Anaerobic gut fungi are attractive lignocellulose-degrading microbes, yet the enzymatic mechanisms responsible for fungal hydrolysis remain unknown. To discover novel biomass-degrading enzymes and characterize their coordinated expression in fungi, we have implemented methods to sustain an anaerobic fungus in batch culture and analyze its transcriptome via RNAseq under several growth conditions. A new species of gut fungus from the Piromyces genus was isolated from the digestive tract of a horse, and its proliferation was monitored via fermentation gas production. Fungi exhibited high enzymatic reactivity against cellulosic and lignocellulosic substrates (filter paper, reed canary grass), which was repressed in the presence of simple sugars. Through strand-specific RNAseq and use of the TRINITY assembly platform, we were able to assemble novel cellulase genes de novo from >27,000 transcripts without the need for genomic sequence information. We will discuss the coordinated regulation patterns observed for important enzyme families under catabolic regulatory conditions.

Transcriptomic analysis reveals novel lignocellulolytic enzymes from anaerobic gut fungi

245th National Meeting (2013)

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