Publications
ThesisJan 2022

Synthesis of fungal secondary metabolites via heterologous expression

Dwars, E
Product Used
Genes
Abstract
In this study, we laid the foundations of a bioinformatic tool to enable for automated gene curation, to facilitate a more correct prediction of fungal gene models. The goal is to employ this tool in the groups of genes (biosynthetic gene clusters, BGCs) coding for enzymes related to the biosynthesis of secondary metabolites (SMs). Correct gene models are crucial for researchers that intend to clone and express BGCs, especially if the pathway of interest is not expressed in a native host under standard laboratory conditions. The gene curation tool can currently split genes to organized blocks representing an exon, and further development will focus on analyses of these blocks to enable automated curation. Additionally, genes from a BGC originating from Aspergillus melleus (AspMel) and genes from a BGC originating from Beauveria bassiana (BeaBas) were expressed in the heterologous host A. oryzae NSAR1. HPLC and LC-MS analysis of the produced and extracted metabolites revealed that the compounds produced by transformants with the AspMel non-reducing polyketide synthase (AME1) alone or with AME1 and tailoring gene enoyl reductase absorb light at 213/214 and 261/260 nm respectively, and compounds of the latter, most likely a previously unidentified compound, have a mass of 168 Da. Additionally, the compounds produced by transformants with AME1 and three tailoring genes (FAD binding protein, Omethyltransferase, and questin oxidase-like) consistently inhibit growth of Escherichia coli and of Bacillus subtilis. The transformants with genes from BeaBas did not show any chemical properties or consistent bioactivity
Product Used
Genes

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