The Co-Optimization of fuels & Engines (Co-Optima) project is a collaboration between nine national laboratories, and industry to develop new fuels and engines, in order to maximize vehicle performance, energy efficiency, and minimize environmental impact. A non-model fungal chassis organism Rhodosporidium toruloides was studied to produce biochemical derived fuels molecules. Rhodosporidium toruloides is oleaginous yeast, capable of accumulating large amounts of lipids. It can utilize both glucose and xylose, which indicates thatit is suitable to produce biofuel molecules from lignocellulosic biomass. Rhodosporidium naturally produces C40 tetraterpenoid carotenoids as red pigment, indicating that the Rhodosporidium native Mevalonate (MVA) pathway could possibly provide important terpene biosynthesis intermediates and precursors, or substrates for producing fuel molecules originating from isoprenoid pathways. Genetic manipulations in Rhodosporidium are also established to do target gene deletion by homologous recombination (HR) and random gene insertion with multiple copy numbers into genomes by non-homologous end-joining (NHEJ). Twenty constructs having genes coding terpene synthase for making C5 terpene alcohols or C10 monoterpene have been transformed to Rhodosporidium, and evaluated for the chemical production by GC-MS. Cineole synthase from Hypoxylon SP. E74060B produced the robust monoterpene compound 1-8-cineole at ~20mg/L level in a dodecane overlay culture. 1-8-Cineole has an observed Research octane number of 99.2 which indicates it is a good high performance biofuel molecule for spark ignition engines.

High performance terpenoid biofuel production by oleaginous yeast Rhodosporidium toruloides

253rd National Meeting (2017)

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