Cutaneotrichosporon oleaginosus, previously known as Cryptococcus curvatus, is a non-model oleaginous yeast known for its ability to metabolize many alternative substrates, including xylose, and toxic lignocellulosic hydrolysate inhibitors such as 5-hydroxymethylfurfural and furfural. We discovered C. oleaginosus also tolerates and metabolizes lignin-derived aromatics, highlighting this organisms’ potential to utilize all components of lignocellulosic biomass. C. oleaginosus is able to fully metabolize phenol, 4-hydroxybenzoic acid (pHBA), and resorcinol as sole carbon sources, as well as co-utilization with glucose and xylose. When buffered, cells also metabolize p-coumarate robustly and to a lesser degree ferulate and syringate as sole carbon sources. In a simple batch flask culture, C. oleaginosus is able to accumulate nearly 50% of its biomass as lipids while utilizing pHBA, p-coumarate, and resorcinol. Optimizing fed-batch feeding strategies increased lipid accumulation to over 69%, the second highest value in literature for this organism to date. Transcriptomic analyses were used to elucidate aromatic metabolic pathways in C. oleaginosus and improve the existing genome annotation significantly. Biochemical analysis suggests ortho ring cleavage is used throughout the aromatic metabolic pathways. We also recently demonstrated C. oleaginosus metabolizes monomers released from alkaline pretreated cornstover lignin, and the presence of lignin is not inhibitory to growth. With such an exceptionally desirable natural phenotype, this yeast could become a preferred host for oleochemical production if novel synthetic biology tools are developed. We identified both strong constitutive and phenolic-regulated promoters to drive expression of heterologous or native genes. In parallel, we are developing higher efficiency transformation methods for increasing the pace of engineering efforts. Overall, our work establishes C. oleaginosus as a promising platform to robustly convert all components of lignocellulosic biomass into novel high-value oleochemicals.

Metabolic engineering of non-model yeast Cutaneotrichosporon oleaginosus for valorization lignin and lignin-derived aromatics

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.