Antibiotic resistance has arisen in a significant number of human pathogens, antibiotic discovery and development is, however, currently not keeping pace. This has lead to the reinvestigation of some naturally produced antibiotic compounds which act in a manner that avoids common resistance mechanisms. Pleuromutilin is one such compound with activity against bacteria such as Methicillin Resistant Staphylococcus aureus (MRSA). Pleuromutilin is generally synthesised at a low titre by its native producer Clitopilus passeckerianus and as such research into its biosynthesis may enable yield increases. This project has taken a multifaceted approach to manipulate the Pleuromutilin biosynthetic gene cluster in a variety of fungal organisms. Within C.passeckerianus gene silencing is being used to interfere with specific stages of biosynthesis, silenced transformed lines have been analysed both chemically, to determine the impact of silencing on the metabolome of the organism, and via QRT-PCR, to determine the impact of silencing on the transcription of the gene cluster. Heterologous expression of the entire gene cluster is being evaluated and analysed in the model basidiomycete Coprinopsis cinerea. Engineered expression of the biosynthetic genes is being performed in the industrially relevant ascomycete Aspergillus oryzae to build the pathway in a stepwise manner. Progress in each of these areas will be presented.
Full conference title:
27th Fungal Genetics Conference
- FGC 27th (2013)