Early-diverging anaerobic fungi resident in the digestive tracts of large herbivores are attractive platforms for biotechnology due to their robust ability to degrade untreated lignocellulosic feedstocks. Anaerobic fungi secrete the largest known fungal array of biomass-degrading enzymes, which allow for robust degradation of diverse agricultural, food, and forestry wastes independent of substrate composition. However, there are no tools available to permanently manipulate these fungi and exploit their abilities for enzyme production or consolidated bioprocessing. Here, I present our ongoing efforts to engineer anaerobic fungi. Anaerobic fungi are naturally competent for portions of their life cycle, which can be exploited to introduce heterologous DNA. Using a characterized enolase promoter, we demonstrate heterologous expression of a selection marker enabling library-based screening approaches to construct the first stable anaerobic fungal plasmid. We also establish that linear DNA can be recombined into gut fungal genomes to propagate stable gene modifications. Anaerobic fungi also display epigenetic regulation that can be controlled to increase biomass-degrading enzyme secretion. This growing toolbox of genetic and epigenetic strategies can be readily generalized to other non-model systems, and enables metabolic engineering opportunities in anaerobic fungi for the first time.<br/>

Tool development to manipulate early-branching anaerobic fungi as a platform for biotechnology

2019 Spring National Meeting

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