartarum.
- Successfully generated deletion and overexpression strains for single genes within the ScPKS14 cluster.
- No novel SMs were detected in this initial study, but the genetic toolbox is now available.
Conclusions:
- Stachybotrys chartarum harbors a rich reservoir of untapped secondary metabolites.
- A functional genetic engineering toolbox has been established for S. chartarum, enabling future research into its biosynthetic gene clusters (BGCs).
- This advancement makes S. chartarum amenable to genetic manipulation for discovering novel compounds.
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