Lokhorst, Iris, I (2022) Novel Biocatalysts and Secondary Metabolites from Fungi. Master's Internship Report, Pharmacy.
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Abstract
This study aims to explore uncharacterized biosynthetic enzymes from the understudied group of lichen-forming fungi, via expression in a heterologous filamentous host (Aspergillus oryzae NSAR1). Using this approach, we successfully cloned seven genes from five organisms. Two enzymes, both putative dimethylallyltryptophan synthases (DMATS) (from Ramalina intermedia and Acarospora strigata) showed activity in in vivo experiments. LC-MS results indicate that they can attach a dimethylallyl group not to tryptophan, but to at least two other substrates: tyrosine and N-acetyl-tyrosine. The overexpression of another gene from R. intermedia resulted in altered morphology but appears to have no effect on the metabolome. Furthermore, the groundwork for a series of co-cultivation experiments is laid with the aim to activate and investigate otherwise silent metabolite pathways in fungi. Several visually interesting fungi-fungi matches have been set up, and preliminary LC-MS analysis showed a handful of new and unidentified metabolites. Finally, we investigate the metabolic potential of a wild isolate via anti-microbial assays, which showed anti-Gram+ activity. These experiments merely scratch the surface of the plethora of unannotated fungal biosynthetic enzymes. Further analysis will be required to elucidate the structure of the newly discovered small molecules and to identify the genes that are activated upon fungi-fungi contact.
| Item Type: | Thesis (Master's Internship Report) |
|---|---|
| Supervisor name: | Haslinger, K. and Iacovelli, R. |
| Degree programme: | Pharmacy |
| Thesis type: | Master's Internship Report |
| Language: | English |
| Date Deposited: | 31 Mar 2023 11:54 |
| Last Modified: | 31 Mar 2023 11:54 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/29042 |
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