PT - JOURNAL ARTICLE AU - Junko Yaegashi AU - James Kirby AU - Masakazu Ito AU - Jian Sun AU - Tanmoy Dutta AU - Mona Mirsiaghi AU - Eric R. Sundstrom AU - Alberto Rodriguez AU - Edward Baidoo AU - Deepti Tanjore AU - Todd Pray AU - Kenneth Sale AU - Seema Singh AU - Jay D. Keasling AU - Blake A. Simmons AU - Steven W. Singer AU - Jon K. Magnuson AU - Adam P. Arkin AU - Jeffrey M. Skerker AU - John M. Gladden TI - <em>Rhodosporidium toruloides:</em> A new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts AID - 10.1101/154872 DP - 2017 Jan 01 TA - bioRxiv PG - 154872 4099 - http://biorxiv.org/content/early/2017/06/23/154872.short 4100 - http://biorxiv.org/content/early/2017/06/23/154872.full AB - Background Economical conversion of lignocellulosic biomass into biofuels and bioproducts is central to the establishment of a robust bioeconomy. This requires a conversion host that is able to both efficiently assimilate the major lignocellulose-derived carbon sources and divert their metabolites toward specific bioproducts.Results In this study, the carotenogenic yeast Rhodosporidium toruloides was examined for its ability to convert lignocellulose into two non-native sesquiterpenes with biofuel (bisabolene) and pharmaceutical (amorphadiene) applications. We found that R. toruloides can efficiently convert a mixture of glucose and xylose from hydrolyzed lignocellulose into these bioproducts, and unlike many conventional production hosts, its growth and productivity were enhanced in lignocellulosic hydrolysates relative to purified substrates. This organism was demonstrated to have superior growth in corn stover hydrolysates prepared by two different pretreatment methods, one using a novel biocompatible ionic liquid (IL) choline α-ketoglutarate, which produced 261 mg/L of bisabolene at bench-scale, and the other using an alkaline pretreatment, which produced 680 mg/L of bisabolene in a high gravity fed-batch bioreactor. Interestingly, R. toruloides was also observed to assimilate p-coumaric acid liberated from acylated grass lignin in the IL hydrolysate, a finding we verified with purified substrates. R. toruloides was also able to consume several additional compounds with aromatic motifs similar to lignin monomers, suggesting that this organism may have the metabolic potential to convert depolymerized lignin streams alongside lignocellulosic sugars.Conclusions This study highlights the natural compatibility of R. toruloides with bioprocess conditions relevant to lignocellulosic biorefineries and demonstrates its ability to produce non-native terpenes.ADSamorphadiene synthaseATMTAgrobacterium tumefaciens mediated transformationBISbisabolene synthaseCAPEXcapital expense[Ch][α-Kg]choline α-ketoglutarateCrIcrystallinity indexCSMcomplete supplemental mixtureDIdeionizedGC-MSgas chromatography-mass spectrometryHPLChigh performance liquid chromatographyILionic liquidLBLuria BrothLCAlife cyle analysisODoptical densityOPEXoperating expenseSDsynthetic definedTEAtechnoeconomicXRDX-ray diffractionYNByeast nitrogen baseYPDyeast peptone dextrose