Membrane transporters are critical gatekeepers of the cell that control the uptake of needed substrates and the secretion of valuable fuel products. Industrial workhorses, however, frequently lack the transporters to effectively use substrates from renewable plant biomass, and secrete toxic fuel products from the intracellular environment. For instance, Saccharomyces is unable to natively use the pentose sugars from crude biomass in engineered fuel and energy producing pathways due, in part, to a lack of uptake mechanism. In contrast, anaerobic fungi from the guts of large herbivores are able to thrive on an array of sugars resulting from lignocellulose hydrolysis. Here, we present a systems-level strategy integrating transcriptomics and phenotype to mine this diverse phylum for novel sugar transporters for improved bioprocessing. Three new gut fungal species, representing three distinct genera, were examined with RNAseq to generate de novo transcriptomes containing hundreds of putative sugar transporters. These transporters were classified on the basis of homology into families as described in the Transporter Classification Database (TCDB) and assigned a putative substrate. Substrate predictions were subsequently validated by comparing phenotype against metabolic bottlenecks and transporter candidates to reveal unique transporters putatively responsible for sugar uptake. Transcriptomic sequencing also revealed a number of multidrug transporter candidates which may further increase productivity by increasing fuel tolerance via product secretion. In this talk, I expand on this workflow for general transcriptome mining and discuss some of the more promising candidates.

Novel gut fungal transporters for improved fuel and energy production

251st National Meeting (2016)

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